| 质粒名称: | eSpCas9(1.1) |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 71814 |
| 存储实验室: | Feng Zhang |
|
载体骨架基本信息 |
|
| 载体名称: | pX330-U6-Chimeric_BB-CBh-hSpCas9 |
| 载体出品公司: | Feng Zhang lab |
| 空载体大小: | 8505bp |
| 完整质粒大小: | 8505bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | Stbl3 |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | enhanced specificity Cas9 (1.1)/eSpCas9(1.1) |
| 物种来源: | S. pyogenes |
| 基因突变: | K848A, K1003A, & R1060A |
| GenBank编号 | JN247409 |
| 启动子: | CBh |
| 标签/融合蛋白: | 3xFLAG (N terminal on insert) |
| 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 | agggatggttggttggtggg |
| 3’测序引物 | CCAATCCTCCCCCTTGCTGT |
|
文献引用方法 |
|
| 材料和方法部分: | eSpCas9(1.1) was a gift from Feng Zhang (Addgene plasmid # 71814 ; http://n2t.net/addgene:71814 ; RRID:Addgene_71814) |
| 参考文献部分: | Rationally engineered Cas9 nucleases with improved specificity. Slaymaker IM, Gao L, Zetsche B, Scott DA, Yan WX, Zhang F. Science. 2015 Dec 1. pii: aad5227. 10.1126/science.aad5227 PubMed 26628643 |
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 eSpCas9(1.1) 8505 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses high specificity SpCas9. Px330-like plasmid..
ACCESSION .
VERSION .
KEYWORDS eSpCas9(1.1)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8505)
AUTHORS Slaymaker IM, Gao L, Zetsche B, Scott DA, Yan WX, Zhang F
TITLE Rationally engineered Cas9 nucleases with improved specificity.
JOURNAL Science. 2015 Dec 1. pii: aad5227.
PUBMED 26628643
REFERENCE 2 (bases 1 to 8505)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8505
/organism="synthetic DNA construct"
/mol_type="other DNA"
intron 172..399
/label=hybrid intron
/note="hybrid between chicken beta-actin (CBA) and minute
virus of mice (MMV) introns (Gray et al., 2011)"
regulatory 411..420
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 420..485
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 492..512
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 537..4637
/codon_start=1
/product="Cas9 endonuclease from the Streptococcus pyogenes
Type II CRISPR/Cas system, mutated to improve targeting
specificity (Slaymaker et al., 2016)"
/label=eSpCas9(1.1)
/note="carries the mutations K848A, K1003A, and R1060A"
/translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLADDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPALESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKAPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 4638..4685
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
primer_bind complement(4713..4730)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 4719..4926
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 4935..5075
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 4935..5064
/label=AAV2 ITR
rep_origin 5150..5605
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5237..5256)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5447..5468
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(5622..5641)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5741..5763
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(5801..5819)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 5887..5991
/gene="bla"
/label=AmpR promoter
CDS 5992..6852
/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(6210..6229)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7023..7611
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7512..7531
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 7673..7913
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 7673..7693
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 7844..7863
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 7940..8015
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
enhancer 8112..8397
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer;
contains an 18-bp deletion relative to the standard CMV
enhancer"
promoter 8399..171
/label=chicken beta-actin promoter
ORIGIN
1 gggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg gggcggggcg
61 aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg
121 gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct
181 gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc
241 tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt
301 aattagctga gcaagaggta agggtttaag ggatggttgg ttggtggggt attaatgttt
361 aattacctgg agcacctgcc tgaaatcact ttttttcagg ttggaccggt gccaccatgg
421 actataagga ccacgacgga gactacaagg atcatgatat tgattacaaa gacgatgacg
481 ataagatggc cccaaagaag aagcggaagg tcggtatcca cggagtccca gcagccgaca
541 agaagtacag catcggcctg gacatcggca ccaactctgt gggctgggcc gtgatcaccg
601 acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca
661 tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc
721 ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc
781 aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag
841 agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg
901 tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg
961 tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca
1021 agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca
1081 agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg
1141 ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg
1201 aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg
1261 ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca
1321 aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg
1381 gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga
1441 gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca
1501 agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc
1561 tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca
1621 ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga
1681 tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc
1741 ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg cacgccattc
1801 tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga
1861 tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg
1921 cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg
1981 acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc
2041 ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg
2101 agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg
2161 agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc
2221 agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg
2281 tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg
2341 acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc
2401 tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt
2461 tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga
2521 gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc
2581 tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga
2641 cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc
2701 acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg
2761 tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa
2821 tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc
2881 ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa
2941 acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt
3001 acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc
3061 ctcagagctt tctggccgac gactccatcg acaacaaggt gctgaccaga agcgacaaga
3121 accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact
3181 ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg
3241 ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg
3301 aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt
3361 acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg
3421 tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc
3481 acgcccacga cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtaccctg
3541 cgctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg
3601 ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca
3661 tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag gcgcctctga
3721 tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg
3781 tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag
3841 gcggcttcag caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa
3901 agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg
3961 tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc
4021 tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg
4081 aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc
4141 tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg
4201 gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg
4261 agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca
4321 agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg
4381 ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca
4441 gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg
4501 ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc
4561 tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt
4621 ctcagctggg aggcgacaaa aggccggcgg ccacgaaaaa ggccggccag gcaaaaaaga
4681 aaaagtaaga attcctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
4741 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
4801 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
4861 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagagaata gcaggcatgc
4921 tggggagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
4981 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
5041 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
5101 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
5161 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
5221 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
5281 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
5341 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
5401 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
5461 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg
5521 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
5581 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
5641 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
5701 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
5761 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
5821 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
5881 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
5941 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
6001 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
6061 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
6121 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
6181 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
6241 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
6301 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
6361 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
6421 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
6481 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
6541 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
6601 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
6661 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtggaag ccgcggtatc
6721 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
6781 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
6841 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
6901 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
6961 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
7021 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
7081 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
7141 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
7201 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
7261 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
7321 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
7381 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
7441 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
7501 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
7561 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
7621 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gtgagggcct
7681 atttcccatg attccttcat atttgcatat acgatacaag gctgttagag agataattgg
7741 aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta gaaagtaata
7801 atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc atatgcttac
7861 cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag gacgaaacac
7921 cgggtcttcg agaagacctg ttttagagct agaaatagca agttaaaata aggctagtcc
7981 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tgttttagag ctagaaatag
8041 caagttaaaa taaggctagt ccgtttttag cgcgtgcgcc aattctgcag acaaatggct
8101 ctagaggtac ccgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
8161 ccccgcccat tgacgtcaat agtaacgcca atagggactt tccattgacg tcaatgggtg
8221 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
8281 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attgtgccca gtacatgacc
8341 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc
8401 gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt
8461 ttgtatttat ttatttttta attattttgt gcagcgatgg gggcg
//
