| 质粒名称: | pSpCas9n(BB) 、pX460 |
|---|---|
| 载体又名: | pSpCas9n(BB) 载体 、pX460载体 、pSpCas9n(BB) 质粒 、pX460 质粒 |
| 出品公司: | Addgene |
| 目录编号: | 48873 |
| 存储实验室: | Feng Zhang |
|
载体骨架基本信息 |
|
| 载体名称: | PX460 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄西林 |
| 空载体大小: | 8500 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达,CRISPR |
| 生长菌株: | Stbl3 |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | hSpCas9n |
| 物种来源: | 人工合成 |
| 插入大小: | 5400 bp |
| 基因突变: | Cas9 D10A nickase mutant |
| GenBank编号 | -- |
| 启动子 | Cbh |
| 标签/融合蛋白: | 3XFLAG（插入N端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | EcoRI(未破坏) |
| 5' 测序引物 | TTTATGGCGAGGCGGCGG |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pSpCas9n(BB) (PX460) was a gift from Feng Zhang (Addgene plasmid # 48873 ; http://n2t.net/addgene:48873 ; RRID:Addgene_48873) |
| 参考文献部分: | Genome engineering using the CRISPR-Cas9 system. Ran FA, Hsu PD, Wright J, Agarwala V, Scott DA, Zhang F. Nat Protoc. 2013 Nov;8(11):2281-308. doi: 10.1038/nprot.2013.143. Epub 2013 Oct 24. 10.1038/nprot.2013.143 PubMed 24157548 |
买家导航
pSpCas9n(BB)载体基本信息
pSpCas9n(BB)载体图谱质粒图谱和多克隆位点信息
 (PX460)质粒图谱.png)
pSpCas9n(BB)载体简介
pSpCas9n(BB)载体序列
LOCUS pSpCas9n(BB) (PX460) 8505 bp ds-DNA circular SYN 21-AUG-2019
DEFINITION Cas9n (D10A nickase mutant) from S. pyogenes.
ACCESSION .
VERSION .
KEYWORDS pSpCas9n(BB) (PX460)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8505)
AUTHORS Ran FA, Hsu PD, Wright J, Agarwala V, Scott DA, Zhang F
TITLE Genome engineering using the CRISPR-Cas9 system.
JOURNAL Nat Protoc. 2013 Nov;8(11):2281-308. doi: 10.1038/nprot.2013.143.
Epub 2013 Oct 24.
PUBMED 24157548
REFERENCE 2 (bases 1 to 8505)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 21, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8505
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..241
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 1..21
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 172..191
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 268..343
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
enhancer 440..725
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer;
contains an 18-bp deletion relative to the standard CMV
enhancer"
promoter 727..1004
/label=chicken beta-actin promoter
intron 1005..1232
/label=hybrid intron
/note="hybrid between chicken beta-actin (CBA) and minute
virus of mice (MMV) introns (Gray et al., 2011)"
regulatory 1244..1253
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1253..1318
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 1325..1345
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1370..5470
/codon_start=1
/product="nickase mutant of the Cas9 endonuclease from the
Streptococcus pyogenes Type II CRISPR/Cas system"
/label=Cas9(D10A)
/note="generates RNA-guided single strand nicks in DNA due
to the D10A mutation in the RuvC catalytic domain"
/translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 5471..5518
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
primer_bind complement(5546..5563)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5552..5759
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 5768..5908
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 5768..5897
/label=AAV2 ITR
rep_origin 5983..6438
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6070..6089)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6280..6301
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(6455..6474)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6574..6596
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(6634..6652)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6720..6824
/gene="bla"
/label=AmpR promoter
CDS 6825..7685
/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 complement(7043..7062)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7856..8444
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8345..8364
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
241 cgaaacaccg ggtcttcgag aagacctgtt ttagagctag aaatagcaag ttaaaataag
301 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg ttttagagct
361 agaaatagca agttaaaata aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac
421 aaatggctct agaggtaccc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
481 ccaacgaccc ccgcccattg acgtcaatag taacgccaat agggactttc cattgacgtc
541 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
601 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tgtgcccagt
661 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
721 ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac
781 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg
841 ggggggggcg cgcgccaggc ggggcggggc ggggcgrggg gcggggcggg gcgaggcgga
901 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc
961 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct
1021 gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga
1081 ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc
1141 tgagcaagag gtaagggttt aagggatggt tggttggtgg ggtattaatg tttaattacc
1201 tggagcacct gcctgaaatc actttttttc aggttggacc ggtgccacca tggactataa
1261 ggaccacgac ggagactaca aggatcatga tattgattac aaagacgatg acgataagat
1321 ggccccaaag aagaagcgga aggtcggtat ccacggagtc ccagcagccg acaagaagta
1381 cagcatcggc ctggccatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta
1441 caaggtgccc agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa
1501 gaacctgatc ggagccctgc tgttcgacag cggcgaaaca gccgaggcca cccggctgaa
1561 gagaaccgcc agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat
1621 cttcagcaac gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt
1681 cctggtggaa gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga
1741 ggtggcctac cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag
1801 caccgacaag gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg
1861 gggccacttc ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt
1921 catccagctg gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg
1981 cgtggacgcc aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct
2041 gatcgcccag ctgcccggcg agaagaagaa tggcctgttc ggcaacctga ttgccctgag
2101 cctgggcctg acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca
2161 gctgagcaag gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca
2221 gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat
2281 cctgagagtg aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata
2341 cgacgagcac caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga
2401 gaagtacaaa gagattttct tcgaccagag caagaacggc tacgccggct acattgacgg
2461 cggagccagc caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg
2521 caccgaggaa ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt
2581 cgacaacggc agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg
2641 gcaggaagat ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac
2701 cttccgcatc ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat
2761 gaccagaaag agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg
2821 cgcttccgcc cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga
2881 gaaggtgctg cccaagcaca gcctgctgta cgagtacttc accgtgtata acgagctgac
2941 caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa
3001 aaaggccatc gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa
3061 agaggactac ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga
3121 tcggttcaac gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga
3181 cttcctggac aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact
3241 gtttgaggac agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga
3301 caaagtgatg aagcagctga agcggcggag atacaccggc tggggcaggc tgagccggaa
3361 gctgatcaac ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc
3421 cgacggcttc gccaacagaa acttcatgca gctgatccac gacgacagcc tgacctttaa
3481 agaggacatc cagaaagccc aggtgtccgg ccagggcgat agcctgcacg agcacattgc
3541 caatctggcc ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga
3601 cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg aaatggccag
3661 agagaaccag accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga
3721 agagggcatc aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca
3781 gctgcagaac gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga
3841 ccaggaactg gacatcaacc ggctgtccga ctacgatgtg gaccatatcg tgcctcagag
3901 ctttctgaag gacgactcca tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg
3961 caagagcgac aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcggca
4021 gctgctgaac gccaagctga ttacccagag aaagttcgac aatctgacca aggccgagag
4081 aggcggcctg agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg
4141 gcagatcaca aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga
4201 gaatgacaag ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga
4261 tttccggaag gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca
4321 cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga
4381 aagcgagttc gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag
4441 cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc tacagcaaca tcatgaactt
4501 tttcaagacc gagattaccc tggccaacgg cgagatccgg aagcggcctc tgatcgagac
4561 aaacggcgaa accggggaga tcgtgtggga taagggccgg gattttgcca ccgtgcggaa
4621 agtgctgagc atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt
4681 cagcaaagag tctatcctgc ccaagaggaa cagcgataag ctgatcgcca gaaagaagga
4741 ctgggaccct aagaagtacg gcggcttcga cagccccacc gtggcctatt ctgtgctggt
4801 ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag agctgctggg
4861 gatcaccatc atggaaagaa gcagcttcga gaagaatccc atcgactttc tggaagccaa
4921 gggctacaaa gaagtgaaaa aggacctgat catcaagctg cctaagtact ccctgttcga
4981 gctggaaaac ggccggaaga gaatgctggc ctctgccggc gaactgcaga agggaaacga
5041 actggccctg ccctccaaat atgtgaactt cctgtacctg gccagccact atgagaagct
5101 gaagggctcc cccgaggata atgagcagaa acagctgttt gtggaacagc acaagcacta
5161 cctggacgag atcatcgagc agatcagcga gttctccaag agagtgatcc tggccgacgc
5221 taatctggac aaagtgctgt ccgcctacaa caagcaccgg gataagccca tcagagagca
5281 ggccgagaat atcatccacc tgtttaccct gaccaatctg ggagcccctg ccgccttcaa
5341 gtactttgac accaccatcg accggaagag gtacaccagc accaaagagg tgctggacgc
5401 caccctgatc caccagagca tcaccggcct gtacgagaca cggatcgacc tgtctcagct
5461 gggaggcgac aaaaggccgg cggccacgaa aaaggccggc caggcaaaaa agaaaaagta
5521 agaattccta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
5581 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
5641 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
5701 ggggtggggc aggacagcaa gggggaggat tgggaagaga atagcaggca tgctggggag
5761 cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
5821 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
5881 agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat
5941 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
6001 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct
6061 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
6121 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
6181 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
6241 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
6301 gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
6361 cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
6421 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
6481 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
6541 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
6601 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
6661 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
6721 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
6781 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
6841 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
6901 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
6961 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
7021 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
7081 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
7141 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
7201 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
7261 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
7321 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
7381 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
7441 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
7501 gctggtttat tgctgataaa tctggagccg gtgagcgtgg aagccgcggt atcattgcag
7561 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
7621 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
7681 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
7741 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
7801 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
7861 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
7921 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
7981 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
8041 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
8101 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
8161 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
8221 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
8281 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
8341 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
8401 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
8461 cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
