lattice engineering through nanoparticle–dna … · of biochemistry and cell biology, stony brook...
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1
Lattice Engineering via Nanoparticle-DNA Frameworks
Ye Tian1, Yugang Zhang1, Tong Wang2, Huolin L. Xin1, Huilin Li2,3, and Oleg Gang1*
1Center for Functional Nanomaterials, Brookhaven National Laboratory, Upton, NY 11973, USA. 2Biosciences Department, Brookhaven National Laboratory, Upton, New York 11973, USA. 3Department
of Biochemistry and Cell Biology, Stony Brook University, Stony Brook, New York 11794, USA.
Supplementary Information
Lattice engineering through nanoparticle–DNAframeworks
SUPPLEMENTARY INFORMATIONDOI: 10.1038/NMAT4571
NATURE MATERIALS | www.nature.com/naturematerials 1
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Part 1. Characterizations of polyhedral frames
Supplementary Figure 1. 1% agarose gel image under UV light for M13mp18 and DNA
frames. From left to right: Ladder (a), M13mp18 (b), octahedron (c), cube (d), elongated
square bipyramid (e), prism (f) and triangular bipyramid (g).
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Supplementary Figure 2. The size distribution of different frames, as marked, obtained
by Dynamic Light Scattering.
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Supplementary Figure 3. Negative stained TEM images of different polyhedral frames.
A. Octahedron; B. ESB; C. Cube; D. Prism.
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Supplementary Figure 4. 3D reconstruction of octahedral frame by cryo-EM technique.
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Supplementary Figure 5. Cryo-EM of Elongated Square Bipyramid (ESB) and 3D
Reconstruction. (A). Representative raw cryo-EM image of ESB structure with different
views highlighted by blue cycles. (B). Different models (top row) and relevant 2D
projections (bottom row) of reference-free class averages. (C). Surface-rendered 3D
density map of the ESB origami structure with different views. The density surface is
colored radially from interior red to outer green and blue. The values from the color key
mean the distance from the center in angstrom.
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Supplementary Figure 6. Representative cryo-EM image of cubic origami frame. Inset
is the representative reference-free 2D class average.
Supplementary Figure 7. Representative cryo-EM image of the prism frame structure.
Inset is the representative reference-free 2D class average.
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Part 2. Characterizations of nano-clusters by encoded gold NPs at the
corners of frames
Supplementary Figure 8. Representative negative stained TEM image of 10nm NPs
bound to octahedron frames at vertices, thus forming an octahedral nanocluster.
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Supplementary Figure 9. Representative negative stained TEM image of 10nm NPs
bound to ESB frames at the vertices, thus forming an ESB nanocluster. Cluster statistics
analysis reveals approximately 88% yield of 6 NP clusters (totally 150 clusters).
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Supplementary Figure 10. Representative negative stained TEM image of 10nm NPs
bound to cubic frames at the vertices, thus, forming a cubic nanocluster. Note, the cluster
is not skewed, unlike the cubic frame without particle (Supplementary Fig. 6). See main
text.
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Supplementary Figure 11. Representative negative stained TEM image of 10nm NP
NPs bound to prism frames at the vertices, thus forming a prism-shaped nanoclusters.
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Supplementary Figure 12. Representative agarose gel image (1%) under white light,
used for the purification of nanoclusters. The bands of products, free NPs and tracking
dye, are shown from top to bottom.
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Part 3. Discussions and Characterizations of frame/NP superlattices
In order to estimate the ratio between NPs and frames in the formed superlattices, we
measured the difference of the DNA concentration in origami solution before and after
assembly with NPs, since NPs can bind to the vertices of frames to form the aggregates.
Note, NPs could also bind to free staple DNAs in the solution; however, the melting
temperature for such binding is lower than room temperature. Let’s consider for example
an assembly of NP with octahedral frames. We mixed a certain amount (Vf) of octahedral
origami solution with different known concentrations of NPs, as shown in Supplementary
Figure 26 (10nm gold core for NPs, in 10mM PB buffer with 0.1M NaCl, denoted as
volume VNP and concentration CNP) that can bind to the origami frames, and annealed the
solution. At the certain NP concentration, the upper solution for the annealed samples
changes from clear to pink color, indicating the presence of unreacted particles. To avoid
the contribution from the DNA on particle, we characterized the samples upto the highest
NP concentration for which the solution remains clear after the reaction (and annealing)
of frames with NPs. In this case, we can find the highest possible amount of consumed
DNA for the full consumption of particles during the reaction with frames. For reacted
solution we estimated the amount of remaining DNA by measuring the OD value of the
upper clear solution of DNA at the wavelength of 260nm by UV-vis (OD1). Then we
prepared the control sample by mixing same amount of origami solution (Vf) and PBS
buffer (without NPs, VPBS=VNP) and measured the OD value (OD2) at the same
wavelength. Similar procedure is performed for all systems. Based on those
measurements we calculated the mole amount of origami frames and NPs individually in
the superlattice:
(1). Mole amount of origami=(OD2-OD1) × 50 ug/ml × (Vf + VNP) / (Nbase × 330
g/mole);
1OD(260nm)=50 ug/ml for double stranded DNA; Nbase represents the DNA base
numbers for the origami frame; mole weight for one DNA base is averaged as 330 g/mole.
(2). Mole amount of NP=VNP × CNP.
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From these experiments, we obtained the ratio of NPs to frames for different superlattices
assembled with the following frames: octahedron (NP: frame = 2.05:1), cube (1.87:1),
ESB (2.12:1), prism (2.28:1).
We should stress that the result of the ratios between NP and frames by UV-vis method is
a roughly estimation since some un-specific bindings of free DNA to gold nanoparticle or
frames may exist. Further experiments would be done to fine tune the models. For the
system assembled wit octahedral frames we performed the x-ray scattering studies to
verify the structure of the assembled framework at the different initial mixing ratios of
NPs and frames. Our study (Supplementary Figure 26) shows that the formed lattice
does not depend on the ratio; practically the same structure factor for all studied
conditions was observed.
SAXS experiments were carried out at the National Synchrotron Light Source’s (NSLS)
X-9 beamline and National Synchrotron Light Source II. The scattering data were
collected with a MAR CCD area detector and converted to 1D scattering intensity vs.
wave vector transfer, q = (4π/λ) sin (θ/2), where λ= 0.8551 Å and θ, are the wavelength
of incident X-ray and the scattering angle respectively. The scattering angle was
calibrated using silver behenate as a standard. The structure factor S(q) was calculated as
Ia(q)/Ip(q), where Ia(q) and Ip(q) are background corrected 1D scattering intensities
extracted by angular averaging of CCD images for assembled systems and dissociated
particles, respectively. The peak positions of S (q) were determined by fitting a
Lorentzian form. The structure factors were calculated using the SAXS-modeling
approaches previously reported in our studies 1-3
and discussed below. Briefly, the
structure factor S(q) is formulated by:
𝑆(𝑞) = 𝐶𝑍0(𝑞)
𝑃(𝑞)𝐺(𝑞) + [1 − (𝑞)𝐺(𝑞)] , where S(q) contains a structural term (lattice
peaks) 𝐶𝑍0(𝑞)
𝑃(𝑞)𝐺(𝑞) and a diffuse scattering term 1 − (𝑞)𝐺(𝑞).
In the diffuse scattering term, G(q) is a Debye-Waller thermal part: 𝐺(𝑞) = 𝑒−𝐷2𝑞2𝑎2
,
where a is the (smallest) lattice constant of the unit-cell, and σD, the Debye-Waller factor,
describes the relative variation of particle positions from their ideal lattice positions. Due
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to the size polydispersity, assuming a Gaussian distribution 𝑓(𝑟), of nanoparticles, this
position displacement effect smears the form factor P(q) and attenuates lattice peak
heights at the higher q-region. Such lost scattering intensity is accounted by the ratio
(𝑞) : (𝑞) =𝐹(𝑞) 2
𝐹(𝑞)2=
𝐹(𝑞) 2
P(q)=
∫ 𝐹(𝑞,𝑟)𝑓(𝑟)𝑑𝑟
∫ 𝑓(𝑟)𝑑𝑟2
∫ 𝐹(𝑞,𝑟)2
𝑓(𝑟)𝑑𝑟
∫ 𝑓(𝑟)𝑑𝑟
, where F(q) is particle’s form factor
amplitude.
In the structural term, C is a scaling factor, Z0(q) is a lattice factor for isotropic
distribution of grains, 𝑍0(𝑞) =1
𝑞2∑ 𝐹(𝑞ℎ𝑘𝑙) ∑ 𝑒2𝑖(𝑥𝑗ℎ+𝑦𝑗𝑘+𝑧𝑗𝑙)
𝑁𝑗
𝑗=1
2𝐿(𝑞 − 𝑞ℎ𝑘𝑙)𝑚ℎ𝑘𝑙
{ℎ𝑘𝑙} ,
where the inner sum is over the Nj particles in the unit cell, which have fractional
positions(𝑥𝑗 , 𝑦𝑗 , 𝑧𝑗); the outer sum is over the Miller indices (hkl) for the desired lattice
type. The function L is modeled with a normalized Lorentzian-peak shape. 𝐿(𝑞) =
/2
𝑞2+(/2)2 , where δ is the peak width. The correlation lengths () is estimated by =
2𝐾
,where K = ~0.9.
The main fitting parameters are lattice constant (a, b, c), peak width (δ), Debye-Waller
factor (σD), and the scaling factor (C). We used Python package ‘LMFIT’ as a curve
fitting engine. The fit results are plotted in Figure 3 and the main fitted parameters are
listed in Table-I.
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Supplementary Figure 13. Different binding modes of frames to NPs, examples for
cube (left) and prism (right) frames are shown. Red circles are 10nm gold NPs. Purple
lines are DNA linkers between NP and vertices of the frames. Blue squares and hexagons
represent the cubic and prism frames, respectively. The arrangement of polyhedral frames
around particles is chosen as observed for the corresponding lattices.
Calculation of unit cell parameters (a, b, c) for cubic frame system: dedge+2dshell.
We set the distance from the center of NP to corner of frames as d0 (defined by linking
DNA, in purple).
For cubic system (a=b=c): dshell= √3
3 d0.
a:c=1
For prism system (a=b): dshell= √15
5 d0 ;
(c): dshell= √5
5 d0 ;
a:c = (dedge+2 √15
5 d0) : (dedge+2
√5
5 d0) =0.87
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Supplementary Figure 14. Enlarged SAXS result (top) and a proposed model of FCC
lattice assembled from octahedral frames and NPs that satisfies the measured ratio of NPs
to frames (bottom).
(Å)
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Supplementary Figure 15. Enlarged SAXS result (top) and a proposed model of BCT
lattice assembled from ESB frames and NPs that satisfies the measured ratio of NPs to
frames (bottom).
(Å)
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Supplementary Figure 16. Enlarged SAXS result (top) and a proposed model of simple
cubic (SC) lattice assembled from cubic frames and NPs that satisfies the measured ratio
of NPs to frames (bottom). Red arrows point to two edge bundles between nearby NPs
that might experience the repulsion.
(Å)
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Supplementary Figure 17. Enlarged SAXS result (top) and a proposed model of simple
hexagonal lattice assembled from triangular prism frames and NPs that satisfies the
measured ratio of NPs to frames (bottom).
(Å)
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Supplementary Figure 18. Assembly of triangular bipyramid (TBP) frame with 10nm
gold nanoparticles. (A). Extracted structure factor S(q) v.s. q for TBP/NP assembly (black
curve with red points). Blue line is the fitting by dumbbell model dumbbell model 4,5
with
functional form S(q)~sin(dq)/dq, where d is the fit-obtained interparticle (center-to-center)
distance (58.4 nm); it matches well the designed parameter (56.3 nm). Inset is the model
of triangular bipyramid structure with 6HB as the edge. (B). SAXS 2D pattern for the
system. (C). Representative negative stained TEM image of TBP.
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Supplementary Figure 19. Representative negative stained TEM image of 10nm NP
bound to the vertices of triangular bipyramid (TBP) frames, thus forming TBP shaped
nanoclusters.
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Supplementary Figure 20. Probing melting temperature for cube-NP framework. The
sizes of aggregates are monitored as function temperature using DLS. The sample was
annealed from 53 ºC to room temperature at the rate of 3 ºC /h after assembly of NPs and
cubic frames. The determined melting temperature is around 35.9 ºC for the cube-NP
system. The melting temperatures for systems of other studied NP-DNA frameworks
were measured similarly, shown in Supplementary Table 2.
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Supplementary Figure 21. SAXS obtained structure factor S(q) for the system of
octahedral frames assembled with 20nm NPs, following a long annealing. The structure is
exhibits lower degree of order than the octahedra/10nmNP system (II). The lattice
parameter (a) for this structure (assuming FCC lattice) is 78.1nm, based on the position of
the first scattering peak. The distance matches well 10nmNP/octahedra system.
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Supplementary Figure 22. SAXS-obtained structure factor S(q) for the system of
octahedral frames assembled with 30nm NPs, measured after a long annealing. Compared
with 10nm and 20nm systems, the framework of 30nm NP does not exhibit structural
order.
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Supplementary Figure 23. Cryo-STEM of simple cubic system. (a) & (b) Low
magnification images of the lattice. (c) & (d) High magnification images of the (100)
plane (tilted).
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Supplementary Figure 24. Cryo-STEM of BCT system. (a) High magnification image
of (111) plane. (b) Low magnification image of the BCT lattice.
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Supplementary Figure 25. Cryo-STEM of FCC system (system III, as shown in Figure
2). (a) Low magnification image of the lattice. (b) High magnification image of (100)
plane of the lattice.
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Supplementary Figure 26. Structure factor S(q) for octahedral systems (system III as
shown in Figure 2) with different concentration of NPs and same concentration of M13
DNA (7.2nM). The result showed that more NPs would not change the structure (even
the orders of the structure) of formed superlattice.
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Supplementary Figure 27. Different possible models with high symmetry for
arrangements of cube frames that support a simple cubic structure of NPs lattice. The
measured ratio of NPs to frames is close to 2 (1.87), see the part 3 of supplementary
information. (a) the ratio of NPs to frames in the model is 1:1; (b) the ratio of NPs to
frames in the model is 2:1 (due to the significant edge-to-edge repulsion between DNA
frames, this model is less likely to be realized in comparison with the proposed
arrangement in Fig. 3 and Supplementary Fig. 16); (c) the ratio of NPs to frames for this
arrangement is 4:1.
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Supplementary Figure 28. Other possible models with high symmetry for the
arrangement of triangular prism frames in the simple hexagonal lattice formed by NPs, as
observed by SAXS. (a) the ratio of NPs to frames in this model is 1:2; (b) the ratio of NPs
to frames in the model is 1:1.
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Supplementary Figure 29. A possible model of superlattice that can support FCC
structure of NPs lattice formed with octahedral frames. Different from the one shown in
Figure 3, red marked voids are filled with another octahedral frame. Ratio of NPs and
frames in this case is 1:1. However, since the ratio of NPs and frames in the superlattice
is close to 2:1, this scenario is not realized.
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System
Number Linker
Motif Lattice ξ
(μm) I 0-6-10 Amorphous 0.300
II 2-6-10 FCC 0.632
III 9-6-10 FCC 0.431
IV 6-9-21 Amorphous 0.307
V 0-15-15 Amorphous 0.295
Supplementary Table 1. Correlation length (ξ) for NP-DNA framework systems formed
with octahedra frames, as obtained from the resolution-corrected first peak of the
measured structure factors.
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Polyhedron Binding method M.T. (oC)
Octahedron 15polyA/6bind thio-DNA (16mer) 35.5
ESD 8polyA/6bind thio-DNA (16mer) 28.8
ESD 15polyA/6bind thio-DNA (16mer) 32.4
Cubic 15polyA/9bind thio-DNA (30mer) 35.9
Prism 10random/8bind thio-DNA (50mer) 37.9
TBP 15random/6bind thio-DNA (16mer) 37.8
Supplementary Table 2. Measured melting temperature (M.T.) for all studied
framework systems with 10 nm NP. The measurements were conducted using DLS, as
described and shown in Supplementary Figure 20.
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Linker Motif
l-m-n
Sequences on TBP DNA sequences on NP
2-8-8 5’-TBP-TTATGAAGGT 5’-NP-TTTTTTTTACCTTCAT
2-8-16 5’-TBP-TTATGAAGGT 5’-NP-TTTTTTTTTTTTTTTT
ACCTTCAT
2-8-22 5’-TBP-TTATGAAGGT 5’-NP-TTTTTTTTTTTTTTTT
TTTTTTACCTTCAT
2-8-32 5’-TBP-TTATGAAGGT 5’-NP-TTTTTTTTTTTTTTTTT
TTTTTTTTTTTTTTTACCTTCAT
2-8-42 5’-TBP-TTATGAAGGT 5’-NP-TTTTTTTTTTTTTTTTT
TTTTTTTTTTTTTTTTTTTTTTTTT
ACCTTCAT
7-8-8 5’-TBP-TTTTTTTATGAAGGT 5’-NP-TTTTTTTTACCTTCAT
7-8-16 5’-TBP-TTTTTTTATGAAGGT 5’-NP-TTTTTTTTTTTTTTTT
ACCTTCAT
7-8-22 5’-TBP-TTTTTTTATGAAGGT 5’-NP-TTTTTTTTTTTTTTTT
TTTTTTACCTTCAT
7-8-32 5’-TBP-TTTTTTTATGAAGGT 5’-NP-TTTTTTTTTTTTTTTTT
TTTTTTTTTTTTTTTACCTTCAT
7-8-42 5’-TBP-TTTTTTTATGAAGGT 5’-NP-TTTTTTTTTTTTTTTTT
TTTTTTTTTTTTTTTTTTTTTTTTT
ACCTTCAT
Supplementary Table 3. The list of studied triangular bipyramid (TBP) frame systems
with different linker motifs used for the TBP/NP assemblies.
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3. DNA Sequence Design
(1). HPLC purified DNA oligonucleotides (attached on NPs) were purchased as
lyophilized powers (Integrated DNA Technologies Inc.). The staple DNA for origami
formations was purified with standard desalting method. M13mp18 single stranded phage
DNA was purchased from Bayou Biolabs, LLC and used without further purification.
The DNA sequences (5’ to 3’) were:
Name Used systems Sequence details
Seq. 1 System I-III for octahedron/ ESB HS-C6H12-TTCTCCACAATTTTTT
Seq. 2 System IV for octahedron HS-C6H12-
TTTTTTTTTTTTTTTTTTTTTTTTTTTT
Seq.3 System V for octahedron/cube HS-C6H12-
TTTTTTCTATCCTTACCACTATTTTTTTTT
Seq. 4 Prism HS-C6H12-TTTTTTTTTTTTTTTTTTTTTT
TTTTAACCTAACCTTCAT
(2). Staple sequences for octahedral system:
TCAAAGCGAACCAGACCGTTTTATATAGTC
GCTTTGAGGACTAAAGAGCAACGGGGAGTT
GTAAATCGTCGCTATTGAATAACTCAAGAA
AAGCCTTAAATCAAGACTTGCGGAGCAAAT
ATTTTAAGAACTGGCTTGAATTATCAGTGA
GTTAAAATTCGCATTATAAACGTAAACTAG
AGCACCATTACCATTACAGCAAATGACGGA
ATTGCGTAGATTTTCAAAACAGATTGTTTG
TAACCTGTTTAGCTATTTTCGCATTCATTC
GTCAGAGGGTAATTGAGAACACCAAAATA
CTCCAGCCAGCTTTCCCCTCAGGACGTTGG
GTCCACTATTAAAGAACCAGTTTTGGTTCC
TAAAGGTGGCAACATAGTAGAAAATAATAA
GATAAGTCCTGAACAACTGTTTAAAGAGAA
GGTAATAGTAAAATGTAAGTTTTACACTAT
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TCAGAACCGCCACCCTCTCAGAGTATTAGC
AAGGGAACCGAACTGAGCAGACGGTATCAT
GTAAAGATTCAAAAGGCCTGAGTTGACCCT
AGGCGTTAAATAAGAAGACCGTGTCGCAAG
CAGGTCGACTCTAGAGCAAGCTTCAAGGCG
CAGAGCCACCACCCTCTCAGAACTCGAGAG
TTCACGTTGAAAATCTTGCGAATGGGATTT
AAGTTTTAACGGGGTCGGAGTGTAGAATGG
TTGCGTATTGGGCGCCCGCGGGGTGCGCTC
GTCACCAGAGCCATGGTGAATTATCACCAATCAGAAAAGCCT
GGACAGAGTTACTTTGTCGAAATCCGCGTGTATCACCGTACG
CAACATGATTTACGAGCATGGAATAAGTAAGACGACAATAAA
AACCAGACGCTACGTTAATAAAACGAACATACCACATTCAGG
TGACCTACTAGAAAAAGCCCCAGGCAAAGCAATTTCATCTTC
TGCCGGAAGGGGACTCGTAACCGTGCATTATATTTTAGTTCT
AGAACCCCAAATCACCATCTGCGGAATCGAATAAAAATTTTT
GCTCCATTGTGTACCGTAACACTGAGTTAGTTAGCGTAACCT
AGTACCGAATAGGAACCCAAACGGTGTAACCTCAGGAGGTTT
CAGTTTGAATGTTTAGTATCATATGCGTAGAATCGCCATAGC
AAGATTGTTTTTTAACCAAGAAACCATCGACCCAAAAACAGG
TCAGAGCGCCACCACATAATCAAAATCAGAACGAGTAGTATG
GATGGTTGGGAAGAAAAATCCACCAGAAATAATTGGGCTTGA
CTCCTTAACGTAGAAACCAATCAATAATTCATCGAGAACAGA
AGACACCTTACGCAGAACTGGCATGATTTTCTGTCCAGACAA
GCCAGCTAGGCGATAGCTTAGATTAAGACCTTTTTAACCTGT
CCGACTTATTAGGAACGCCATCAAAAATGAGTAACAACCCCA
GTCCAATAGCGAGAACCAGACGACGATATTCAACGCAAGGGA
CCAAAATACAATATGATATTCAACCGTTAGGCTATCAGGTAA
AACAGTACTTGAAAACATATGAGACGGGTCTTTTTTAATGGA
TTTCACCGCATTAAAGTCGGGAAACCTGATTTGAATTACCCA
GAGAATAGAGCCTTACCGTCTATCAAATGGAGCGGAATTAGA
ATAATTAAATTTAAAAAACTTTTTCAAACTTTTAACAACGCC
GCACCCAGCGTTTTTTATCCGGTATTCTAGGCGAATTATTCA
GGAAGCGCCCACAAACAGTTAATGCCCCGACTCCTCAAGATA
GTTTGCCTATTCACAGGCAGGTCAGACGCCACCACACCACCC
CGCGAGCTTAGTTTTTCCCAATTCTGCGCAAGTGTAAAGCCT
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AGAAGCAACCAAGCCAAAAGAATACACTAATGCCAAAACTCC
ATTAAGTATAAAGCGGCAAGGCAAAGAAACTAATAGGGTACC
CAGTGCCTACATGGGAATTTACCGTTCCACAAGTAAGCAGAT
ATAAGGCGCCAAAAGTTGAGATTTAGGATAACGGACCAGTCA
TGCTAAACAGATGAAGAAACCACCAGAATTTAAAAAAAGGCT
CAGCCTTGGTTTTGTATTAAGAGGCTGACTGCCTATATCAGA
CGGAATAATTCAACCCAGCGCCAAAGACTTATTTTAACGCAA
CGCCTGAATTACCCTAATCTTGACAAGACAGACCATGAAAGA
ACGCGAGGCTACAACAGTACCTTTTACAAATCGCGCAGAGAA
CAGCGAACATTAAAAGAGAGTACCTTTACTGAATATAATGAA
GGACGTTTAATTTCGACGAGAAACACCACCACTAATGCAGAT
AAAGCGCCAAAGTTTATCTTACCGAAGCCCAATAATGAGTAA
GAGCTCGTTGTAAACGCCAGGGTTTTCCAAAGCAATAAAGCC
AATTATTGTTTTCATGCCTTTAGCGTCAGATAGCACGGAAAC
AAGTTTCAGACAGCCGGGATCGTCACCCTTCTGTAGCTCAAC
ACAAAGAAATTTAGGTAGGGCTTAATTGTATACAACGGAATC
AACAAAAATAACTAGGTCTGAGAGACTACGCTGAGTTTCCCT
CATAACCTAAATCAACAGTTCAGAAAACGTCATAAGGATAGC
CACGACGAATTCGTGTGGCATCAATTCTTTAGCAAAATTACG
CCTACCAACAGTAATTTTATCCTGAATCAAACAGCCATATGA
GATTATAAAGAAACGCCAGTTACAAAATTTACCAACGTCAGA
AGTAGATTGAAAAGAATCATGGTCATAGCCGGAAGCATAAGT
TAGAATCCATAAATCATTTAACAATTTCTCCCGGCTTAGGTT
AAAGGCCAAATATGTTAGAGCTTAATTGATTGCTCCATGAGG
CCAAAAGGAAAGGACAACAGTTTCAGCGAATCATCATATTCC
GAAATCGATAACCGGATACCGATAGTTGTATCAGCTCCAACG
TGAATATTATCAAAATAATGGAAGGGTTAATATTTATCCCAA
GAGGAAGCAGGATTCGGGTAAAATACGTAAAACACCCCCCAG
GGTTGATTTTCCAGCAGACAGCCCTCATTCGTCACGGGATAG
CAAGCCCCCACCCTTAGCCCGGAATAGGACGATCTAAAGTTT
TGTAGATATTACGCGGCGATCGGTGCGGGCGCCATCTTCTGG
CATCCTATTCAGCTAAAAGGTAAAGTAAAAAGCAAGCCGTTT
CAGCTCATATAAGCGTACCCCGGTTGATGTGTCGGATTCTCC
CATGTCACAAACGGCATTAAATGTGAGCAATTCGCGTTAAAT
AGCGTCACGTATAAGAATTGAGTTAAGCCCTTTTTAAGAAAG
TATAAAGCATCGTAACCAAGTACCGCACCGGCTGTAATATCC
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ATAGCCCGCGAAAATAATTGTATCGGTTCGCCGACAATGAGT
AGACAGTTCATATAGGAGAAGCCTTTATAACATTGCCTGAGA
AACAGGTCCCGAAATTGCATCAAAAAGATCTTTGATCATCAG
ACTGCCCTTGCCCCGTTGCAGCAAGCGGCAACAGCTTTTTCT
TCAAAGGGAGATAGCCCTTATAAATCAAGACAACAACCATCG
GTAATACGCAAACATGAGAGATCTACAACTAGCTGAGGCCGG
GAGATAACATTAGAAGAATAACATAAAAAGGAAGGATTAGGA
CAGATATTACCTGAATACCAAGTTACAATCGGGAGCTATTTT
CATATAACTAATGAACACAACATACGAGCTGTTTCTTTGGGG
ATGTTTTGCTTTTGATCGGAACGAGGGTACTTTTTCTTTTGATAAGAGGTCATT
GGGGTGCCAGTTGAGACCATTAGATACAATTTTCACTGTGTGAAATTGTTATCC
CTTCGCTGGGCGCAGACGACAGTATCGGGGCACCGTCGCCATTCAGGCTGCGCA
TCAGAGCTGGGTAAACGACGGCCAGTGCGATCCCCGTAGTAGCATTAACATCCA
TTAGCGGTACAGAGCGGGAGAATTAACTGCGCTAATTTCGGAACCTATTATTCT
GATATTCTAAATTGAGCCGGAACGAGGCCCAACTTGGCGCATAGGCTGGCTGAC
TGTCGTCATAAGTACAGAACCGCCACCCATTTTCACAGTACAAACTACAACGCC
CGATTATAAGCGGAGACTTCAAATATCGCGGAAGCCTACGAAGGCACCAACCTA
AACATGTACGCGAGTGGTTTGAAATACCTAAACACATTCTTACCAGTATAAAGC
GTCTGGATTTTGCGTTTTAAATGCAATGGTGAGAAATAAATTAATGCCGGAGAG
GCCTTGAATCTTTTCCGGAACCGCCTCCCAGAGCCCAGAGCCGCCGCCAGCATT
CGCTGGTGCTTTCCTGAATCGGCCAACGAGGGTGGTGATTGCCCTTCACCGCCT
TGATTATCAACTTTACAACTAAAGGAATCCAAAAAGTTTGAGTAACATTATCAT
ACATAACTTGCCCTAACTTTAATCATTGCATTATAACAACATTATTACAGGTAG
GTAGCGCCATTAAATTGGGAATTAGAGCGCAAGGCGCACCGTAATCAGTAGCGA
TTATTTTTACCGACAATGCAGAACGCGCGAAAAATCTTTCCTTATCATTCCAAG
TTTCAATAGAAGGCAGCGAACCTCCCGATTAGTTGAAACAATAACGGATTCGCC
GGGCGACCCCAAAAGTATGTTAGCAAACTAAAAGAGTCACAATCAATAGAAAAT
AGCCGAAAGTCTCTCTTTTGATGATACAAGTGCCTTAAGAGCAAGAAACAATGA
GTGGGAAATCATATAAATATTTAAATTGAATTTTTGTCTGGCCTTCCTGTAGCC
CCCACGCGCAAAATGGTTGAGTGTTGTTCGTGGACTTGCTTTCGAGGTGAATTT
ATGACCACTCGTTTGGCTTTTGCAAAAGTTAGACTATATTCATTGAATCCCCCT
TCCAAATCTTCTGAATTATTTGCACGTAGGTTTAACGCTAACGAGCGTCTTTCC
GGGTTATTTAATTACAATATATGTGAGTAATTAATAAGAGTCAATAGTGAATTT
Sticky end sequences for system I:
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40
TTTGCGGATGGCCAACTAAAGTACGGGCTTGCAGCTACAGAGAAAAAA
CTTCATCAAGAGAAATCAACGTAACAGAGATTTGTCAATCATAAAAAA
AAAGATTCATCAGGAATTACGAGGCATGCTCATCCTTATGCGAAAAAA
ATAAATCATACATAAATCGGTTGTACTGTGCTGGCATGCCTGAAAAAA
GGTAGCTATTTTAGAGAATCGATGAAAACATTAAATGTGTAGAAAAAA
CAAATGCTTTAAAAAATCAGGTCTTTAAGAGCAGCCAGAGGGAAAAAA
TCATATGGTTTACGATTGAGGGAGGGAAACGCAATACATACAAAAAAA
CAACGCTCAACAGCAGAGGCATTTTCAATCCAATGATAAATAAAAAAA
AATAGCAATAGCACCAGAAGGAAACCTAAAGCCACTGGTAATAAAAAA
GACAGGAGGTTGAAACAAATAAATCCGCCCCCTCCGCCACCCAAAAAA
AGCTTTCATCAACGGATTGACCGTAAAATCGTATAATATTTTAAAAAA
AGAGCCTAATTTGATTTTTTGTTTAAATCCTGAAATAAAGAAAAAAAA
GCTCACAATTCCGTGAGCTAACTCACTGGAAGTAATGGTCAAAAAAAA
CTTAAACAGCTTATATATTCGGTCGCTTGATGGGGAACAAGAAAAAAA
AAACGAAAGAGGGCGAAACAAAGTACTGACTATATTCGAGCTAAAAAA
ACTGTTGGGAAGCAGCTGGCGAAAGGATAGGTCAAGATCGCAAAAAAA
GGCCCTGAGAGAAGCAGGCGAAAATCATTGCGTAGAGGCGGTAAAAAA
AACGGGTATTAAGGAATCATTACCGCCAGTAATTCAACAATAAAAAAA
CAGAATCAAGTTTCGGCATTTTCGGTTAAATATATCACCAGTAAAAAA
GAAACATGAAAGCTCAGTACCAGGCGAAAAATGCTGAACAAAAAAAAA
ATCAAAATCATATATGTAAATGCTGAACAAACACTTGCTTCTAAAAAA
TGATTGCTTTGAGCAAAAGAAGATGAAATAGCAGAGGTTTTGAAAAAA
TTTGCGGAACAATGGCAATTCATCAATCTGTATAATAATTTTAAAAAA
TGTAGCATTCCAACGTTAGTAAATGAAGTGCCGCGCCACCCTAAAAAA
Sticky end sequences for system II:
TTTGCGGATGGCCAACTAAAGTACGGGCTTGCAGCTACAGAGAAAAAAAA
CTTCATCAAGAGAAATCAACGTAACAGAGATTTGTCAATCATAAAAAAAA
AAAGATTCATCAGGAATTACGAGGCATGCTCATCCTTATGCGAAAAAAAA
ATAAATCATACATAAATCGGTTGTACTGTGCTGGCATGCCTGAAAAAAAA
GGTAGCTATTTTAGAGAATCGATGAAAACATTAAATGTGTAGAAAAAAAA
CAAATGCTTTAAAAAATCAGGTCTTTAAGAGCAGCCAGAGGGAAAAAAAA
TCATATGGTTTACGATTGAGGGAGGGAAACGCAATACATACAAAAAAAAA
CAACGCTCAACAGCAGAGGCATTTTCAATCCAATGATAAATAAAAAAAAA
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41
AATAGCAATAGCACCAGAAGGAAACCTAAAGCCACTGGTAATAAAAAAAA
GACAGGAGGTTGAAACAAATAAATCCGCCCCCTCCGCCACCCAAAAAAAA
AGCTTTCATCAACGGATTGACCGTAAAATCGTATAATATTTTAAAAAAAA
AGAGCCTAATTTGATTTTTTGTTTAAATCCTGAAATAAAGAAAAAAAAAA
GCTCACAATTCCGTGAGCTAACTCACTGGAAGTAATGGTCAAAAAAAAAA
CTTAAACAGCTTATATATTCGGTCGCTTGATGGGGAACAAGAAAAAAAAA
AAACGAAAGAGGGCGAAACAAAGTACTGACTATATTCGAGCTAAAAAAAA
ACTGTTGGGAAGCAGCTGGCGAAAGGATAGGTCAAGATCGCAAAAAAAAA
GGCCCTGAGAGAAGCAGGCGAAAATCATTGCGTAGAGGCGGTAAAAAAAA
AACGGGTATTAAGGAATCATTACCGCCAGTAATTCAACAATAAAAAAAAA
CAGAATCAAGTTTCGGCATTTTCGGTTAAATATATCACCAGTAAAAAAAA
GAAACATGAAAGCTCAGTACCAGGCGAAAAATGCTGAACAAAAAAAAAAA
ATCAAAATCATATATGTAAATGCTGAACAAACACTTGCTTCTAAAAAAAA
TGATTGCTTTGAGCAAAAGAAGATGAAATAGCAGAGGTTTTGAAAAAAAA
TTTGCGGAACAATGGCAATTCATCAATCTGTATAATAATTTTAAAAAAAA
TGTAGCATTCCAACGTTAGTAAATGAAGTGCCGCGCCACCCTAAAAAAAA
Sticky end sequences for system III-V:
TTTGCGGATGGCCAACTAAAGTACGGGCTTGCAGCTACAGAGAAAAAAAAAAAAAAA
CTTCATCAAGAGAAATCAACGTAACAGAGATTTGTCAATCATAAAAAAAAAAAAAAA
AAAGATTCATCAGGAATTACGAGGCATGCTCATCCTTATGCGAAAAAAAAAAAAAAA
ATAAATCATACATAAATCGGTTGTACTGTGCTGGCATGCCTGAAAAAAAAAAAAAAA
GGTAGCTATTTTAGAGAATCGATGAAAACATTAAATGTGTAGAAAAAAAAAAAAAAA
CAAATGCTTTAAAAAATCAGGTCTTTAAGAGCAGCCAGAGGGAAAAAAAAAAAAAAA
TCATATGGTTTACGATTGAGGGAGGGAAACGCAATACATACAAAAAAAAAAAAAAAA
CAACGCTCAACAGCAGAGGCATTTTCAATCCAATGATAAATAAAAAAAAAAAAAAAA
AATAGCAATAGCACCAGAAGGAAACCTAAAGCCACTGGTAATAAAAAAAAAAAAAAA
GACAGGAGGTTGAAACAAATAAATCCGCCCCCTCCGCCACCCAAAAAAAAAAAAAAA
AGCTTTCATCAACGGATTGACCGTAAAATCGTATAATATTTTAAAAAAAAAAAAAAA
AGAGCCTAATTTGATTTTTTGTTTAAATCCTGAAATAAAGAAAAAAAAAAAAAAAAA
GCTCACAATTCCGTGAGCTAACTCACTGGAAGTAATGGTCAAAAAAAAAAAAAAAAA
CTTAAACAGCTTATATATTCGGTCGCTTGATGGGGAACAAGAAAAAAAAAAAAAAAA
AAACGAAAGAGGGCGAAACAAAGTACTGACTATATTCGAGCTAAAAAAAAAAAAAAA
ACTGTTGGGAAGCAGCTGGCGAAAGGATAGGTCAAGATCGCAAAAAAAAAAAAAAAA
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42
GGCCCTGAGAGAAGCAGGCGAAAATCATTGCGTAGAGGCGGTAAAAAAAAAAAAAAA
AACGGGTATTAAGGAATCATTACCGCCAGTAATTCAACAATAAAAAAAAAAAAAAAA
CAGAATCAAGTTTCGGCATTTTCGGTTAAATATATCACCAGTAAAAAAAAAAAAAAA
GAAACATGAAAGCTCAGTACCAGGCGAAAAATGCTGAACAAAAAAAAAAAAAAAAAA
ATCAAAATCATATATGTAAATGCTGAACAAACACTTGCTTCTAAAAAAAAAAAAAAA
TGATTGCTTTGAGCAAAAGAAGATGAAATAGCAGAGGTTTTGAAAAAAAAAAAAAAA
TTTGCGGAACAATGGCAATTCATCAATCTGTATAATAATTTTAAAAAAAAAAAAAAA
TGTAGCATTCCAACGTTAGTAAATGAAGTGCCGCGCCACCCTAAAAAAAAAAAAAAA
(3). Staple sequences for cubic system
TAAATATTGACGGAAAATTGAGGTTGTCAC
CCGAACAAAGTTACCAAAAAGTATAAGCCC
TGAATCGGCCAACGCGGTGCCAGAATGAGT
AATCGTAAAACTAGCAAGAATCGGGGTAGC
AGAAGCCTTTATTTCAGTAATACGCAAAAT
ATTGTGAATTACCTTAAATTTCATCAGTGA
ATAGAAAGGAACAACTTTTCAGCTAGCGTA
CATGTAATTTAGGCAGTATTTAATGCGTTA
ACGTTGTAAAACGACGGGTTTTCAAGGGCG
GACTTCAAATATCGCGAAGAGGAAATCAAA
CAAATAAGAAACGATTATTATTTGAATCTT
GAGTGAATAACCTTGCATAAATCTCAAGAA
ATTGCGTAGATTTTCAAAACAGATTGTTTG
GTCCACTATTAAAGAACCAGTTTTGGTTCC
CCCAATTCTGCGAACGCATATAAAATATAA
ATTTACCGTTCCAGTAAAAGCGCTTGAGGC
GGCTTTTGCAAAAGAAAACCAAACAAAAGG
AACTTTTTCAAATATAACAAAGATTTTAAC
CGTACTCAGGAGGTTTGGAATAGTCCTCAA
CCGTAATGGGATAGGTACAAACGAAAATAA
TAATCAAAATCACCGGGTTTGCCGTTTGCC
AGCCGTTTTTATTTTCTCATCGACCTAATT
AGCCGGAACGAGGCGCCTGCTCCCAAGCGC
ATCGGAACGAGGGTAGAGCAGCGAACCGAT
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43
AAAAATGAGTTACAGCGTCTTTCCAGAGAATCATCATATTCC
GAATTACATTCTAGAGGATCCCCGGGTAATCCGCTCACAATT
AAGCAAAGACATCTGCCAGTTTGAGGGGCCGCTTCTGGTGAG
ACCCCGGTGAGAGTCTACAAAGGCTATCTCGCAAGCGGTCCA
GAGAAGGGCCTGTACCATGTACCGTAACCCACCCTCCACCCT
TGAATATTATCAAAATAATGGAAGGGTTGCGCCTGTTTATCA
CCTACCAACAGTAATATAAAGTACCGACAATGCAGAACGCTC
GATTATAATAAGTCCAACATGTTCAGCTAAAAGGTCGTCAGA
CCTGATACCGAACTCACCGACTTGAGCCGGCCGGAAACGTCG
AAACGCACTTACCGGAAACAATGAAATATACACCATCAATAT
TACCGCGGTATTAAAACCAATCAATAATTCGCCTTAAATCAA
TAATAATTTGCTAATGTCGTCTTTCCAGATGCTTGATACCGA
GAAACAAAGCAGCAATTACCATTAGCAAATTTGGGCAATCAT
AGGTCAGAAACACTTACGAAGGCACCAAGGAAGTTTACATGG
CATTGCCTTGATAACCAGGGTGGTTTTTGAGAGAGTTGCAAC
TAAGCAAGAAACGCTAGCAAACGTAGAAGAACTGGGATAAAA
ATATTCGTCTGAAACCGTATAAACAGTTATAAGTTTACAGAG
TAAAGTAAGATACACAGTCAGGACGTTGGTAGAAAGATTCAC
AGAACGGCCCAATAGCAAGCCTCCCTCACACTTATCATTCCA
TGCTGTATACCACACAACATTATTACAGGGAAGAATTAGTTT
CAGAACCGGGTTGATTAGCGGGGTTTTGTACACCAGTACAAA
GAAATCGAATATCAAATTAACTGAACACAGAATAATCCAACG
CAGTAATGGGCTTAAGTATAAAGCCAACAGGCGAATTATTCA
ATCTTCTTGATGCAGGGTTATATAACTACTCAGTACCAGGCC
ATAAGGCCCAATAACTGAAAAGGTGGCAAATAACCTTAAGAA
AAGGGAAAATTGTGCGGAGATTTGTATCAGCACCAATGAAAC
TGGGATTTTTTTCACGTTGAAAATGTTTCCGAATTTTCTGTA
GAGTAGTATTATACTTTCGCAAATGGTCTCAATTCTACTAAC
TCAAAGCGCGGATTCCTGACTATTATAGTTCATCAACATTTA
GAGCTAAAGCTCATAACGTTAATATTTTAAAACAGAGGCGGT
TTCGCGTCCATTCGCCAGCTTTCCGGCAACGACGAGTGTAGA
AACAGTAAAGAGAACAGTACCTTTTACAAATCGCGCAGAGAA
CCGGAACGGTCATAGTAGCGCGTTTTCACGGCTGTCTTTCCC
GCTATTACTTTTTTCATTTAACAATTTCCAGCTGGCGAAAAA
TAACCCTGTAAAATCTCCAAAAAAAAGGATTTCTTAAACACA
CTGGCTCAAATTGGGACGAGAAACACCACAATAGTAGTAGCA
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44
TCGGTTGGAACCCTCGGAATACCCAAAAAATACATACATAAA
CTTTTGATCCTCATGCCTTGATATTCACTTGAGGCAAAAGAA
TAGCTATATAATAACATATATTTTAAATAGACAGTCAAATAA
TAAATGCGAACCGCCACCCTCAGAGCCAACTGAGTTTCGTTG
ACCAACGCAGATGAAGAAACCACCAGAATTTAAAATAACGTC
CCAAAGATCACCGTGACCAACTTTGAAACAAGAGTAATCTCG
TACGAGCTGCTATTCCTCCCGACTTGCGTTATCCGGAATCAT
CATTTTCCGCAAATCAGATATAGAAGGCGGAGGTTTTGAAGG
ATCGGTGCGGAAGCTGTGTGAAATTGTTCCGAGCTCCAAGCT
GGTAATACGTTTACGTAAGAGCAACACTACGTTAGTAAATCT
ATTTGCCAAAATAGACCGTCTATCAAATGGAGCGGAATTAGA
TTTAGCGCCACCAGACCCTCAGAGCCGCGAGCCGCGCCACCA
TATTTTTTTGCCCCCACCGCCTGGCCCTCTTTTCACATATGT
ATTTTTATACCAAATCAGAGCATAAAGCGCAAGGTGGCAACA
ACGATCTGCCGACATGCTTTCGAGGTGACTCCAAATTGCGAA
AATCAATATTACCCTGGCTGACCTTCATGAGGACATTAAAGG
AATAATAAACCGTTGTGAGAAAGGCCGGGCAATGCACCGAGG
TGGGCGCCCCGTCGTCCTGTAGCCAGCTTCCCGGAAACCAGG
AACAAAATTTATCAGACGCTGAGAAGAGCTTAGAAAATCGTC
TCAAAGGGAGATAGCCCTTATAAATCAACCCAGAGGGTAATT
TACAAATTACCTGAATACCAAGTTACAATCGGGAGTTCGAGC
AGGCCGCGGACTAAGGAGTGTACTGGTAAATGCCCCCTGCAC
AATTACGTTTAAACTATTCATTGAATCCAGACTGGCAGAGGG
GCAAGGCCTGCAGGTCGACTAATTTTCCTCGGGGGATGTGCT
GCTTTGATTTTGCGAGGCTTGCAGGGAGAACTATTTCGGAAC
GTCGAGAGCCACCCTCAGACCTAAATTTCACGGATAAGTGCC
TTGCGTATTTCCAGTAATTGCGTTGCGCAGATTAAATTTTTG
GTAACAAATCGTAACCGTGACCAGACCGGAAAATGTGAGCGA
AATCAGGATTTTTGTAATTGCTCCTTTTGAAGCAATCGAGCT
TGAATTACAAAAGGTCATATGGTTTACCATTGACAAGAACCG
CTCCGGCTAATTACTAAATAAGAATAAAAATGGTTTAATTTC
GACCATTCGGTGTCATGTTTTAAATATGGAATCAGTTGAGAT
AAATAAATGATACAAGACTTTTTCATGACCTAAAACGAAAAA
ATAGCCCGCGAAAACAGCCTTTACAGAGCCTGAACAAAGTTA
TGCCCGCTTGGGCGTCAGAAAAGCCCCAGTTAAAATTCGCGT
TGCATGCGATTAAGCTTCGCTATTACGCATTTCCACACAACA
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45
ACAATAGCTTCTGAATTATTTGCACGTAGGTTTAAAAAGTAATTCTGTCCAGAC
TAGTGAATTAATTAAATGGAAACAGTACTTCTGTATCCTTGAAAACATAGCGAT
AGAACCATCAGACTGCCCCCTTATTAGCAACCAGACACCCTCAGAACCGCCACC
AGAGGTCTCTTTACGCATCAAAAAGATTTTTTAATACTCCAACAGGTCAGGATT
TAGTTGCAAAGTTTACAACTTTCAACAGAAAGGAAAGGAGCCTTTAATTGTATC
GATATTCAGAGCAAAAGCCCTTTTTAAGGAAGGAACTGAGTAATGTGTAGGTAA
TGATTATCTAACGAAAATAAACAGCCATTTTTGTTGTTTGAGTAACATTATCAT
CAAATGCAGGCATACAGACGACGATAAAGTTTTGCATAGCGTCCAATACTGCGG
TTAAATCCTCACATTCGGGAAACCTGTCCGGGGAGGAAGATTGTATAAGCAAAT
TTAGGAAGCTCAACTGGAAGTTTCATTCAGTAGATAAATCTACGTTAATAAAAC
CTATTATGTCGCTGGGATCGTCACCCTCCAACGGCTTAACGGGGTCAGTGCCTT
CATCGATAGTACAATCGAAATCCGCGACAGACGGTAATTAGAGCCAGCAAAATC
GATATTCAGAAAATGCGACATTCAACCGTTATTCAGATGAACGGTGTACAGACC
TTAACATTTGCCCTGCTTGAGATGGTTTTGCGATTTGTTTAGCTATATTTTCAT
TTTCAATTCTTACCATTGAGAATCGCCAAGGCATTAAACAATAACGGATTCGCC
CAAAGCGCTGGCCTGATTCTCCGTGGGACACGTTGCAGTATCGGCCTCAGGAAG
GAGCGCTGCAAAATGGTTGAGTGTTGTTCGTGGACCATAAAAACAGGGAAGCGC
GGAATCATTAGGTTAATCCAATCGCAAGTTTTAGTTGAAATACCGACCGTGTGA
TATAAAATAAAGCCAACATTATGACCCTACGCAAGCATGATTAAGACTCCTTAT
CTACAACATTAGGATATAAGTATAGCCCAGTACCGCATTTTCAGGGATAGCAAG
GATTAGTATGTAGAACCAAGTACCGCACATCGTAGGTATTCTAAGAACGCGAGG
TACACTAACGATTGTAAAGCCAGAATGGAGCGTCATCCATTAAACGGGTAAAAT
TACGAGCCGGGCCTTTGGGTAACGCCAGGCCAGTGCGAATTCGTAATCATGGTC
CGCTGGTGAGAGATCTGGAGCAAACAAGTGTCAATCCAGTGAGACGGGCAACAG
ACCAGTAGCACCCCGTAATCAGTAGCATTATACATGTTACTTAAAAAAAAAAAAAAA
ATTAGACGGGAGGAGAGATAACCCACTTGATGGGGAACAAGAAAAAAAAAAAAAAAA
AGGCGCATAGGCAAATCAACGTAACAGTTTATTGAGGGAAGGAAAAAAAAAAAAAAA
CCCAATAGGAACGCATTCCACAGACACTGAGACGTGTATCACAAAAAAAAAAAAAAA
AGCTTAGATTAAAAATCATAGGTCTGACAAACAAATATATGTAAAAAAAAAAAAAAA
ACGTAATGCCACCATCTTTGACCCCCCAGGAGGAGTCTCTGAAAAAAAAAAAAAAAA
CTCAGAGCCACCAGCCGCCGCCAGCAGAATCAAATCTTTTCAAAAAAAAAAAAAAAAA
ATAGCTGTTTCCATAAAGTGTAAAGCTGTTGGGCCAGTCACGAAAAAAAAAAAAAAAA
GAGTAACAGTGCCATGAAAGTATTAACACGCATAAAGACAGCAAAAAAAAAAAAAAAA
AGATTCAAAAGGCTAGCTGATAAATTATTGAGTAGCAGATAGAAAAAAAAAAAAAAAA
GAACTAACGGAATTCAACTAATGCAGATTGCTGCAGTTGATTAAAAAAAAAAAAAAAA
TTTGCGGAACAATGGCAATTCATCAATTATCCTATCCCAATCAAAAAAAAAAAAAAAA
TACGCAGTATGTAAAGACACCACGGAAGAATTATTTTGCGGGAAAAAAAAAAAAAAAA
TGATTGCTTTGAGCAAAAGAAGATGATATCATACAACGCCAAAAAAAAAAAAAAAAAA
TTGGGGCGCGAGATCATACAGGCAAGTGCTCATACTTTAATCAAAAAAAAAAAAAAAA
CTGATTGCCCTTAGCAGGCGAAAATCCCGGAGAATGAACGGTAAAAAAAAAAAAAAAA
ATCGCACTCCAGCCATTCAGGCTGCGGCCATCAGCGGATTGAAAAAAAAAAAAAAAAA
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AATCGTCATAAAAGTTCAGAAAACGAATAACGCATAGCGAGAAAAAAAAAAAAAAAAA
CGTTTTAGCGAATTGCACCCAGCTACATCCCATGAACAAGCAAAAAAAAAAAAAAAAA
GACGACAATAAACTGAACAAGAAAAAATCCTGAAATAAAGAAAAAAAAAAAAAAAAAA
TAAATAAGGCGTTAGAAAAAGCCTGTACTACCTACGCGAGAAAAAAAAAAAAAAAAAA
GGTTTATCAGCTATGACAACAACCATTCATAGTGGAGTGAGAAAAAAAAAAAAAAAAA
ATTTAAATTGTATTTTTAACCAATAGGGTGCCTCTGCATTAAAAAAAAAAAAAAAAAA
AGAGAGTACCTTCGGATGGCTTAGAGGACCATAAGCCCGAAAAAAAAAAAAAAAAAAA
(4). Staple sequences for ESB system:
CCGACTTTGGGTTAATCGCAAGACAAAGTTAATTTTCAACCGATTGAGGGAGGG
TATCGGCCCAAAAAAAATCAGCTCATTTCGCGTCTAACGGCGGATTGACCGTAA
TTTAAATGCCGGAACGCAACTGTTGGGAGCCAGCTTGATAAGAGGTCATTTTTG
CATTATGTGATTCCGGTCAATAACCTGTAAAGGTGAAGGCAAAGAATTAGCAAA
CCTTGCTCAAGTTATGATGAAACAAACATTCATTTGTCTGTCCATCACGCAAAT
GAGTTAATTTGTCGAGAATAGAAAGGAAACGTTGACTTAAACAGCTTGATACCG
TACCTTTAGTAACAATTCCTGATTATCAGTTTGGACACGTAAAACAGAAATAAA
GCGACCTCGGAACGAGTTTCCATTAAACCAACCTAATTATACCAAGCGCGAAAC
CGAACCAACGCTCAGGCAGATTCACCAGCCAACAGTTTTGAATGGCTATTAGTC
CGCTGGTTTTCCTGTAATGAGTGAGCTACTTTCCAGCCAGGGTGGTTTTTCTTT
ATATGCGCAAACGTAAAGAAACGCAAAGAATAGAATGATAAATAAGGCGTTAAA
AGCCTAAAGCAAGCAAGAACGCGAGGCGTGAAGCCGCTACAATTTTATCCTGAA
TCGACAACTGCAACTGAACCTCAAATATAATCAACACTAATAGATTAGAGCCGT
GAATACCATAAGAAATTAGACGGGAGAAAATTGAGATAGCTATCTTACCGAAGC
ACGCCTGTGAGATTAGGCATAGTAAGAGCGATAAACTCAGAGCCACCACCCTCA
AGTTAATGCAAAATGGTTGAGTGTTGTTCGTGGACTGATACAGGAGTGTACTGG
ATTTATCGCGCCGCCGTTAGAATCAGAGTTTAGACTGTAAATCGTCGCTATTAA
TGGCAAGGCATTGATGATATTCACAAACCGCAGTCGACGGGGAAAGCCGGCGAA
CCATAAATAAGAGGGGCGGATAAGTGCCTAGGTGTTAGACTGGATAGCGTCCAA
AGAACAATTAATTGAAGTACCGACAAAAAACAACATAATTTACGAGCATGTAGA
TCGTAAATAAAAATAGATTCAAAAGGGTATATGATGAGATCTACAAAGGCTATC
TAATAAAGGGAACCGAGTAATCTTGACAACAAAGCAATTTCAACTTTAATCATT
CAGAGCCACCATTATAGCGACAGAATCATTCATCGAATCACCGGAACCAGAGCC
CTCTAGAGGATTGCTCAAATATCGCGTTAGCAAACGCCAGGGTTTTCCCAGTCA
CTGACCTATAAGGCTTGCCCTGACGAGAGGCGCATAGGCTGG
CTTTTTCAAAGAATACTCATCTTTGACCGCCTGATGAAATCC
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CAGAAGGAATAAGAGCAAGAAACAATGACCGAACAAAGTTAC
GCAAGGAACTAGCAGAGAGTCTGGAGCATTTTTGAATTCAAC
CAAAAGGATTAAAGGTGAAAAGGTGGCAACCAGCGTGGTTTG
GCCGCCATGTAGCGGGAAGGGAAGAAAGAGAGCTTTCTGAAT
TAATGTGGCTGATAAATTATGCTATTTTCCGCAATGCCTGAG
GGGGGATCAGGCTGACCAGGCAAAGCGCGAAGCTCAACATGT
ACACCGCCTCGTATCATTTGAGGATTTAACTAACAAGTTGAA
CGAGGAAAACGTCAAAAATGAAAATAGCTACAGAGCTAAAGA
ATCAGAGGAAGCGCACGATTTTTTGTTTACGCAATAATAACG
CTAAAGTAGGCCGCACAATGACAACAACTGAATTTAAATCTC
ATCATTTCGAAAGGAGCGGGAATAGCCCGCGAAAAAGCGTCA
TTCTACTCGCAAATCAATTCTGCGAACGTGTTGTAATCGGTA
TAACGTGAGAATCCGTGAGTGAATAACCACATAGCGATAGCT
TACATTTAATAGTACATCCAATAAATCAAAGCTAACCAAAAA
TGACCTAAAATCCATATAACTATATGTATATTATCACCGTCA
CATCAGTTAGCATTGCAAGCCCAATAGGCGCCACCAACCAAA
TAACAGTACCCTGTAGCCTCAGAGCATATACAGGCGCATCAA
GGAATTAAATGGAACTACCATATCAAAACGTCAGAGTAACAG
TTTGCTAAAAGCGTTTATTTTGTATCGGATACCATATGAAAT
AAATACCACTAGAAAAAGCTGCTGATGCAATTTAACCAAAGA
CGTTCTATAGGTAATTTTAGAACCCTCAAGGATGAACGGTAA
ACTTGCCATAATCAACAGTACATAAATCAGATTTCTATTCAC
AAGCATCGAGGAAGATATCTTTAGGAGCGAAGTATAAACAAT
AGGAATTACCTTGCAGTGCCACGCTGAGACTTTACTAGACGT
ATAGCGAAATTACGTAGGAATACCACATCAGTACAGTACCGT
GTAGGGCGCAAGCCATCGGCTGTCTTTCCCCATCCTGTTCAG
CCAGTCAGAGTAGTAAATTGGGCTTGAGACTGGCTCATTATA
ATCATAAACGAACTATGCGATTTTAAGAATGGTTTTGCTCAT
TCTGAATTCATCATTTATCATTTTGCGGTAATACATGAATGG
TCGGTCGAGTAAATGAATTTTCTGTATGGTCACCACGATAGC
GGATTATTGACCTGAATACGTGGCACAGAACATCGTACCGAA
TACATGGTTGAGTAACAGTGTCAGACGATCCAGTAACCGTCT
AATAGTATTGAATCCCCCTCAAATGCTTTTGCCAGAGTACCG
AATAAGAAGAACGCGCCTGTTTATCAACATTTTCGAGCCAGT
AACCGATTTTATCAGCTTGCTTTCGAGGCATCGCCCACGCAT
GCTTAATAAAATCATAGAATCCTTGAAATTGCTTCAGGAACG
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AAACAGGAGATAACCCACAAGAATTGAGAGAGAATAACATAA
AGAGGCAATGAGGAAGGGTAGCAACGGCAGGTGTCAAATTCC
AACGGGTCCTGAACAAGAAAAATAATATCTTATCATTCCAAG
CTTAGGTGAGCCATGACGGAAATTATTCGCGACATCATCTTC
GTACGCCCTTTCCTTACAGGGCGCGTACAGAGTCAATAGTGA
GAGCCGGTCGTAAGAAAGCGGCCAACGCTGATCGTGCTCAAG
TTAATTGATATAATGCTGTGGAAGCCCGATTAGAGAAGGCGA
CTGTAGCTTTTGTTCAGGAAGATTGTATGGGGACGACGACAG
TTATCCGGTATGCCGGAGAGGGTAGCTAAACAAGAGAATCGC
TTAAGTTCAAGCTTGCATGTTCGCCATTGTGCTGCAGTACCT
GGTCAGGAAAGACTATCAAAAAGATTAACACCTGCAGGTCGA
GCCCAATTTTGCCATAACGAGCGTCTTTGCACCCATTAAATC
AAAGTATTCAAAAAGTCATAAATATTCAAAATGTTATCACCG
CGGTCATTAATCAGGCAAGGCCGGAAACGGAACCGCTCAGAT
GTGCATCACAACCCGTCGGATTCTCCGTGGCGCATCGTAACC
ATAGCTGTTGCCCCCGGGCAACAGCTGAATTGGGCGTCGGGA
CTTTTTTAAGAAGACAAAATCGCGCAGAACTCAAATAACATC
CAGAGGCAAAGAACGGGTTTAGATAAGTATACCAGAAACCTA
ATGTTAGTTATACACCGGAATCATAATTGACCGTGAATTCAT
ACAGCATGCTCCATAGATTTGTATCATCCCCAGCGAAACGAA
CACCATTACCACCCGCCTCCCTCAGAGCTAATCAAGCATTTT
GCTGAGATATGGTTGCTTTAGTAGAAGAGGCGAATAATTACC
TGAATACGGTAATACAATACTTCTTTGATAAAAGAGAATTAC
TAGATTAAATATATTGAGAAGTGTTTTTTGGACGAGCACGTA
CAGTTCAGAGAAGGATTAGTTTCGTCACTCAACTAATAACGC
ACCCTTCTTACATTTGGAAATACCTACAATAAAAACCATTAC
AAGTTTGTACATCGATTTTCAGGTTTAATTATTTGTTATACT
AACACTGCAGAACCTTGCAAAAGAAGTTTAGATACATGCAAA
AGAGTTGCCGCTCACAATTCCACACAACTTTTGACCTGAAAT
AGCACCGAGCCCCCTTGCCATCTTTTCACGCCACCCCACCCT
TGAGCAAAATGGAAGTGAGGCCACCGAGTTAGTAACTATCGG
GAAATCGGCCCCCTACGGGGTCAGTGCCCTTTTGATCCAACG
TCAAAGGGAGATAGCCCTTATAAATCAAAGGCCCGTATAAAC
AACCAAGTAACAACGCCAACATGTAATTAACAAAGAAGGAGC
AACCTGTGGGTGCCTGTGAAATTGTTATCAGCAAGCGGTCCA
ATGGTTTACATATAAGAAAATACATACAAACTGTTTAGTATC
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AGTTTGAAAGCAAATATTTAAATTGTAAAGCCAGCAAATCTA
AAGATTAGTATTCTAAATCAGATATAGATATATTTTAAATAG
GTTTCAGAAGGCTCCAAAAGGAGCCTTTAACAACTTTCAACA
ATTAAATCAGCTTTCATCAACATTAAATTTGTTAAAATTCGC
CTGGCCCGCGGGGAGAGGCGGTTTGCGTTTGCCCTTCACCGC
CAAAAAACGGAGTGTCTTTCCAGACGTTCTGAGGCTTGCAGG
AATCCAACAAAAGAAAGTAAGCAGATAGAATAGCACGCTAAT
CAAAAGGGAGGCTTGCCACCCTCAGAACAACCCATAACTACA
TAATATTGTCTAAAGTTATGAGCGAGTATGATGAAAGCAACC
CAGACCAAAATTAAGTAGCCACCAGAACGGTTGACTTAGTAC
AGGAAAACCAGCAGACTGATAGCCCTAAACAATATAGATAGA
TCAGTGATCATCAAGAACTGACCAACTTAGAAAAATCTACGT
GTTGGGATGAAAGAGGACAGATGAACGGAGTAGATCATTAGA
TACTCAGAGTACCACTGAGACTCCTCAAGAAAACGAGAATGA
CTAATGCGAATATAAGAATCGCCATATTTACCGCACTCATCG
TTTACAGTTAAAACACACTAAGCCCAATAAGAGGAGCTTTAC
TTACCGTTTGGCCTCAGGAGGTTGAGGCAAGCGCTAGGGCGC
CCACCCTAGGATTAGCGGGGTTTTGCTCGAGGTTTAGGGGGT
TTCTGGTATGCAACAGCTTAATTGCTGACTCCTTTGGCGAAA
AAAAGCCCTCAGGACGTTGGTGTAGATGGGGAACAGGCCTTC
AAGCCTGCGTGCCAGCTGCATTAATGAAAAGCATAAAGTGTA
GCAAAGCGGATCCCACGACGGCCAGTGCGGGTAACTCCAACA
CAATATAATCCTGATTGATGATGATTTTAA
AATTACATTTAACAATTCAAGAAATTGCTT
TTGCGCTCACTGCCCGACTCACACATGGTC
AAGCCAGAATGGAAAGAAATAAACAGAGCC
GTCCACTATTAAAGAACCAGTTTTGGTTCC
CAGACTGTAGCGCGTTAGTTTGCCCAGTAG
TTGGGGCGCGAGCTGATTAGCTATTCCATA
GAATAATAATTTTTTCCAACTAATAACGAT
GCCTCTTCGCTATTACAGGGCGAGCACCGC
TTATTTTGTCACAATCACACCACACGCAGT
AGCGAACCAGACCGGATTAATTCGTCAGAA
CTCGTTTACCAGACGACAACACTAAAGATT
GACTTGCGGGAGGTTTTTTTAGCTTACCGC
AAAAGGGACATTCTGGTCACACGTTGCAAC
GGCCGATTAAAGGGATCGGGAGCCCGCCGC
AGTCAAATCACCATCAGAGAAAGTTTCAAC
GCCATCAAAAATAATTTTTAACCTAATCAG
ATCTGGTCAGTTGGCACAAACCCAGTATTA
CCAGACGACGACAATAGGTAAAGCTCAACA
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50
GCCACTACGAAGGCACGGGTAAAGCGAAAG
TTCAAATATATTTTAGAACGCGACCTCCGG
TAAGTATAGCCCGGAAGTCGAGAAAACATG
AACAAAGTCAGAGGGTTTAACTGTTATCCC
TACCCAAATCAACGTAAGAACCGACGGTCA
TTAATTTTCCCTTAGGTCTGAGAGACACCACACTAAACAGGAAAAAAAAAAAAAAAA
TCACCAGTGAGAAGCAGGCGAAAATCTCGTAATTTAATTGCGAAAAAAAAAAAAAAA
CGTGGCGAGAAAGTCACGCTGCGCGTCCACCACTCCTCATTAAAAAAAAAAAAAAAA
GAAATTGCGTAGGGAGAAACAATAACGTTATTAGCAATTCATAAAAAAAAAAAAAAA
AGGTCATTGCCTTGTCAATCATATGTGCCTTTAGCCGGAGACAAAAAAAAAAAAAAA
AACCAATCAATAGTTTTTATTTTCATGCCAACGTAATTCTGTAAAAAAAAAAAAAAA
TAAGAATAAACAAATTCTTACCAGTACCTTATTGGAATAAGTAAAAAAAAAAAAAAA
TTTTCAGGGATACCACAGACAGCCCTCAGGTAGATCATAACCAAAAAAAAAAAAAAA
AAAGTACAACGGGTTACTTAGCCGGACTCAGCAATACGTAATAAAAAAAAAAAAAAA
CAATAGATAATATAAATCCTTTGCCCGGCGGTCTCAATCAATAAAAAAAAAAAAAAA
ATTAAGCAATAAAATACTTTTGCGGGAGTTTCATATTTTCATAAAAAAAAAAAAAAA
TACTGCGGAATCTCAGGTCTTTACCCTATTCTGGGGTTGATAAAAAAAAAAAAAAAA
TCTTACCAACGCGTTACAAAATAAACGGAATCAGAACCTCCCAAAAAAAAAAAAAAA
TTTAATGCGCGAAAGATAAAACAGAGCCAGCCAACCAGTAATAAAAAAAAAAAAAAA
ATAGTTGCGCCGTTTTGCGGGATCGTGTTAGCGAGGAATTGCAAAAAAAAAAAAAAA
CCTTTTTAAGAAACTGGCATGATTAAATATTATAACACCCTGAAAAAAAAAAAAAAA
TGGGATAGGTCAAGATCGCACTCCAGCGGTTGAAATAGGAACAAAAAAAAAAAAAAA
TAATAAGTTTTAGCCTATTTCGGAACTTGATGGGGAACAAGAAAAAAAAAAAAAAAA
TAACCGTTGTAGTCCAGAACAATATTTCGCCTGAACAAAATTAAAAAAAAAAAAAAA
CGACGTTGTAAACGGGTACCGAGCTCTATTATAGAGCTTCAAAAAAAAAAAAAAAAA
ACCACCGGAACCTCAGAGCCGCCACCAAAATCACTTTAGCGTAAAAAAAAAAAAAAA
GTGAATTACCTTAACGGAACAACATTGGCGCAGGATATTCATAAAAAAAAAAAAAAA
CGGATGGCTTAGTAAAGTACGGTGTCCTTTCCGTCGGTGCGGAAAAAAAAAAAAAAA
AAGGTAAATATTTTGGGAATTAGAGCTTTTTAAGAAAACTTTAAAAAAAAAAAAAAA
(5). Staple sequences for prism system:
GCGCGTTTTCATCGGCGTTTGCCGAATTAG
CCGTTCTAGCTGATAAAATCACCATTTCAA
GCCATTAAAAATACCGGCGAACTCAGTAAT
TTTTGCTAAACAACTTAGTAAATCGTAACA
TTTGACCATTAGATACAATTCTGGATGGCT
TAACGGAATACCCAAAAAGGAAAATCAGAG
CGCCAGAATCCTGAGAAAGGGATGCGCGTA
GATTATCAGATGATGGGAGCGGATTGAGGA
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GAGGCATTTTCGAGCCCAACGCCTCATAAT
CATTAAACGGGTAAAAAAGACTTGCTGAGG
CAAGATTAGTTGCTATGCGGGAGTTATTTT
AACAAACGGCGGATTGACAACCCAATTTTT
CGTCATAAATATTCATGGATAGCGCAACAC
TCCTTGAAAACATAGCCTATTAAATGATGA
AGGCTTGCCCTGACGAACAAAGCCTTTGAA
TTGTTCCAGTTTGGAAAATAGCCCTGGCCC
TGTTTCCTGTGTGAAACGAATTCTAAGTTG
CCTGCCTATTTCGGAATGCCCGTAAAGCGC
TAGGATTCAGAACCGAACCGCCACCCTCGGCGCATACAAGAA
TGCAACATTGAAAGAGGTTATCTAAAATTGCGTTAAACAGTA
AAACCAAAACAACAAGGTGAGGCGGTCATGGCACATCTGACC
CAGTGAGAGCTAAACGCGCTTAATGCGCCTCAATCCATTGCA
AGCCAGCGAGACGGCGTGCCAGCTGCATCACATTAGTCATAG
AGAAGATATGGCTAGGCCAACAGAGATAAAGAACGCATGTAG
AATAATTGGGAAGCGACCGCTTTTGCAATGACCATGCATCTG
CCACCCTAGCGGGGCTTGAGATGGTTTAAATCTTGAGGCTGG
ACTCTAGACACAACCCACCGGAACCGCCGAACCACGCCTTGA
ACATGAAGGTCAGTCCGTTCCAGTAAGCATTCAACTATTACA
AGAGGACAGGGATAATCACCGTACTCAGAGGCGGACAGAACG
CCTGGGGGGGAGAGTATTGGGCGCCAGGACCATTAGATAGCA
ACAGGAACTTCTTTAGTAAAAGAGTCTGCCTCGTTGGGCGCG
TTTAGTTTTTAACCGTCAATAGTGAATTTCAATATACAATTT
GTTTCATCTGTTTAGGTGGCATCAATTCAAGCAATACCAAAA
ATAAAGTATTGAGATGTTTAGTATCATAATCTTTATGGCAAA
AGGTGGCACCGATTAGGTAAATATTGACCAATAATAGAAAAG
TTCAGGCGAGGGGATCAGAAAACGAGAAAAGAAGTCCAAAAT
CGGAAACGCTTGCGGCGGTCAAGCTTGCCATAAAGAAATCAC
GCAACGGCACCAACACACTCATCTTTGATTGTGTCGGAACGA
TATCATAGCTGGCGCGGAAACCAGGCAAGCCTCAGCCCTCAA
TTAAATATTTGCCCAATTTTAAAAGTTTAATAATGGAAGGGG
TAAATCCTGATTGCGCCTGCAACTAAAGTGGTTAGAACCTAC
TGAAAGCAGTGTCTTCGGCTATAAAGCCTCTGTCCCACCGCC
TAAAGAAAAATCCCCAAGCGGTCCACGCGGGAGAAGCCTTTA
CCGGATAAATTGGGTTTTGCTCAGTACCGAGGTTTCCACCCT
TCTTACCGTGAGGAGAATTAAGAATACGGTATTAAAGACGAC
TGAGAGATGAACAAACGTCAAAAATGAAAACAGCCCCACTAT
AATCCTGGAACAAAACTTTACAAACAATTACAAAAGGGAGAA
ATATAGAAACGTGGGCGAATCCTGTTTGGCGAAAAGTCTTTC
GGGCTTAACCGACAGCTGAGAGCCAGCAGGTCAGTGGAGCAC
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AGCGAGACGGGGAACTGTTCGTCTGGCCTAACCGTAAATCAA
ATCAATGAAAGCCGCGAGAAAGGAAGGGAGCAAGCAAGAACG
CAGAGAGTAATTTGTCCAACGTCAAAGGATGGTGGCCCCAGC
TTTTGCGATTGTTTTTGCACGTAAAACATTACATCTCGCGCA
CAGAGCACTGAAAAGCTATATTTTCATTTACGGTGACATGTT
TTGATGATCTGAGATCAGTATCAAGAGTATTTCAAGAAGGAT
CGCAAGGTAAACGTTGTACCCCGGTTGAAAACAAGCCGGAGA
TGAATAAGAGAAGATCCGGCTTAGGTTGTTTTTCACCGACCG
TACTATGTGAAATACATGGAGTAATGTGGGATTAACCGTTGT
ACAGCTTTAATAATAAGGAACAACTAAAGTTAGCGTAGCATT
CAGCAGCATTACCTAAACAGTACATAAATATCAAAATCATAA
CTAAAGTGAATAGATTTTTCACGTTGAAGGTGAATGACAATG
AAATCAGCGAACCAAAACTCCAACAGGTGGAACGCAGCCAGC
CAGAGCCAATAACAGAAGCGCATTAGACTGGTTTGTTCCGAA
CAATTCCAGGATCCTTTCCCAGTCACGAACGATTGCACCAGA
ACAATAAAAATTATGGATTATACTTCTGGAGTAACCTCGTAT
TAGAGCTGACCCTGGAATTAGCAAAATTTACTAATAAATGGT
AGTAGTATTCATTACGGTGTACAGACCAAGAGCCAAGTACCG
TTGCGAAGATAGCTAGCAATACCGATAGACGCCTGTAACGAT
CATCGTAGCTAGGGCCTAAAGGGAGCCCCCAAATCCCAGCTA
CAATGCCGTTGCTTGTATAACGTGCTTTTCCATCACGCAACT
GTTAAATATTGCTCACTTCAAATATCGCGTCAGAATGGGATA
GGCCAGTGTGGAGGTGACACACCAATGAATAATCATGTAAAG
GCACCGTATTAGCGTGAGTGAGCTAACTTAATGAATTCTTTT
CGAGGCGATCCTGAACGTGAACCATCACCCGATTTCGAAAGG
TTAACGGAGTATTATTATGCGATTTTAAACAACATTAATGCA
CTGAGTTCCATCGCCAGCTTGCTTTCGAAATCTCCTTTCAGC
GGGTAGCGAGAAAGTTTTTAGAACCCTCTATAAGCAAAAGCC
ATCAGGTCAAGCACTGACGGGAAGATTGATATATTAGATTCA
TAAGCAGCCTTATTAAAAGAAACGCAAAAAGACAAATTCATT
CATTTGAGATGACCTCTTCAAGACAGCAGCAGGTCTGAGAGA
GACAATATCAATAATTCCTTATCATTCCGAACCCTGACAATA
CATATCACGGATTCTTTGAATACCAAGTTCGACAAATTATCA
CCACAGAATGAAATATCTTACCGAAGCCCAAACGTAGAAAAA
TCAACAGGTGCCACAAAGGTAAAGTAATAACGCTCTACAAAT
AAGAATAGCAATGGTTTTTTCCATGTTAGGGATCGGAGGGTA
ATCGGCACGTGGACCCAGTTACAAAATAAATAGCATTAACTG
AGTCTCTAACGCCAAACGAACTAACGGAGAACTGGTTCTGAA
AGCAATAAAACGCTCCTACATTTTGACGCGCTACAAGAATCA
GACGGGGGCCCACTATCTTACCAACGCTGTATTCTAAATCAG
GGTCACGAACATTATTTTTTAACCAATACAGGATTTTCGAGC
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TTTCATCTTGGTGTCCCTGACTATTATAGTTTTAAAGAGAGT
ACGCGCGTGCCTAATTTGCCATCTTTTCAACCATCGCAAGGC
GCCGCCGAGAGCCAATACGAGCCGGAAGATGCCTGAAACGAC
AGGCGAAAGACAGGTAAAAGCTTATCCGAACGAGCACCGTCT
GGTAACGAAACAAACAGAGCCGCCACCATCCCTCACCGCTCA
AAAGGGTTATTTTTTGAGAGTCTGGAGCTAATCAGAAATATT
CCAGTTTTGCGCAAGGGCGATCGGTGCGATAAAAATTTGCCA
TTCAAAGGTCTTTAAGATGGGCGCATCGTTCCTGTCATCAAA
AGATAACAATTATCGTTTACCAGCGCCAGACACCAGCATGAT
GCGCGAGTATTATTGAACGAAGCTAAATTAGCTCATCTGGAA
ACCACCAATTTACACAGAACAATATTACTCACTTGTTATAAT
ATGCTTTTAATAGTTTTACCAGACGACGGGCCTCTTTCGCCA
AAAAGGGTACCGCATATCCCATCCTAATAGAACGCCACCAGC
CCAAAAACATTGCCGAGAGATCTACAAATAGGTAATTAAATG
TTTAGAAATTATTCAGTAACAGTACCTTGAAATAATCAATAT
TAAGACTATAGCCGAATTGAGTTAAGCCGGAAATTAAGGGCG
GGAGTGATTTGTCGCAGTACAAACTACATTGCGCCTTCTTAA
GGTAGAAAATTACCAGAGGCTGAGACTCACTGGTAATGGCTT
TACTAGATAATAGACTCAATCAATATCTGCAAATGAGAGAAT
GAGCGGGGCCACCGGATTAGTAATAACACGCCAGCGTCTGAA
ACGAAGGCTACAGAAAGGCCGCTTTTGCCTTAGCCGAAATCC
CTTGCAGACGGTCACATCGCCTGATAAACCCCCAGTGCCACT
CTGACCTTACCTCAGCCACCAGGAGTGTCTCAAGACTTTAAT
CATTGTGAGATTCATTTAGGAATACCACGTCATACATAAGTT
CCCCCTTAATCAGTCCAGTAGCACCATTGTGGTTTTCGGCCA
AGAAACACAAGGGAGAGGGGCCCTCATAGGATACATACATAA
GCGACCTCACTAAACTAAAACGAAAGAGTCGGAACTCACCCT
CTACCTTAATTTCATAAATTTAATGGTTACATTTAATGTGAG
TTTTTGAAAAACAGTGTTCAGCTAATGCTTACGAGGGTATTA
ACATTCAAACATATACGCAGTATGTTAGCTTTTTAAAGAGCA
CGGAACCCCAGCATTTGAGGCAGGTCAGCGTTGTACAGGTCG
AACAAACAATAAGGGAACGCGAGAAAACGGTTATATAGATTA
GAGGGGGAAACAGTCGACGACAGTATCGAGCGCCATCGCTAT
AGACGCTCCTTGCTAAACAAAATTAATTTGAAATAAATATAT
CAATTTTTTTTAGCTTACCGCGCCCAATAAGAAAGAGAGCTT
CAATAACTCCATATTGAATATAATGCTGCGGTTGTAAAGCCT
AGCGGGCGGAATCAGAACCTCCCGACTTTTTGCACAAGTTTTTTGGGGTCGAGG
GAGGCGAGTATTAGGAAACCACCAGAAGCAATTCAAGAAATTGCGTAGATTTTC
AACACCCGTTGCAGTTATAAATCAAAAGCAAGAGTATATTATTTATCCCAATCC
GATACATGAATTTAGCCTTGAGTAACAGCCTATTACTCATTATACCAGTCAGGA
ACAACAATCGTCACTCTTTCCAGACGTTTCAACAGAAAAAAAAGGCTCCAAAAG
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AAAGGTGCCACAAGAACAAAGTTACCAGAGAACTGCGGAATAAGTTTATTTTGT
ACCTTTACAGCTCAAATGTGAGCGAGTAACCGTAAGCAAAGCGGATTGCATCAA
AACCAAGACATTCTTTAGTCTTTAATGCAACGAACGCCTGTTTATCAACAATAG
TGTGATAATCAAGATCTGTAAATCGTCGGATAGCTTAACTATATGTAAATGCTG
CATTTTCAGATGAACCCAAATCAACGTAGAAACACTAAGTGCCGTCGAGAGGGT
ATGGATTCACCCGCCAGGAGGCCGATTAAGTGTTTCCTGAGTAGAAGAACTCAA
TACGCCAACCCTCGAAAATGTTTAGACTTGAATCCGAAGATCGCACTCCAGCCA
TATTCACCCAGGGTCCGGGTACCGAGCTTTGTTATGAGCCGCCACCCTCAGAAC
ACATTATTAATTGCAACAGTTGATTCCCATTTCGCAGTAGTAGCATTAACATCC
CACCAGTAAAATCAAGCGACAGAATCAAATTTTCGATTGCGTTGCGCTCACTGC
TAAATTGATAAAAAGCCGGAGACAGTCAATTAATGAGAATCGATGAACGGTAAT
GGCGCAGGGAGTTAGGCTTTGAGGACTATACGTAACGATTATACCAAGCGCGAA
TAACAACAAAAGCCATCGCCATATTTAAAGTAATAAAAAATCTAAAGCATCACC
AGGCAAGGCAAATAATACTTTTGCGGTTTTGCGCGAACGAGTACTCATGAAGGT
CAAGAAAAATAACTCATCGAGAACAAACACGACGATAGCCCTACTCATGAAGGT
CTAAATCGGAACCGCTGGCAAGTGTAGCCGTTTGTTTTGAAGACTCATGAAGGT
AAAATTCATATGACCGTCACCGACTTCGCTAATCCGAGGAAAACTCATGAAGGT
ATTTTTTGTTTAAGTCAGAGGGTAATTCACCGCCGAGATAGGACTCATGAAGGT
GGAGATTTGTATATCATAAGGGAACCTTCGGTCTTTCATGAGACTCATGAAGGT
GCTGGTAATATCTTGGCAGATTCACCTCACGCTTTTAGACAGACTCATGAAGGT
CGGGAAACCTGTGCAACAGCTGATTGCATTTGGTTTAGCGTCACTCATGAAGGT
TCGCAAGACAAACGTTAAATAAGAATAAAGAAGTTAATTTTCACTCATGAAGGT
GTATCGGTTTATCCACGCATAACCGACATGTACGAATTTTCTACTCATGAAGGT
CGCTTCTGGTGCAAAGGGGGATGTGCAGTAAGAGTCCAATACACTCATGAAGGT
AAATATCAAACCTTAGAGCCGTCAATACCGGAAAACATGTAAACTCATGAAGGT
CGGAATAGGTGTGCAAGCCCAATAGGTGACCAATGCTCATTCACTCATGAAGGT
GAAGCCCGAAAGCTTTTGATAAGAGGCGCATTAGTCGGATTCACTCATGAAGGT
CTACGTTAATAAAAAGGAATTACGAGAGAATGGATAAACAGTACTCATGAAGGT
ATGTCAATCATATAATATTTTGTTAAAGCCTTTATCAATATGACTCATGAAGGT
AGCCACCACCCTTAAATCCTCATTAAAGGCGATGTAATCATGACTCATGAAGGT
AGATGAATATACATTTCAATTACCTGAATACATATTATCATCACTCATGAAGGT
(6). Staple sequences for TBP system:
TTGTTCCAGTTTGGAAGAGATAGGAGAGAG
AGAGGCAAAAGAATACACCAACCCAGCATC
CCCAATCCAAATAAGACAGCCATCTTGCGG
CAGTCAAATCACCATCTGAGAAATACTTTT
AACAATTCGACAACTCAGTATTAGCAAATC
ACTTTAATCATTGTGAAGATGGTTGACAAG
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GTAGATTTAGTTTGACCCAATTCCGGATGG
CAGTTCAGAAAACGAGCCTCAAATAGCGAG
CCATCTTTTCATAATCCCCTTATCCATTAG
GTATTAAGAGGCTGAGATTCTGACTTTTGA
AACAATAGATAAGTCCAACGCGCTTAGGCA
AACAAACGGCGGATTGTCGGATTAAATTCG
CAACATATAAAAGAAAACATACAACAAAGT
TGAAAGCGTAAGAATATAGAACCATGGAAA
TCGCAAGACAAAGAACGCTGATGTAGATTA
TGAGAATAGAAAGGAAAACAGTTAGTACAA
CAAAATCGCGCAGAGGTTTGAATTAAAACA
GTGAAATTGTTATCCGCATAGCTATTAAGT
GGAGAGGTCACATTGAAGCATAAAGTGTTAGAGGATAAAACG
AAAAATCACATAACATTACGAGGCATAGGATAAAATGCCTGA
GAGCCTAAAAAATGACGGGAGAATTAACAACCCACGCTATCT
AGGTTGGTATTTTACATTAAAAATACCGCGCCTGCTGCTGAA
AAAAATCCATAAATGAAGTTTTGCCAGAGTCAGGACCAGACC
ATAAAAGGAATGGCCGACCGTGTGATAATGTTTAGAGGGCTT
CAATAAACATCCTAGTTGATATAAGTATAGAACCGCAAGCCC
AGGGAACTTTGTATTCCAACGTCAAAGGTCCGAAAGGCGAAA
TAAAGAATATAAATCCACGCTGGTTTGCAACGGTGAATCATA
ACGGCCACTTGGGCCGTATAACAACTAAGAGCCTAATGAGTG
AATCCCTCGTGGACCATCGCCTGATAAAGACGGTCTACAGAC
GCCCGAATAATACAATTGAGGAAGGTTATACTTCTAATATAA
CGCAAATATTCCATGCTGAATATAATGCGCCTCAGAGAATTA
GACTTTTTTCTCAAAAGAAGATGGTGGTGCGAAAAAAGTACA
ATCATTTTGAGTAGACTATCGGCCTTGCCTTTGAGTAAACGG
GAGGTTTGCCCTTTAATATCAGAGAGATTGAACACTTTTGTT
GTAATGTAGCTGATATAAAACGAACTAACCACATTAACACTA
TGCGTTAGAAATACTATTAGTCTTTAATTCACACGTTATTTA
AATTCATATCGCGTTAATTCACCGTCACAATAACGCCTTATT
AATCGGCTACCGCGAGCAAATCAGATATTTAACGGACAGTTA
CATTAAACAGCTGGGCCGGAAACCAGGCGATCGCATGGGATA
CAAAAGATCAAAAACTGGCCTTCCTGTATAACCGTGCATCAA
CTTAGAGTCGGTTGTAAATCATACAGGCGCATCAATACATTT
ATTCAGTCAAATCATTGATTATATTTTAGAGGGTATGGGAAG
CATTCAGCGACAGTAGATGGGCGCATCGGCCAGCTCAATAGG
TGCAGATTACGTTAAAATTAATGCCGGAAATGCAAATTTTTA
CAGAAGCAATGTCGAGTTTCTCCAACAGGGGGGTATACTGCG
GCGGGAGCCTCGTTGAGATTTAGGAATACGGAACATATTCAA
CGTAATCTAATAAAAACAATGCAACTAATACTCAGAACCGCC
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TACCTACAATCGCCTACTAGAAAAAGCCATAAGGCCAGACAA
GTAAAATCCCCAGCGTGAGGCCACCGAGTTAGTAAATCCAGA
TTTTAAGACGAGTATACCCAAATCAACGACAGGAATGTCAAT
CCCTAAAGAGAGCCGAAAAATCTAAAGCTATCAAACCGGCTT
GGAACGATACCGCCCAATACTTCTTTGATAAAAGACACTCAT
GCCTTGATATAGCACCCAAAAGTAATTCAGAAACCCGGATAA
CAGAACCCGAGAGGATTTACGAGCATGTTGTCCAGGTACCGA
ATCCTGTTCCTTTGACCAAATGAGGAAGCGAAACAACCGTCT
GAATCGTAGGTCTTATTGCATCAAAAAGAAGCGAATTAGAGA
GTGAATTATGGTTTTTGTCACAATCAATTAAGACTGAATACC
CCACGCTACATCGCGTTAATTTCATCTTTTAACCTATCATAG
TGATACAAAGAACGAAGCAAGCCGTTTTGAACGGGAAGAGAA
AGCTAACCGGTTTGGCCAGGGTGGTTTTGACGTTGTCCCCGG
CAAAAGGGTGCCACTCAGAAACAGTGCCTACCAGGAATCAAT
TATTTTTGGACATTCGCTCAATCGTCTGCAACAGTTATCATA
AGACGCTTATCAAAGTCAGTATTAACACAACGAACAACTTTT
CCGTTCTGTAGGTATATTTCAACGCAAGTAAGAGCCAACTAA
GAATATAACAATTTCTTACCAACGCTAATACAGAGAGAATCA
GGTCACGATGTGAGTAAATCAGCTCATTCGGGCCTCATTCGC
GAGTTAAGCATTAGAAAATAGCAGCCTTCGAGCGTTTTTGCA
TAACGTCATTTGCCTTAAATCAAGATTAAGCAATAAAGAATT
AACAGTTTTAGAACATCAGATGATGGCACCACCAGATCCTTT
GGGAAGCGCCCAATAGAAACAATGAAATGTTGCTACTTTCCA
ACGCAGTGTTTATTACCAGCGCCAAAGAATTATTCAGCCATT
TCAAATAGTTATATGTCAATAGTGAATTATCACCTAACAGTG
ATGCCCCGATTAGCCCTTATCATTCCAATATTTTCTTATCCG
ACTTGCCATAATCAGATTATACCAAGCGTTTCCATGACTAAA
GGAATAAATGTTAGCGAGGAAACGCAATCGACTTGATTAAAG
GAACCCTGAAAGGAGCCAAATAAGGCTTAGGACGTGCTATTT
GCAAAATGGGGCGCAACCTGTTTAGCTAAGTACGGCAACATG
GTACCGACGAGCCGAATTGCGTTGCGCTCGGCCAAACCAGTG
AACATCAGCTGAATTATCCTTCTGTAAAGTTTAACCATCGGG
GAGCCTTCGCATAACGGTCGCTGAGGCTAGCCCTCGAATTTT
CATATGTATCTACATCATTATACCAGTCGCCCTGAGCTGCTC
TCGCCCATAATTGTTTTCACGTTGAAAAAGTAAATATAGTTA
CAACATAGCTCGAAGTTTTCCCAGTCACTCTTTTCCGCGCGG
CAATAGACGTTATTGCGGAACAAAGAAAATTCATCGAATAAT
ACGGAGACGAACTGAAAGAGGACAGATGCCCAGCATCGGCAA
TCCTGATTCATTTTAATTTTAAAAGTTTTAGATTATATCTTT
CATTGGCAGCCAGTTCTTAACTACCTTTCTGACCTCTGATAG
TTGAGAGACCCCGGGAAAAGCCCCAAAATAACAAACGAGAAA
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ACTACAATTTGCGGATAGTTGCGCCGACTTGCTTTAGGAATT
GAATCATTGTCTTTGGGGTTTTGCTCAGCGTATAAGGTCAGT
TAACCTTAGAAAACAAAGAAGATGATGACCTTTTAGTCAGAT
AACCGGAATATTTAACGGTAATCGTAAAATTGCCTTATGCGA
ATGGTTTTACAAATATAAAGCCAACGCTAAATGGAACCAGTA
CCCAGCTCAGAGCAAATAAGTAACAGTAAAAACATAAAAACA
TACCAGATAGAGCCAAATATTGACGGAACAAAAGGACACCAC
CAATTACATAACGGGCGTAGATTTTCAGTCGTCGCGAGTGAA
ACCCTCAATTCACATCCTCATTAAAGCCATCGATAGTAGCGC
GTTTTCAACCACCGCACCCTCAGAGCCGGGTCAGAAAAGCGC
CTGTATGATAATTTATCGGTTTATCAGCAATGACAAGTTAAA
GAAATAACCCTTAGAGTACATAAATCAATTAACAATTCATTT
AACATTATGCATGCGCTTGTTTGGATTATCTAAAAGAGCCGT
AATGTTTGATCTAATCTTTCCAGACGTTTCTCCAAAAAAAGT
CTTTGACACGTAATAACGGCTACAGAGGTGGTAATTAACATC
AGGCTTTTAATTGCTAATTCGAGCTTCAATTAAGAATAAATC
AGGAGCAGTGCCAACTGCAGGTCGACTCAAAGCCTGGGGTGT
GCGAATAGGATTTTAGCATTCCACAGACTGCAGGGACAACCA
GCGTAACAGATATTCCGATACTGGATAGTAAGGCTCCAAAAG
TTGCAGCATAGGCTCGGAACGAGGCGCATTGTGTCCCACTAT
GAGGCATACCCATGGTACCGCCACCCTCAGCCCGGGAAAAAT
TTTTAAAAGTAGCGGTTTGCCTTTAGCGAGAGCCGCCACCTT
CAGAGCCTCGGCATTCACCAATGAAACCAGAATGGCGATTGG
CCTTGATGAGCCACGAACCGCCTCCCTCTCAGACTGCAGCAC
AAGTTTCGGTCAATGAGCTGAAAAGGTGAAGGCAAAGCATAA
GGATTAGCTGCCTAACTGGTAATAAGTTAGAAGGCATCGTAG
GGGACGAGCTGCGCAAGGGCGATCGGTGTTTTAACTTCATCA
AACGCCAACTTGGGAACTGTGGCATGATAGTGCCAGTTTGAG
CACCAGAAACTGGCAAGGCTATCAGGTCACTAGCAGATTGTA
GTCTGAGAGCAAATAGCAGATTCACCAGGCGCGAAAAATTTA
GGAAGCAAAAGCGGTACCCTGACTATTACGTCCAAATAGTAA
TTCATTTTAATCAAACAGAAGCAATAAATGTAGCTTGTCTGG
GTGCCGTGCCACCCCACCCTCATTTTCAATATAAAACGACGA
ACATTAATTGGTGTATCGGCCTCAGGAAAAAGCGCCTTCGCT
CAAGGCCGAATTTAAGGAGGTTGAGGCACCACCAGCCGGAAC
AATATCCCAACATGCCAGTAATAAGAGAGGGATAGCCACCCT
AGAAACACTGAGCAAAAATTAATTACATTATATGTTATTAAT
TGGGTAACAACAGCCTGCATTAATGAATCACTGCCTTCCACA
CCTCAAAGAGAAGAAACTATATGTAAATGCGAGAACACCAGCAGAAGATAAAAC
TCATAACAAGCCTTAAGATTCAAAAGGGAATATGAACATTATTACAGGTAGAAA
AGCTAAACTTAATTATAACAGTTGATTCCATTAGATTCTACTAATAGTAGTAGC
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AATAGGATTTCGAGTTCAGCTAATGCAGTGAACAAAATAGGTGTATCACCGTAC
GGAAGGGGAAAGGATTTGAGGATTTAGAGTATTAAAAGGAGCGGAATTATCATC
TACCGAATGAAGCCAGTTACAAAATAAAAACGATTCCTGAACAAAGTCAGAGGG
GTACCTTTGCAAAAATTCATTGAATCCCAATGACCGGAAGCCCGAAAGACTTCA
TAAGCAATATTCATGTAAATTGGGCTTGATTACCTGAGAGTCTGGAGCAAACAA
CAGGCGCAAGCGGTCAAAAGAATAGCCCCAAGAGTGAAATCCGCGACCTGCTCC
AGACGGGCGCCAGGTTCGTAATCATGGTCTCACAACGCTTTCCAGTCGGGAAAC
GGCCGCTCGCCTGTGCTAAACAACTTTCCAACTAACGAGGTGAATTTCTTAAAC
AGTCTCTGGAAACGTTTCGGTCATAGCCAAAATCAAACCACCACCAGAGCCGCC
AATTGAGATTTTGACTGGCCAACAGAGACGTGGCAGTTAAATAAGAATAAACAC
GTATTCTGGAGTGTTTTCGGAACCTATTACTCCTCTATTAAACCAAGTACCGCA
TGGGAATAGGAAACCAAACGTAGAAAATCGCAAAGGCGACATTCAACCGATTGA
ACAATATGGGTAGCGCCACTACGAAGGCACTAAAAGTCTGTCCATCACGCAAAT
ATTACGCTTTTTGTCGAGTAACAACCCGACCGTAACTCCAGCCAGCTTTCCGGC
TAATTTTAGAAATTATTCGCCTGATTGCCGAATTATTTCATTTGAATTACCTTT
TAACCGTTGTAGAGCCATTGCAACAGCGAAAGATAAAACGAATTATGAAGGT
TTTAATGGAAACAATCCTTGAAAACATTGCACGACCAAGTTATTATGAAGGT
CTGTCGTGCCAGTGATTGCCCTTCACCAAGGCGGTTTCCTGTTTATGAAGGT
AGCTTGATACCGGATCGTCACCCTCACGTCACCTCAGCGGAGTTATGAAGGT
GATTCATCAGTTTACCAGACGACGATCCTGTAAGGCCGGAGATTATGAAGGT
CGGAATCATAATATATTTAACAACGCAACGCTCCTTCTGACCTTATGAAGGT
GCCAGCATTGACCCGTTCCAGTAAGCACCATTATAGCGTTTGTTATGAAGGT
ATGTTACTTAGCGGCTGACCTTCATCTGGCCCTGGTTGAGTGTTATGAAGGT
TAATTGAGCGCTTTAAGAAAAGTAAGCTCCCGAATTATTTATTTATGAAGGT
TCAGGAGGTTTATACCGTAACACTGAATGTAATCTGTTTATCTTATGAAGGT
CTCATCGAGAACCGAGGCGTTTTAGCTACATGGAACATGAAATTATGAAGGT
GAGAATCGATGAAATTGTAAACGTTAGTAATCTTTAATTTCATTATGAAGGT
ATATTCCTGATTCTACCATATCAAAATCAGTTGGACTTTACATTATGAAGGT
ATTAACATCCAATACCAAAAACATTAATTTTTGTGCGAACGATTATGAAGGT
AATATCGCGTTTTCCTTTTGATAAGAACCAAAATGCTTTAAATTATGAAGGT
ACCGCTTCTGGTCGAAAGGGGGATGTTTTGTTACTCCGTGGGTTATGAAGGT
GGGAGGGAAGGTAGCAAAATCACCAGTAGCCGATAAAGGTGGTTATGAAGGT
AGAGGTGAGGCGCCCTCAATCAATATGATAGCTCAAATCCAATTATGAAGGT
© 2016 Macmillan Publishers Limited. All rights reserved.
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Reference
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© 2016 Macmillan Publishers Limited. All rights reserved.