| 质粒名称: | pET-28b-Cas9-His |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 47327 |
| 存储实验室: | Alex Schier |
|
载体骨架基本信息 |
|
| 载体名称: | pN1 |
| 载体出品公司: | Clontech |
| 空载体大小: | 5368bp |
| 完整质粒大小: | 9375bp |
| 载体类型: | 细菌细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Cas9 |
| 物种来源: | 光合细菌 Rhodopseudomonas palustris |
| 插入大小: | 4100 bp |
| 标签/融合蛋白: | His (C terminal on backbone) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | NcoI(未破坏) |
| 3’克隆位点: | NotI (未破坏) |
| 5’测序引物 | T7 |
|
文献引用方法 |
|
| 材料和方法部分: | pET-28b-Cas9-His was a gift from Alex Schier (Addgene plasmid # 47327 ; http://n2t.net/addgene:47327 ; RRID:Addgene_47327) |
| 参考文献部分: | Efficient mutagenesis by Cas9 protein-mediated oligonucleotide insertion and large-scale assessment of single-guide RNAs. Gagnon JA, Valen E, Thyme SB, Huang P, Ahkmetova L, Pauli A, Montague TG, Zimmerman S, Richter C, Schier AF. PLoS One. 2014 May 29;9(5):e98186. doi: 10.1371/journal.pone.0098186. eCollection 2014. PONE-D-14-14210 [pii] PubMed 24873830 |
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
载体简介
载体序列
LOCUS pET-28b-Cas9-His 9375 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION For in vitro expression and purification of Cas9 protein.
ACCESSION .
VERSION .
KEYWORDS pET-28b-Cas9-His
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9375)
AUTHORS Gagnon JA, Valen E, Thyme SB, Huang P, Ahkmetova L, Pauli A,
Montague TG, Zimmerman S, Richter C, Schier AF
TITLE Efficient mutagenesis by Cas9 protein-mediated oligonucleotide
insertion and large-scale assessment of single-guide RNAs.
JOURNAL PLoS One. 2014 May 29;9(5):e98186. doi:
10.1371/journal.pone.0098186. eCollection 2014.
PUBMED 24873830
REFERENCE 2 (bases 1 to 9375)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..9375
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 5..144
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(70..89)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 230..252
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(246..437)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(1246..2328)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind 2290..2309
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(2329..2406)
/gene="lacI"
/label=lacI promoter
primer_bind 2612..2631
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 2715..2734
/label=T7
/note="T7 promoter, forward primer"
promoter 2715..2733
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 2734..2758
/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)."
RBS 2773..2795
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 2802..6905
/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="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
CDS 6918..6938
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 6954..6971
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(7024..7042)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 7038..7085
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 7122..7577
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(7209..7228)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 7419..7440
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(7669..8484)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 8392..8411
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 8476..8495
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 8606..9194
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 9095..9114
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 9348..9365
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg
61 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
121 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
181 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
241 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca
301 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg
361 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt
421 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg
481 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
541 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
601 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc
661 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt
721 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac
781 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca
841 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc
901 cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga
961 ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg
1021 atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc
1081 acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg
1141 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
1201 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
1261 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
1321 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
1381 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
1441 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
1501 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
1561 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
1621 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
1681 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
1741 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
1801 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
1861 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
1921 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
1981 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
2041 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
2101 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
2161 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
2221 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
2281 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
2341 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
2401 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta
2461 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc
2521 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
2581 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
2641 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct
2701 cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata acaattcccc
2761 tctagaaata attttgttta actttaagaa ggagatatac catggacaag aagtactcca
2821 ttgggctcga tatcggcaca aacagcgtcg gctgggccgt cattacggac gagtacaagg
2881 tgccgagcaa aaaattcaaa gttctgggca ataccgatcg ccacagcata aagaagaacc
2941 tcattggcgc cctcctgttc gactccgggg agacggccga agccacgcgg ctcaaaagaa
3001 cagcacggcg cagatatacc cgcagaaaga atcggatctg ctacctgcag gagatcttta
3061 gtaatgagat ggctaaggtg gatgactctt tcttccatag gctggaggag tcctttttgg
3121 tggaggagga taaaaagcac gagcgccacc caatctttgg caatatcgtg gacgaggtgg
3181 cgtaccatga aaagtaccca accatatatc atctgaggaa gaagcttgta gacagtactg
3241 ataaggctga cttgcggttg atctatctcg cgctggcgca tatgatcaaa tttcggggac
3301 acttcctcat cgagggggac ctgaacccag acaacagcga tgtcgacaaa ctctttatcc
3361 aactggttca gacttacaat cagcttttcg aagagaaccc gatcaacgca tccggagttg
3421 acgccaaagc aatcctgagc gctaggctgt ccaaatcccg gcggctcgaa aacctcatcg
3481 cacagctccc tggggagaag aagaacggcc tgtttggtaa tcttatcgcc ctgtcactcg
3541 ggctgacccc caactttaaa tctaacttcg acctggccga agatgccaag cttcaactga
3601 gcaaagacac ctacgatgat gatctcgaca atctgctggc ccagatcggc gaccagtacg
3661 cagacctttt tttggcggca aagaacctgt cagacgccat tctgctgagt gatattctgc
3721 gagtgaacac ggagatcacc aaagctccgc tgagcgctag tatgatcaag cgctatgatg
3781 agcaccacca agacttgact ttgctgaagg cccttgtcag acagcaactg cctgagaagt
3841 acaaggaaat tttcttcgat cagtctaaaa atggctacgc cggatacatt gacggcggag
3901 caagccagga ggaattttac aaatttatta agcccatctt ggaaaaaatg gacggcaccg
3961 aggagctgct ggtaaagctt aacagagaag atctgttgcg caaacagcgc actttcgaca
4021 atggaagcat cccccaccag attcacctgg gcgaactgca cgctatcctc aggcggcaag
4081 aggatttcta cccctttttg aaagataaca gggaaaagat tgagaaaatc ctcacatttc
4141 ggatacccta ctatgtaggc cccctcgccc ggggaaattc cagattcgcg tggatgactc
4201 gcaaatcaga agagaccatc actccctgga acttcgagga agtcgtggat aagggggcct
4261 ctgcccagtc cttcatcgaa aggatgacta actttgataa aaatctgcct aacgaaaagg
4321 tgcttcctaa acactctctg ctgtacgagt acttcacagt ttataacgag ctcaccaagg
4381 tcaaatacgt cacagaaggg atgagaaagc cagcattcct gtctggagag cagaagaaag
4441 ctatcgtgga cctcctcttc aagacgaacc ggaaagttac cgtgaaacag ctcaaagaag
4501 actatttcaa aaagattgaa tgtttcgact ctgttgaaat cagcggagtg gaggatcgct
4561 tcaacgcatc cctgggaacg tatcacgatc tcctgaaaat cattaaagac aaggacttcc
4621 tggacaatga ggagaacgag gacattcttg aggacattgt cctcaccctt acgttgtttg
4681 aagataggga gatgattgaa gaacgcttga aaacttacgc tcatctcttc gacgacaaag
4741 tcatgaaaca gctcaagagg cgccgatata caggatgggg gcggctgtca agaaaactga
4801 tcaatgggat ccgagacaag cagagtggaa agacaatcct ggattttctt aagtccgatg
4861 gatttgccaa ccggaacttc atgcagttga tccatgatga ctctctcacc tttaaggagg
4921 acatccagaa agcacaagtt tctggccagg gggacagtct tcacgagcac atcgctaatc
4981 ttgcaggtag cccagctatc aaaaagggaa tactgcagac cgttaaggtc gtggatgaac
5041 tcgtcaaagt aatgggaagg cataagcccg agaatatcgt tatcgagatg gcccgagaga
5101 accaaactac ccagaaggga cagaagaaca gtagggaaag gatgaagagg attgaagagg
5161 gtataaaaga actggggtcc caaatcctta aggaacaccc agttgaaaac acccagcttc
5221 agaatgagaa gctctacctg tactacctgc agaacggcag ggacatgtac gtggatcagg
5281 aactggacat caatcggctc tccgactacg acgtggatca tatcgtgccc cagtcttttc
5341 tcaaagatga ttctattgat aataaagtgt tgacaagatc cgataaaaat agagggaaga
5401 gtgataacgt cccctcagaa gaagttgtca agaaaatgaa aaattattgg cggcagctgc
5461 tgaacgccaa actgatcaca caacggaagt tcgataatct gactaaggct gaacgaggtg
5521 gcctgtctga gttggataaa gccggcttca tcaaaaggca gcttgttgag acacgccaga
5581 tcaccaagca cgtggcccaa attctcgatt cacgcatgaa caccaagtac gatgaaaatg
5641 acaaactgat tcgagaggtg aaagttatta ctctgaagtc taagctggtc tcagatttca
5701 gaaaggactt tcagttttat aaggtgagag agatcaacaa ttaccaccat gcgcatgatg
5761 cctacctgaa tgcagtggta ggcactgcac ttatcaaaaa atatcccaag cttgaatctg
5821 aatttgttta cggagactat aaagtgtacg atgttaggaa aatgatcgca aagtctgagc
5881 aggaaatagg caaggccacc gctaagtact tcttttacag caatattatg aattttttca
5941 agaccgagat tacactggcc aatggagaga ttcggaagcg accacttatc gaaacaaacg
6001 gagaaacagg agaaatcgtg tgggacaagg gtagggattt cgcgacagtc cggaaggtcc
6061 tgtccatgcc gcaggtgaac atcgttaaaa agaccgaagt acagaccgga ggcttctcca
6121 aggaaagtat cctcccgaaa aggaacagcg acaagctgat cgcacgcaaa aaagattggg
6181 accccaagaa atacggcgga ttcgattctc ctacagtcgc ttacagtgta ctggttgtgg
6241 ccaaagtgga gaaagggaag tctaaaaaac tcaaaagcgt caaggaactg ctgggcatca
6301 caatcatgga gcgatcaagc ttcgaaaaaa accccatcga ctttctcgag gcgaaaggat
6361 ataaagaggt caaaaaagac ctcatcatta agcttcccaa gtactctctc tttgagcttg
6421 aaaacggccg gaaacgaatg ctcgctagtg cgggcgagct gcagaaaggt aacgagctgg
6481 cactgccctc taaatacgtt aatttcttgt atctggccag ccactatgaa aagctcaaag
6541 ggtctcccga agataatgag cagaagcagc tgttcgtgga acaacacaaa cactaccttg
6601 atgagatcat cgagcaaata agcgaattct ccaaaagagt gatcctcgcc gacgctaacc
6661 tcgataaggt gctttctgct tacaataagc acagggataa gcccatcagg gagcaggcag
6721 aaaacattat ccacttgttt actctgacca acttgggcgc gcctgcagcc ttcaagtact
6781 tcgacaccac catagacaga aagcggtaca cctctacaaa ggaggtcctg gacgccacac
6841 tgattcatca gtcaattacg gggctctatg aaacaagaat cgacctctct cagctcggtg
6901 gagacagcag ggctgacccc aagaagaaga ggaaggtggc ggccgcactc gagcaccacc
6961 accaccacca ctgagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
7021 ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
7081 ttttgctgaa aggaggaact atatccggat tggcgaatgg gacgcgccct gtagcggcgc
7141 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
7201 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
7261 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
7321 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
7381 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
7441 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
7501 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
7561 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
7621 ttatttttct aaatacattc aaatatgtat ccgctcatga attaattctt agaaaaactc
7681 atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg
7741 aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag
7801 atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc
7861 ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga
7921 gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc
7981 gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag
8041 acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg
8101 caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac
8161 ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg
8221 gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat
8281 ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc
8341 atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc
8401 ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc tagagcaaga
8461 cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag
8521 ttttattgtt catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
8581 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
8641 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
8701 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
8761 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
8821 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
8881 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
8941 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
9001 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
9061 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
9121 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
9181 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
9241 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
9301 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
9361 gcgaggaagc ggaag
//
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