| 质粒名称: | pEPSA5Cas9syn2 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 206963 |
| 存储实验室: | Daijie Chen |
| 体骨架基本信息 | |
| 载体名称: | pEPSA5 |
| 载体出品公司: | Clontech |
| 空载体大小: | 5500bp |
| 完整质粒大小: | 10062bp |
| 载体类型: | Bacterial Expression, CRISPR, Synthetic Biology |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 克隆方法: | Ligation Independent Cloning |
|
文献引用方法 |
|
| 材料和方法部分: | pEPSA5Cas9syn2 was a gift from Daijie Chen (Addgene plasmid # 206963 ; http://n2t.net/addgene:206963 ; RRID:Addgene_206963) |
| 参考文献部分: | Efficient Genome Editing in Most Staphylococcus aureus by Using the Restriction-Modification System Silent CRISPR-Cas9 Toolkit. Yang P, Yang J, Lin T, Liu Q, Yin Y, Chen D, Yang S. ACS Synth Biol. 2023 Oct 13. doi: 10.1021/acssynbio.3c00339. 10.1021/acssynbio.3c00339 PubMed 37830328 |
买家导航
pEPSA5Cas9syn2载体质粒基本信息
pEPSA5Cas9syn2质粒图谱载体图谱和pEPSA5Cas9syn2载体序列质粒序列多克隆位点信息
pEPSA5Cas9syn2质粒载体简介
pEPSA5Cas9syn2质粒序列载体序列
LOCUS pEPSA5Cas9syn2 10062 bp ds-DNA circular SYN 10-NOV-2023
DEFINITION S. aureus genome editing vector removed these two motifs
AGGNNNNNGAT, CCAYNNNNNNTGT, CCAYNNNNNNGTA, GGANNNNNNNNTCG, and
GWAGNNNNNNGAT.
ACCESSION .
VERSION .
KEYWORDS pEPSA5Cas9syn2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10062)
TITLE Efficient Genome Editing in Most Staphylococcus aureus by Using the
Restriction-modification Systems Silent CRISPR-Cas9 Toolkit
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 10062)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Nov 10, 2023 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..10062
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin complement(260..805)
/direction=LEFT
/label=p15A ori
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin.
"
primer_bind complement(340..359)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
misc_RNA 4102..4177
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
CDS 4418..8524
/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"
primer_bind complement(8555..8572)
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
terminator 8725..8811
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 8903..8930
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
CDS complement(9072..9932)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 9695..9714
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(9933..10037)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ggcggccgca ctggcttact atgttggcac tgatgagggt gtcagtgaag tgcttcatgt
61 ggcaggagaa aaaaggctgc accggtgcgt cagcagaata tgtgatacag gatatattcc
121 gcttcctcgc tcactgactc gctacgctcg gtcgttcgac tgcggcgagc ggaaatggct
181 tacgaacggg gcggagattt cctggaagat gccaggaaga tacttaacag ggaagtgaga
241 gggccgcggc aaagccgttt ttccataggc tccgcccccc tgacaagcat cacgaaatct
301 gacgctcaaa tcagtggtgg cgaaacccga caggactata aagataccag gcgtttcccc
361 ctggcggctc cctcgtgcgc tctcctgttc ctgcctttcg gtttaccggt gtcattccgc
421 tgttatggcc gcgtttgtct cattccacgc ctgacactca gttccgggta ggcagttcgc
481 tccaagctgg actgtatgca cgaacccccc gttcagtccg accgctgcgc cttatccggt
541 aactatcgtc ttgagtccaa cccggaaaga catgcaaaag caccactggc agcagccact
601 ggtaattgat ttagaggagt tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag
661 gacaagtttt ggtgactgcg ctcctccaag ccagttacct cggttcaaag agttggtagc
721 tcagagaacc ttcgaaaaac cgccctgcaa ggcggttttt tcgttttcag agcaagagat
781 tacgcgcaga ccaaaacgat ctcaagaaga tcatcttatg cggccgcttc tttcctgcgt
841 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
901 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
961 ggtattttct ccttacgcat ctgtgcggta tttcacaccg cataggtaga tccctcgacc
1021 tgcaggcatg caagcttctg taggttttta ggcataaaac tatatgattt acccctaaat
1081 ctttaaaatg ccccttaaaa ttcaaaataa aggcatttaa aatttaaata tttcttgtga
1141 taaagtttgt taaaaaggag tggttttatg actgttatgt ggttatcgat tataggtatg
1201 tggttttgta ttggaatggc attttttgct atcaaggtta ttaaaaataa aaattagacc
1261 acgcatttat gccgagaaaa tttattgtgc gttgagaaga acccttaact aaacttgcag
1321 acgaatgtcg gcatagcgtg agctattaag ccgaccattc gacaagtttt gggattgtta
1381 agggttccga ggctcaacgt caataaagca attggaataa agaagcgaaa aaggagaagt
1441 cggttcagaa aaagaagcat atgcatctgg agctgtaata taaaaacctt cttcaactaa
1501 cggggcaggt tagtgacatt agaaaaccga ctgtaaaaag tacagtcggc attatctcat
1561 attataaaag ccagtcatta ggcctatctg acaattcctg aatagagttc ataaacaatc
1621 ctgcatgata accatcacaa acagaatgat gtacctgtaa agatagcggt aaatatattg
1681 aattaccttt attaatgaat tttcctgctg taataatggg tagaaggtaa ttactattat
1741 tattgatatt taagttaaac ccagtaaatg aagtccatgg aataatagaa agagaaaaag
1801 cattttcagg tataggtgtt ttgggaaaca atttccccga accattatat ttctctacat
1861 cagaaaggta taaatcataa aactctttga agtcattctt tacaggagtc caaataccag
1921 agaatgtttt agatacacca tcaaaaattg tataaagtgg ctctaactta tcccaataac
1981 ctaactctcc gtcgctattg taaccagttc taaaagctgt atttgagttt atcacccttg
2041 tcactaagaa aataaatgca gggtaaaatt tatatccttc ttgttttatg tttcggtata
2101 aaacactaat atcaatttct gtggttatac taaaagtcgt ttgttggttc aaataatgat
2161 taaatatctc ttttctcttc caattgtcta aatcaatttt attaaagttc atttgatatg
2221 cctcctaaat ttttatctaa agtgaattta ggaggcttac ttgtctgctt tcttcattag
2281 aatcaatcct tttttaaaag tcaatattac tgtaacataa atatatattt taaaaatatc
2341 ccactttatc caattttcgt ttgttgaact aatgggtgct ttagttgaag aataaaagac
2401 cacattaaaa aatgtggtct tttgtgtttt tttaaaggat ttgagcgtag cgaaaaatcc
2461 ttttctttct tatcttgata ataagggtaa ctattgccgg cgaggctagt tacccttaag
2521 ttattggtat gactggtttt aagcgcaaaa aaagttgctt tttcgtacct attaatgtat
2581 cgttttaaat gaatagtaaa aaacatacat agaaagggga aaaagcaact ttttttattg
2641 tcatagtttg tgaaaactaa gttgttttta tgtgttataa catggaaaag tatactgaga
2701 aaaaacaaag aaatcaagta tttcagaaat ttattaaacg tcatattgga gagaatcaaa
2761 tggatttagt tgaagattgc aatacatttc tgtcttttgt agctgataaa actttagaaa
2821 aacagaaatt atataaagct aattcttgta aaaatcgatt ttgtcctgtc tgtgcttgga
2881 gaaaagctag aaaagatgca ttgggtttat ctttgatgat gcaatatatt aagcagcaag
2941 agaaaaagga gtttatcttt ttaactttga ctacacctaa tgtaatgagt gatgaattag
3001 aaaatgaaat aaaacgttat aataattctt ttagaaaact tataaagaga aaaaaagtag
3061 gtagtgttat aaagggatat gttcgtaagt tagagattac atataataaa aaaagagatg
3121 attataatcc tcattttcat gtgttaattg cagtaaataa atcgtatttc acagataaaa
3181 gatattatat tagccaacaa gaatggttag atttatggcg tgatgtaacg ggcatttcag
3241 aaataacaca agttcaagtt caaaaaataa gacaaaataa taataaagaa ttatatgaaa
3301 tggctaagta ttctggtaaa gatagtgatt atttaataaa tcaaaaagtc tttgatgcat
3361 tttataaatc acttaaaggt aaacaggtat tagtttattc aggattattt aaagaggcta
3421 aaaagaaatt aaaaaatggg gatttagatt acttaaaaga aattgatcca accgaatata
3481 tctatcaaat tttttatatt tggaaacaaa aagagtattt agctagtgaa ctttatgact
3541 taacagaaca agaaaaaaga gaaattaatc acaaaatgat agacgaaatc gaggaagaac
3601 aataacaaaa tataagtgct aacagtcgtc tgcaagttta gttaagggtt cttctcaacg
3661 cacaataaat tttctcggca taaatgcgtg gtctaatttt tatttttaat aaccttgata
3721 gcaaaaaatg ccattccaat acaaaaccac atacctataa tcgataacca cataacagtc
3781 ataaaaccac tcctttttaa caaactttat cacaagaaat attttggcat tctacgacta
3841 taacttaaat ttatattttt tactttataa tatataattg attatagaat aatgttgctc
3901 atatcgtttg ccaacatcta gtactcaaat tacactatgt tacacttggt aatattaacc
3961 gaacttcccc tgtccaaatt agtttttacg gttcctggcc ttttgctggc cttttgctca
4021 catgttcttt cctgcgttgt cgacagagtt tgcaaaatat acaggggatt atatataatg
4081 gaaacgagac cattggtctc agttttagag ctagaaatag caagttaaaa taaggctagt
4141 ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt ttttgagatc tgtccatacc
4201 catggtctag aatgctcgag tcagaaaaat atacctgtat cttttttcaa aagcaaacat
4261 gctttgggta aacatgtagg tattaacgtc aatgcgacaa tagtagcatt gattaaatga
4321 gaattagtaa gtgttttact tactaaattt tatttaacct aaaaatgaac cacctggatg
4381 tgtgggatta aaaagtgaag agaggaggac gtatcacatg gataagaaat actcaatagg
4441 cttagatatc ggcacaaata gcgtcggatg ggcggtgatc actgatgaat ataaggttcc
4501 gtctaaaaag ttcaaggttc tgggaaatac agaccgccac agtatcaaaa aaaatcttat
4561 aggggctctt ttatttgaca gtggagagac agcggaagcg actcgtctca aacggacagc
4621 tcgtagaagg tatacacgtc ggaagaatcg tatttgttat ctacaggaga ttttttcaaa
4681 tgagatggcg aaagtagatg atagtttctt tcatcgactt gaagagtctt ttttggtgga
4741 agaagacaag aagcatgaac gtcatcctat ttttggaaat atagtagatg aagttgctta
4801 tcatgagaaa tatccaacta tctatcatct gcgaaaaaaa ttggtagatt ctactgataa
4861 agcggatttg cgcttaatct atttggcctt agcgcatatg attaagtttc gtggtcattt
4921 tttgattgag ggagatttaa atcctgataa tagtgatgtg gacaaactat ttatccagtt
4981 ggtacaaacc tacaatcaat tatttgaaga aaaccctatt aacgcaagtg gagtagatgc
5041 taaagcgatt ctttctgcac gattgagtaa atcaagacga ttagaaaatc tcattgctca
5101 gctccccggt gagaagaaaa atggcttatt tgggaatctc attgctttgt cattgggttt
5161 gacccctaat tttaaatcaa attttgattt ggcagaagat gctaaattac agctttcaaa
5221 agatacttac gatgatgatt tagataattt attggcgcaa attggagatc aatatgctga
5281 tttgtttttg gcagctaaga atttatcaga tgctatttta ctttcagata tcctaagagt
5341 aaatactgaa ataactaagg ctcccctatc agcttcaatg attaaacgct acgatgaaca
5401 tcatcaagac ttgactcttt taaaagcttt agttcgacaa caacttccag aaaagtataa
5461 agaaatcttt tttgatcaat caaaaaacgg atatgcaggt tatattgatg ggggagctag
5521 ccaagaagaa ttttataaat ttatcaaacc aattttagaa aaaatggatg gtactgagga
5581 attattggtg aaactaaatc gtgaagattt gctgcgcaag caacggacct ttgacaacgg
5641 ctctattccc catcaaattc acttgggtga gctgcatgct attttgagaa gacaagaaga
5701 cttttatcca tttttaaaag acaatcgtga gaagattgaa aaaatcttga cttttcgaat
5761 tccttattat gttggtccat tggcgcgtgg caatagtcgt tttgcatgga tgactcggaa
5821 gtctgaagaa acaattaccc catggaattt tgaagaggtt gtcgataaag gtgcttcagc
5881 tcaatcattt attgaacgca tgacaaactt tgataaaaat cttccaaatg aaaaagtact
5941 accaaaacat agtttgcttt atgagtattt tacggtttat aacgaattga caaaggtcaa
6001 atatgttact gaaggaatgc gaaaaccagc atttctttca ggtgaacaga agaaagccat
6061 tgttgattta ctcttcaaaa caaatcgaaa agtaaccgtt aagcaattaa aagaagatta
6121 tttcaaaaaa atagaatgtt ttgatagtgt tgaaatttca ggagttgaag atagatttaa
6181 tgcttcatta ggtacctacc atgatttgct aaaaattatt aaagataaag attttttgga
6241 taatgaagaa aatgaagata tcttagagga tattgtttta acattgacct tatttgaaga
6301 tagagagatg attgaggaaa gacttaaaac atatgctcac ctctttgatg ataaggtgat
6361 gaaacagctt aaacgtcgcc gttatactgg ttggggacgt ttgtctcgaa aattgattaa
6421 tggtattagg gataagcaat ctggcaaaac aatattagat tttttgaaat cagatggttt
6481 tgccaatcgc aattttatgc agctgatcca tgatgatagt ttgacattta aagaagacat
6541 tcaaaaagca caagtgtctg gacaaggcga tagtttacat gaacatattg caaatttagc
6601 tggtagccct gctattaaaa aaggtatttt acagactgta aaagttgttg atgaattggt
6661 caaagtaatg gggcggcata agccagaaaa tatcgttatt gaaatggcac gtgaaaatca
6721 gacaactcaa aagggccaga aaaattcgcg agagcgtatg aaacgaatcg aagaaggtat
6781 caaagaatta ggaagtcaga ttcttaaaga gcatcctgtt gaaaatactc aattgcaaaa
6841 tgaaaagctc tatctctatt atctccaaaa tggaagagac atgtatgtgg accaagaatt
6901 agatattaat cgtttaagtg attatgatgt cgatcacatt gttccacaaa gtttccttaa
6961 agacgattca atagacaata aggtcttaac gcgttctgat aaaaatcgtg gtaaatcgga
7021 taacgttcca agtgaagaag tagtcaaaaa gatgaaaaac tattggagac aacttctaaa
7081 cgccaagtta atcactcaac gtaagtttga taatttaacg aaagctgaac gtggaggttt
7141 gagtgaactt gataaagctg gttttatcaa acgccaattg gttgaaactc gccaaatcac
7201 taagcatgtg gcacaaattt tggatagtcg catgaatact aaatacgatg aaaatgataa
7261 acttattcga gaggttaaag tgattacctt aaaatctaaa ttagtttctg acttccgaaa
7321 agattttcaa ttctataaag tacgtgagat taacaattac catcatgccc atgatgcgta
7381 tctaaatgcc gtcgttggaa ctgctttgat taagaaatat ccaaaacttg aatcggagtt
7441 tgtctatggt gattataaag tttatgatgt tcgtaaaatg attgctaagt ctgagcaaga
7501 aataggcaaa gcaaccgcaa aatatttctt ttactctaat atcatgaact ttttcaaaac
7561 agaaattaca cttgcaaatg gagagattcg caaacgccct ctaatcgaaa ctaatgggga
7621 aactggagaa attgtctggg ataaagggcg agattttgcc acagtgcgca aagtattgtc
7681 catgccccaa gtcaatattg tcaagaaaac agaagtacag acaggcggat tctccaagga
7741 gtcaatttta ccaaaaagaa attcggacaa gcttattgct cgtaaaaaag actgggatcc
7801 aaaaaaatat ggtggttttg atagtccaac ggtagcttat tcagtcctag tggttgctaa
7861 ggtggaaaaa gggaaatcga agaagttaaa atccgttaaa gagttactag ggatcacaat
7921 tatggaaaga agttcctttg aaaaaaatcc gattgacttt ttagaagcta aaggatataa
7981 ggaagttaaa aaagacttaa tcattaaact acctaaatat agtctttttg agttagaaaa
8041 cggtcgtaaa cggatgctgg ctagtgccgg agaattacaa aaaggaaatg agctggctct
8101 gccaagcaaa tatgtgaatt ttttatattt agctagtcat tatgaaaagt tgaagggtag
8161 tccagaagat aacgaacaaa aacaattgtt tgtggagcag cataagcatt atttagatga
8221 gattattgag caaatcagtg aattttctaa gcgtgttatt ttagcagatg ccaatttaga
8281 taaagttctt agtgcatata acaaacatag agacaaacca atacgtgaac aagcagaaaa
8341 tattattcat ttatttacgt tgacgaatct tggagctccc gctgctttta aatattttga
8401 tacaacaatt gatcgtaaac gatatacgtc tacaaaagag gttttagatg ccactcttat
8461 ccatcaatcc atcactggtc tttatgaaac acgcattgat ttgagtcagc taggaggtga
8521 ctaatcgacc tgcagccaag cttgggcttt tcagcctgat acagattaaa tcagaacgca
8581 gaagcggtct gataaaacag aatttgcctg gcggcagtag cgcggtggtc ccacctgacc
8641 ccatgccgaa ctcagaagtg aaacgccgta gcgccgatgg tagtgtggcg tctccccttg
8701 cgagagtagg gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc
8761 tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga
8821 gcggatttga acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa
8881 actgccaggc atcaaattaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta
8941 caaactcttt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgcccatcgt
9001 atcgatgata agctgtcaaa catgagaatt aaatcaatct aaagtatata tgagtaaact
9061 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
9121 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
9181 ccatctggcc ccagtgctgc aatgataccg cgtgacccac gctcaccggc tccagattta
9241 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
9301 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
9361 agtttgcgca acgttgttgc cattgctact ggcatcgtag tgtcacgctc gtcgtttggt
9421 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
9481 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
9541 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
9601 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
9661 cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact
9721 ttaaaagtgc tcatcattgg aaaacgctct tcggggcgaa aactctcaag gatcttaccg
9781 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
9841 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
9901 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
9961 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
10021 caaatagggg ttccgcgcac atttccccga aaagtgccac ct
//
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pEPSA5Cas9syn2质粒载体应用举例
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