| 质粒名称: | pX330-U6-Chimeric_BB-CBh-hSpCas9-hGem(1/110) |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 71707 |
| 存储实验室: | Giulio Draetta |
|
载体骨架基本信息 |
|
| 载体名称: | pX330-U6-Chimeric_BB-CBh-hSpCas9 |
| 载体出品公司: | Zhang lab, Addgene plasmid 42230 |
| 载体抗性: | 氨苄青霉素 |
| 完整质粒大小: | 8854 bp |
| 载体类型: | 哺乳细胞表达,CRISPR |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | humanized S. pyogenes Cas9 fused to human Geminin,hSpCas9-hGem(1/110),hCas9-hGem(1/110),GEMININ |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 4614 bp |
| 启动子 | CBh |
| 标签/融合蛋白: |
3xFLAG（插入件上的N端子）
hGEM（1/110）（插入件上的C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 | pCBhProF (AGGGATGGTTGGTTGGTGGG) |
| 3' 测序引物 | CCAATCCTCCCCCTTGCTGT or BGH-rev |
|
文献引用方法 |
|
| 材料和方法部分: | pX330-U6-Chimeric_BB-CBh-hSpCas9-hGem(1/110) was a gift from Giulio Draetta (Addgene plasmid # 71707 ; http://n2t.net/addgene:71707 ; RRID:Addgene_71707) |
| 参考文献部分: | Post-translational Regulation of Cas9 during G1 Enhances Homology-Directed Repair. Gutschner T, Haemmerle M, Genovese G, Draetta GF, Chin L. Cell Rep. 2016 Feb 3. pii: S2211-1247(16)00040-1. doi: 10.1016/j.celrep.2016.01.019. 10.1016/j.celrep.2016.01.019 PubMed 26854237 |
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pX330-U6-Chimeric_BB-CBh-hSpCas9-hGem(1/110)载体质粒基本信息
pX330-U6-Chimeric_BB-CBh-hSpCas9-hGem(1/110)质粒图谱载体图谱和pX330-U6-Chimeric_BB-CBh-hSpCas9-hGem(1/110)载体序列质粒序列多克隆位点信息
质粒图谱.png)
pX330-U6-Chimeric_BB-CBh-hSpCas9-hGem(1/110)质粒载体简介
pX330-U6-Chimeric_BB-CBh-hSpCas9-hGem(1/110)质粒序列载体序列
LOCUS pX330-U6-Chimeric_BB-CBh-hSpCas9-hGem(1/110) 8854 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION A human codon-optimized SpCas9 fused to first 110 amino acids of
human GEMININ and chimeric guide RNA expression plasmid.
ACCESSION .
VERSION .
KEYWORDS pX330-U6-Chimeric_BB-CBh-hSpCas9-hGem(1/110)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8854)
AUTHORS Gutschner T, Haemmerle M, Genovese G, Draetta GF, Chin L
TITLE Post-translational Regulation of Cas9 during G1 Enhances
Homology-Directed Repair.
JOURNAL Cell Rep. 2016 Feb 3. pii: S2211-1247(16)00040-1. doi:
10.1016/j.celrep.2016.01.019.
PUBMED 26854237
REFERENCE 2 (bases 1 to 8854)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8854
/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..1233
/label=hybrid intron
/note="hybrid between chicken beta-actin (CBA) and minute
virus of mice (MMV) introns (Gray et al., 2011)"
regulatory 1245..1254
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1254..1319
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 1326..1346
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1371..5471
/codon_start=1
/product="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
/label=Cas9
/note="generates RNA-guided double strand breaks in DNA"
/translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 5472..5519
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
primer_bind complement(5895..5912)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5901..6108
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 6117..6257
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 6117..6246
/label=AAV2 ITR
rep_origin 6332..6787
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6419..6438)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6629..6650
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(6804..6823)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6923..6945
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(6983..7001)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 7069..7173
/gene="bla"
/label=AmpR promoter
CDS 7174..8034
/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(7392..7411)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 8205..8793
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8694..8713
/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 ggggcgaggg gcggggcggg gcgaggcgga
901 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc
961 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgacgc
1021 tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg
1081 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag
1141 ctgagcaaga ggtaagggtt taagggatgg ttggttggtg gggtattaat gtttaattac
1201 ctggagcacc tgcctgaaat cacttttttt caggttggac cggtgccacc atggactata
1261 aggaccacga cggagactac aaggatcatg atattgatta caaagacgat gacgataaga
1321 tggccccaaa gaagaagcgg aaggtcggta tccacggagt cccagcagcc gacaagaagt
1381 acagcatcgg cctggacatc ggcaccaact ctgtgggctg ggccgtgatc accgacgagt
1441 acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga
1501 agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc acccggctga
1561 agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat ctgcaagaga
1621 tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg gaagagtcct
1681 tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac atcgtggacg
1741 aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa ctggtggaca
1801 gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg atcaagttcc
1861 ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg gacaagctgt
1921 tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc aacgccagcg
1981 gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc
2041 tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg attgccctga
2101 gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat gccaaactgc
2161 agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag atcggcgacc
2221 agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg ctgagcgaca
2281 tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg atcaagagat
2341 acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg
2401 agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc tacattgacg
2461 gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa aagatggacg
2521 gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag cagcggacct
2581 tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc attctgcggc
2641 ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag aagatcctga
2701 ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga ttcgcctgga
2761 tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg gtggacaagg
2821 gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac ctgcccaacg
2881 agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat aacgagctga
2941 ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc ggcgagcaga
3001 aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga
3061 aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag
3121 atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc aaggacaagg
3181 acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg accctgacac
3241 tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg
3301 acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg ctgagccgga
3361 agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat ttcctgaagt
3421 ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc ctgaccttta
3481 aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac gagcacattg
3541 ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg aaggtggtgg
3601 acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc gaaatggcca
3661 gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg aagcggatcg
3721 aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc
3781 agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat atgtacgtgg
3841 accaggaact ggacatcaac cggctgtccg actacgatgt ggaccatatc gtgcctcaga
3901 gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac aagaaccggg
3961 gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac tactggcggc
4021 agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc aaggccgaga
4081 gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg gtggaaaccc
4141 ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact aagtacgacg
4201 agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg
4261 atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac caccacgccc
4321 acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg
4381 aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg atcgccaaga
4441 gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac atcatgaact
4501 ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct ctgatcgaga
4561 caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc accgtgcgga
4621 aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct
4681 tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc agaaagaagg
4741 actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat tctgtgctgg
4801 tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg
4861 ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt ctggaagcca
4921 agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac tccctgttcg
4981 agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag aagggaaacg
5041 aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac tatgagaagc
5101 tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag cacaagcact
5161 acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc ctggccgacg
5221 ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc atcagagagc
5281 aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct gccgccttca
5341 agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag gtgctggacg
5401 ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac ctgtctcagc
5461 tgggaggcga caaaaggccg gcggccacga aaaaggccgg ccaggcaaaa aagaaaaagg
5521 gatccaagct tatgaatccc agtatgaagc agaaacaaga agaaatcaaa gagaatataa
5581 agaatagttc tgtcccaaga agaactctga agatgattca gccttctgca tctggatctc
5641 ttgttggaag agaaaatgag ctgtccgcag gcttgtccaa aaggaaacat cggaatgacc
5701 acttaacatc tacaacttcc agccctgggg ttattgtccc agaatctagt gaaaataaaa
5761 atcttggagg agtcacccag gagtcatttg atcttatgat taaagaaaat ccatcctctc
5821 agtattggaa ggaagtggca gaaaaacgga gaaaggcgct gtaactcgag gaattcctag
5881 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
5941 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
6001 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
6061 ggacagcaag ggggaggatt gggaagagaa tagcaggcat gctggggagc ggccgcagga
6121 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
6181 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
6241 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
6301 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
6361 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
6421 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
6481 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
6541 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
6601 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
6661 aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg
6721 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
6781 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
6841 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
6901 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
6961 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
7021 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
7081 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
7141 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
7201 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
7261 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
7321 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
7381 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
7441 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
7501 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
7561 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
7621 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
7681 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
7741 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
7801 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
7861 gctgataaat ctggagccgg tgagcgtgga agccgcggta tcattgcagc actggggcca
7921 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
7981 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
8041 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
8101 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
8161 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
8221 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
8281 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
8341 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
8401 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
8461 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
8521 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
8581 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
8641 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
8701 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
8761 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
8821 ttcctggcct tttgctggcc ttttgctcac atgt
//
