| 质粒名称: | pSimpleII-U6-tracr-U6-BsmBI-NLS-NmCas9-HA-NLS(s) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 47868 |
| 存储实验室: | Erik Sontheimer, James Thomson |
|
载体骨架基本信息 |
|
| 载体名称: | pSimpleII |
| 载体抗性: | 氨苄西林 |
| 空载体大小: | 5000bp |
| 完整质粒大小: | 8051 bp |
| 载体类型: | 哺乳细胞表达 |
| 生长条件: | 37 ℃ |
|
插入基因1信息 |
|
| 插入基因名称: | NmCas9 |
| 备用名称： | CSN1 |
| 物种来源: | 脑膜炎N meningitidis |
| 插入大小: | 3243 bp |
| 启动子 | EF1a |
| 标签/融合蛋白: |
NLS（插入件上的N端子）
NLS（插入件上的C端子）
HA（插入件上的C端子）
|
|
插入基因2信息 |
|
| 插入基因名称： | U6pr-tracrRNA |
| 物种来源： | 脑膜炎N meningitidis |
| 插入大小： | 364 |
| 启动子： | U6 |
|
文献引用方法 |
|
| 材料和方法部分: | pSimpleII-U6-tracr-U6-BsmBI-NLS-NmCas9-HA-NLS(s) was a gift from Erik Sontheimer & James Thomson (Addgene plasmid # 47868 ; http://n2t.net/addgene:47868 ; RRID:Addgene_47868) |
| 参考文献部分: | Efficient genome engineering in human pluripotent stem cells using Cas9 from Neisseria meningitidis. Hou Z, Zhang Y, Propson NE, Howden SE, Chu LF, Sontheimer EJ, Thomson JA. Proc Natl Acad Sci U S A. 2013 Aug 12. 10.1073/pnas.1313587110 PubMed 23940360 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
质粒图谱.png)
载体简介
载体序列
LOCUS pSimpleII-U6-tracr-U6-BsmBI-NLS-NmCas9-HA-NLS(s) 9258 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION This plasmid contains expression cassette for NmCas9 with N and C
NLS and an HA tag, a cassette for expression of tracrRNA, a cassette
for cloning crRNA under the control of U6 promoter..
ACCESSION .
VERSION .
KEYWORDS pSimpleII-U6-tracr-U6-BsmBI-NLS-NmCas9-HA-NLS(s)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9258)
AUTHORS Hou Z, Zhang Y, Propson NE, Howden SE, Chu LF, Sontheimer EJ,
Thomson JA
TITLE Efficient genome engineering in human pluripotent stem cells using
Cas9 from Neisseria meningitidis.
JOURNAL Proc Natl Acad Sci U S A. 2013 Aug 12.
PUBMED 23940360
REFERENCE 2 (bases 1 to 9258)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9258
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter complement(44..284)
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 322..340
/label=T7 Term
/note="T7 terminator, reverse primer"
promoter complement(359..377)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_RNA complement(419..511)
/label=Nm tracrRNA
/note="trans-activating CRISPR RNA for the Neisseria
meningitidis CRISPR/Cas9 system (Hou et al., 2013)"
promoter complement(519..759)
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind complement(824..843)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 993..2166
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 1224..2157
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 2114..2134
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
regulatory 2172..2181
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2184..2204
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 2205..5447
/codon_start=1
/product="Cas9 endonuclease from the Neisseria meningitidis
Type II CRISPR/Cas system (Esvelt et al., 2013)"
/label=NmCas9
/note="generates RNA-guided double strand breaks in DNA"
/translation="AAFKPNSINYILGLDIGIASVGWAMVEIDEEENPIRLIDLGVRVF
ERAEVPKTGDSLAMARRLARSVRRLTRRRAHRLLRTRRLLKREGVLQAANFDENGLIKS
LPNTPWQLRAAALDRKLTPLEWSAVLLHLIKHRGYLSQRKNEGETADKELGALLKGVAG
NAHALQTGDFRTPAELALNKFEKESGHIRNQRSDYSHTFSRKDLQAELILLFEKQKEFG
NPHVSGGLKEGIETLLMTQRPALSGDAVQKMLGHCTFEPAEPKAAKNTYTAERFIWLTK
LNNLRILEQGSERPLTDTERATLMDEPYRKSKLTYAQARKLLGLEDTAFFKGLRYGKDN
AEASTLMEMKAYHAISRALEKEGLKDKKSPLNLSPELQDEIGTAFSLFKTDEDITGRLK
DRIQPEILEALLKHISFDKFVQISLKALRRIVPLMEQGKRYDEACAEIYGDHYGKKNTE
EKIYLPPIPADEIRNPVVLRALSQARKVINGVVRRYGSPARIHIETAREVGKSFKDRKE
IEKRQEENRKDREKAAAKFREYFPNFVGEPKSKDILKLRLYEQQHGKCLYSGKEINLGR
LNEKGYVEIDHALPFSRTWDDSFNNKVLVLGSENQNKGNQTPYEYFNGKDNSREWQEFK
ARVETSRFPRSKKQRILLQKFDEDGFKERNLNDTRYVNRFLCQFVADRMRLTGKGKKRV
FASNGQITNLLRGFWGLRKVRAENDRHHALDAVVVACSTVAMQQKITRFVRYKEMNAFD
GKTIDKETGEVLHQKTHFPQPWEFFAQEVMIRVFGKPDGKPEFEEADTLEKLRTLLAEK
LSSRPEAVHEYVTPLFVSRAPNRKMSGQGHMETVKSAKRLDEGVSVLRVPLTQLKLKDL
EKMVNREREPKLYEALKARLEAHKDDPAKAFAEPFYKYDKAGNRTQQVKAVRVEQVQKT
GVWVRNHNGIADNATMVRVDVFEKGDKYYLVPIYSWQVAKGILPDRAVVQGKDEEDWQL
IDDSFNFKFSLHPNDLVEVITKKARMFGYFASCHRGTGNINIRIHDLDHKIGKNGILEG
IGVKTALSFQKYQIDELGKEIRPCRLKKRPPVR"
CDS 5448..5474
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 5481..5507
/codon_start=1
/product="nuclear localization signal (Makkerh et al.,
1996)"
/label=NLS
/translation="PAAKKKKLD"
primer_bind complement(5523..5540)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5529..5753
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 5942..7055
/label=mini-oriP
/note="internally truncated variant of the Epstein-Barr
virus oriP replication origin (Tanaka et al., 1999)"
primer_bind complement(7101..7117)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7101..7117)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(7114..7136)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 7125..7141
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(7149..7179)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7194..7215
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(7332..7349)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(7503..8091)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7583..7602)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(8262..9122)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 8885..8904
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(9123..9227)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gtttaaacaa aaaaatgaga cgtggaatcg tctccggtgt ttcgtccttt ccacaagata
61 tataaagcca agaaatcgaa atactttcaa gttacggtaa gcatatgata gtccatttta
121 aaacataatt ttaaaactgc aaactaccca agaaattatt actttctacg tcacgtattt
181 tgtactaata tctttgtgtt tacagtcaaa ttaattctaa ttatctctct aacagccttg
241 tatcgtatat gcaaatatga aggaatcatg ggaaataggc cctcttcctg cccgacctta
301 gatccgaatc catatcgata tgctagttat tgctcagcgg tggcagcagc caactcagcc
361 tatagtgagt cgtattaatt tcgcgggatg tttaaacgcg gccgcaaaaa aaccgaaata
421 aacgatgccc cttaaagcag aagctttaag gggcagagcg ttgcggcaca tcttttcaga
481 cggccttatt gtagcaacgg ttctcatttc ggtgtttcgt cctttccaca agatatataa
541 agccaagaaa tcgaaatact ttcaagttac ggtaagcata tgatagtcca ttttaaaaca
601 taattttaaa actgcaaact acccaagaaa ttattacttt ctacgtcacg tattttgtac
661 taatatcttt gtgtttacag tcaaattaat tctaattatc tctctaacag ccttgtatcg
721 tatatgcaaa tatgaaggaa tcatgggaaa taggccctct tcctgcccga ccttgacgtc
781 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
841 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
901 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
961 ttagggttag gcgttttgcg ctgcttcgcg gaggctccgg tgcccgtcag tgggcagagc
1021 gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct
1081 agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc
1141 ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca
1201 acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct
1261 ttacgggtta tggcccttgc gtgccttgaa ttacttccac ctggctccag tacgtgattc
1321 ttgatcccga gctggagcca ggggcgggcc ttgcgcttta ggagcccctt cgcctcgtgc
1381 ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg
1441 cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc
1501 gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caggatctgc acactggtat
1561 ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc
1621 gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtcggacg ggggtagtct
1681 caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg
1741 gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc
1801 cctgctccag ggggctcaaa atggaggacg cggcgctcgg gagagcgggc gggtgagtca
1861 cccacacaaa ggaaaggggc ctttccgtcc tcagccgtcg cttcatgtga ctccacggag
1921 taccgggcgc cgtccaggca cctcgattag ttctggagct tttggagtac gtcgtcttta
1981 ggttgggggg aggggtttta tgcgatggag tttccccaca ctgagtgggt ggagactgaa
2041 gttaggccag cttggcactt gatgtaattc tccttggaat ttgccctttt tgagtttgga
2101 tcttggttca ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc atttcaggtg
2161 tcgtgaacac cgccaccatg gtgcctaaga agaagagaaa ggtggctgcc ttcaaaccta
2221 attcaatcaa ctacatcctc ggcctcgata tcggcatcgc atccgtcggc tgggcgatgg
2281 tagaaattga cgaagaagaa aaccccatcc gcctgattga tttgggcgtg cgcgtatttg
2341 agcgtgccga agtaccgaaa acaggcgact cccttgccat ggcaaggcgt ttggcgcgca
2401 gtgttcgccg cctgacccgc cgtcgcgccc accgcctgct tcggacccgc cgcctattga
2461 aacgcgaagg cgtattacaa gccgccaatt ttgacgaaaa cggcttgatt aaatccttac
2521 cgaatacacc atggcaactt cgcgcagccg cattagaccg caaactgacg cctttagagt
2581 ggtcggcagt cttgttgcat ttaatcaaac atcgcggcta tttatcgcaa cggaaaaacg
2641 agggcgaaac tgccgataag gagcttggcg ctttgcttaa aggcgtagcc ggcaatgccc
2701 atgccttaca gacaggcgat ttccgcacac cggccgaatt ggctttaaat aaatttgaga
2761 aagaaagcgg ccatatccgc aatcagcgca gcgattattc gcatacgttc agccgcaaag
2821 atttacaggc ggagctgatt ttgctgtttg aaaaacaaaa agaatttggc aatccgcatg
2881 tttcaggcgg ccttaaagaa ggtattgaaa ccctactgat gacgcaacgc cctgccctgt
2941 ccggcgatgc cgttcaaaaa atgttggggc attgcacctt cgaaccggca gagccgaaag
3001 ccgctaaaaa cacctacaca gccgaacgtt tcatctggct gaccaagctg aacaacctgc
3061 gtattttaga gcaaggcagc gagcggccat tgaccgatac cgaacgcgcc acgcttatgg
3121 acgagccata cagaaaatcc aaactgactt acgcacaagc ccgtaagctg ctgggtttag
3181 aagataccgc ctttttcaaa ggcttgcgct atggtaaaga caatgccgaa gcctcaacat
3241 tgatggaaat gaaggcctac catgccatca gccgtgcact ggaaaaagaa ggattgaaag
3301 acaaaaaatc cccattaaac ctttctcccg aattacaaga cgaaatcggc acggcattct
3361 ccctgttcaa aaccgatgaa gacattacag gccgtctgaa agaccgtata cagcccgaaa
3421 tcttagaagc gctgttgaaa cacatcagct tcgataagtt cgtccaaatt tccttgaaag
3481 cattgcgccg aattgtgcct ctaatggaac aaggcaaacg ttacgatgaa gcctgcgccg
3541 aaatctacgg agaccattac ggcaagaaga atacggaaga aaagatttat ctgccgccga
3601 ttcccgccga cgaaatccgc aaccccgtcg tcttgcgcgc cttatctcaa gcacgtaagg
3661 tcattaacgg cgtggtacgc cgttacggct ccccagctcg tatccatatt gaaactgcaa
3721 gggaagtagg taaatcgttt aaagaccgca aagaaattga gaaacgccaa gaagaaaacc
3781 gcaaagaccg ggaaaaagcc gccgccaaat tccgagagta tttccccaat tttgtcggag
3841 aacccaaatc caaagatatt ctgaaactgc gcctgtacga gcaacaacac ggcaaatgcc
3901 tgtattcggg caaagaaatc aacttaggcc gtctgaacga aaaaggctat gtcgaaatcg
3961 accatgccct gccgttctcg cgcacatggg acgacagttt caacaataaa gtactggtat
4021 tgggcagcga aaaccaaaac aaaggcaatc aaacccctta cgaatacttc aacggcaaag
4081 acaacagccg cgaatggcag gaatttaaag cgcgtgtcga aaccagccgt ttcccgcgca
4141 gtaaaaaaca acggattctg ctgcaaaaat tcgatgaaga cggctttaaa gaacgcaatc
4201 tgaacgacac gcgctacgtc aaccgtttcc tgtgtcaatt tgttgccgac cgtatgcggc
4261 tgacaggtaa aggcaagaaa cgtgtctttg catccaacgg acaaattacc aatctgttgc
4321 gcggcttttg gggattgcgc aaagtgcgtg cggaaaacga ccgccatcac gccttggacg
4381 ccgtcgtcgt tgcctgctcg accgttgcca tgcagcagaa aattacccgt tttgtacgct
4441 ataaagagat gaacgcgttt gacggtaaaa ccatagacaa agaaacagga gaagtgctgc
4501 atcaaaaaac acacttccca caaccttggg aatttttcgc acaagaagtc atgattcgcg
4561 tcttcggcaa accggacggc aaacccgaat tcgaagaagc cgatacccta gaaaaactgc
4621 gcacgttgct tgccgaaaaa ttatcatctc gccccgaagc cgtacacgaa tacgttacgc
4681 cactgtttgt ttcacgcgcg cccaatcgga agatgagcgg gcaagggcat atggagaccg
4741 tcaaatccgc caaacgactg gacgaaggcg tcagcgtgtt gcgcgtaccg ctgacacagt
4801 taaaactgaa agacttggaa aaaatggtca atcgggagcg cgaacctaag ctatacgaag
4861 cactgaaagc acggctggaa gcacataaag acgatcctgc caaagccttt gccgagccgt
4921 tttacaaata cgataaagca ggcaaccgca cccaacaggt aaaagccgta cgcgtagagc
4981 aagtacagaa aaccggcgta tgggtgcgca accataacgg tattgccgac aacgcaacca
5041 tggtgcgcgt agatgtgttt gagaaaggcg acaagtatta tctggtaccg atttacagtt
5101 ggcaggtagc gaaagggatt ttgccggata gggctgttgt acaaggaaaa gatgaagaag
5161 attggcaact tattgatgat agtttcaact ttaaattctc attacaccct aatgatttag
5221 tcgaggttat aacaaaaaaa gctagaatgt ttggttactt tgccagctgc catcgaggca
5281 caggtaatat caatatacgc attcatgatc ttgatcataa aattggcaaa aatggaatac
5341 tggaaggtat cggcgtcaaa accgcccttt cattccaaaa ataccaaatt gacgaactgg
5401 gcaaagaaat cagaccatgc cgtctgaaaa aacgcccgcc tgtccgttac ccatacgatg
5461 ttccagatta cgctgcagct ccagcagcga agaaaaagaa gctggattaa ctcgctgatc
5521 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
5581 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
5641 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
5701 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
5761 ggcggaaaga accagctggg gctcgaccga tgcccttgag agccttcaac ccagtcagct
5821 ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca
5881 tgcaactcgt aggacaggtg cctggccggg gtcccccgga aactcggccg tggtgaccat
5941 gcaggaaaag gacaagcagc gaaaattcac gcccccttgg gaggtggcgg catatgcaaa
6001 ggatagcact cccactctac tactgggtat catatgctga ctgtatatgc atgaggatag
6061 catatgctac ccggatacag attaggatag catatactac ccagatatag attaggatag
6121 catatgctac ccagatatag attaggatag cctatgctac ccagatataa attaggatag
6181 catatactac ccagatatag attaggatag catatgctac ccagatatag attaggatag
6241 cctatgctac ccagatatag attaggatag catatgctac ccagatatag attaggatag
6301 catatgctat ccagatattt gggtagtata tgctacccag atataaatta ggatagcata
6361 tactacccta atctctatta ggatagcata tgctacccgg atacagatta ggatagcata
6421 tactacccag atatagatta ggatagcata tgctacccag atatagatta ggatagccta
6481 tgctacccag atataaatta ggatagcata tactacccag atatagatta ggatagcata
6541 tgctacccag atatagatta ggatagccta tgctacccag atatagatta ggatagcata
6601 tgctatccag atatttgggt agtatatgct acccatggca acattagccc accgtgctct
6661 cagcgacctc gtgaatatga ggaccaacaa ccctgtgctt ggcgctcagg cgcaagtgtg
6721 tgtaatttgt cctccagatc gcagcaatcg cgcccctatc ttggcccgcc cacctactta
6781 tgcaggtatt ccccggggtg ccattagtgg ttttgtgggc aagtggtttg accgcagtgg
6841 ttagcggggt tacaatcagc caagttatta cacccttatt ttacagtcca aaaccgcagg
6901 gcggcgtgtg ggggctgacg cgatcgctgt tccttaggac ccttttacta accctaattc
6961 gatagcatat gcttcccgtt gggtaacata tgctattgaa ttagggttag tctggatagt
7021 atatactact acccgggaag catatgctac ccgtttaggg ttataccgtc gacctctagc
7081 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
7141 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
7201 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
7261 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
7321 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
7381 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
7441 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
7501 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
7561 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
7621 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
7681 agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
7741 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
7801 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
7861 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
7921 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
7981 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
8041 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
8101 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
8161 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
8221 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
8281 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
8341 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
8401 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
8461 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
8521 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
8581 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
8641 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
8701 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
8761 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
8821 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
8881 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
8941 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
9001 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
9061 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
9121 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
9181 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
9241 aaagtgccac ctgacgtc
//
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