| 质粒名称: | pCas9 |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 42876 |
| 存储实验室: | Luciano Marraffini |
|
载体骨架基本信息 |
|
| 载体名称: | pACYC184 |
| 空载体大小: | 4244 bp |
| 完整质粒大小: | 9326 bp |
| 载体类型: | 细菌细胞表达 |
| 载体抗性: | 氯霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | tracr/Cas9 |
|
文献引用方法 |
|
| 材料和方法部分: | pCas9 was a gift from Luciano Marraffini (Addgene plasmid # 42876 ; http://n2t.net/addgene:42876 ; RRID:Addgene_42876) |
| 参考文献部分: | RNA-guided editing of bacterial genomes using CRISPR-Cas systems. Jiang W, Bikard D, Cox D, Zhang F, Marraffini LA. Nat Biotechnol. 2013 Jan 29. doi: 10.1038/nbt.2508. 10.1038/n |
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 pCas9 9326 bp ds-DNA circular SYN 17-JUN-2022
DEFINITION Bacterial expression of Cas9 nuclease, tracrRNA and crRNA guide.
ACCESSION .
VERSION .
KEYWORDS pCas9
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9326)
AUTHORS Jiang W, Bikard D, Cox D, Zhang F, Marraffini LA
TITLE RNA-guided editing of bacterial genomes using CRISPR-Cas systems.
JOURNAL Nat Biotechnol. 2013 Jan 29. doi: 10.1038/nbt.2508.
PUBMED 23360965
REFERENCE 2 (bases 1 to 9326)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jun 17, 2022 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..9326
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 386..405
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 602..619
/label=L4440
/note="L4440 vector, forward primer"
promoter 1011..1113
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
CDS 1114..1773
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(1180..1199)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
primer_bind 3626..3645
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
repeat_region 4074..4109
/label=DR
/note="direct repeat for the Streptococcus pyogenes
CRISPR/Cas system"
repeat_region 4140..4175
/label=DR
/note="direct repeat for the Streptococcus pyogenes
CRISPR/Cas system"
misc_feature complement(4176..4307)
/label=crRNA leader
/note="crRNA leader sequence for the Streptococcus pyogenes
CRISPR/Cas system"
CDS complement(4328..8434)
/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"
misc_RNA 8730..8808
/label=tracrRNA
/note="trans-activating CRISPR RNA for the Streptococcus
pyogenes CRISPR/Cas9 system"
promoter complement(9126..9154)
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
rep_origin 9266..485
/direction=RIGHT
/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.
"
ORIGIN
1 ggcggttttt cgaaggttct ctgagctacc aactctttga accgaggtaa ctggcttgga
61 ggagcgcagt caccaaaact tgtcctttca gtttagcctt aaccggcgca tgacttcaag
121 actaactcct ctaaatcaat taccagtggc tgctgccagt ggtgcttttg catgtctttc
181 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcggact gaacgggggg
241 ttcgtgcata cagtccagct tggagcgaac tgcctacccg gaactgagtg tcaggcgtgg
301 aatgagacaa acgcggccat aacagcggaa tgacaccggt aaaccgaaag gcaggaacag
361 gagagcgcac gagggagccg ccagggggaa acgcctggta tctttatagt cctgtcgggt
421 ttcgccacca ctgatttgag cgtcagattt cgtgatgctt gtcagggggg cggagcctat
481 ggaaaaacgg ctttgccgcg gccctctcac ttccctgtta agtatcttcc tggcatcttc
541 caggaaatct ccgccccgtt cgtaagccat ttccgctcgc cgcagtcgaa cgaccgagcg
601 tagcgagtca gtgagcgagg aagcggaata tatcctgtat cacatattct gctgacgcac
661 cggtgcagcc ttttttctcc tgccacatga agcacttcac tgacaccctc atcagtgcca
721 acatagtaag ccagtataca ctccgctagc gctgatgtcc ggcggtgctt ttgccgttac
781 gcaccacccc gtcagtagct gaacaggagg gacagctgat agaaacagaa gccactggag
841 cacctcaaaa acaccatcat acactaaatc agtaagttgg cagcatcacc cgacgcactt
901 tgcgccgaat aaatacctgt gacggaagat cacttcgcag aataaataaa tcctggtgtc
961 cctgttgata ccgggaagcc ccgggccaac ttttggcgaa aatgagacgt tgatcggcac
1021 gtaagaggtt ccaactttca ccataatgaa ataagatcac taccgggcgt attttttgag
1081 ttatcgagat tttcaggagc taaggaagct aaaatggaga aaaaaatcac tggatatacc
1141 accgttgata tatcccaatg gcatcgtaaa gaacattttg aggcatttca gtcagttgct
1201 caatgtacct ataaccagac cgttcagctg gatattacgg cctttttaaa gaccgtaaag
1261 aaaaataagc acaagtttta tccggccttt attcacattc ttgcccgcct gatgaatgct
1321 catccggaat tccgtatggc aatgaaagac ggtgagctgg tgatatggga tagtgttcac
1381 ccttgttaca ccgttttcca tgagcaaact gaaacgtttt catcgctctg gagtgaatac
1441 cacgacgatt tccggcagtt tctacacata tattcgcaag atgtggcgtg ttacggtgaa
1501 aacctggcct atttccctaa agggtttatt gagaatatgt ttttcgtctc agccaatccc
1561 tgggtgagtt tcaccagttt tgatttaaac gtggccaata tggacaactt cttcgccccc
1621 gttttcacca tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt
1681 caggttcatc atgccgtttg tgatggcttc catgtcggca gaatgcttaa tgaattacaa
1741 cagtactgcg atgagtggca gggcggggcg taattttttt aaggcagtta ttggtgccct
1801 taaacgcctg gttgctacgc ctgaataagt gataataagc ggatgaatgg cagaaattcg
1861 aaagcaaatt cgacccggtc gtcggttcag ggcagggtcg ttaaatagcc gcttatgtct
1921 attgctggtt taccggttta ttgactaccg gaagcagtgt gaccgtgtgc ttctcaaatg
1981 cctgaggcca gtttgctcag gctctccccg tggaggtaat aattgacgat atgatcattt
2041 attctgcctc ccagagcctg ataaaaacgg ttagcgcttc gttaatacag atgtaggtgt
2101 tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc
2161 ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgctgccgg
2221 cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat
2281 gccccgcgcc caccggaagg agctaccggc agcggtgcgg actgttgtaa ctcagaataa
2341 gaaatgaggc cgctcatggc gttgactctc agtcatagta tcgtggtatc accggttggt
2401 tccactctct gttgcgggca acttcagcag cacgtagggg acttccgcgt ttccagactt
2461 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
2521 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
2581 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggccag
2641 gacccaacgc tgcccgagat gcgccgcgtg cggctgctgg agatggcgga cgcgatggat
2701 atgttctgcc aagggttggt ttgcgcattc acagttctcc gcaagaattg attggctcca
2761 attcttggag tggtgaatcc gttagcgagg tgccgccggc ttccattcag gtcgaggtgg
2821 cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg gcggcgccta
2881 caatccatgc caacccgttc catgtgctcg ccgaggcggc ataaatcgcc gtgacgatca
2941 gcggtccaat gatcgaagtt aggctggtaa gagccgcgag cgatccttga agctgtccct
3001 gatggtcgtc atctacctgc ctggacagca tggcctgcaa cgcgggcatc ccgatgccgc
3061 cggaagcgag aagaatcata atggggaagg ccatccagcc tcgcgtcgcg aacgccagca
3121 agacgtagcc cagcgcgtcg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac
3181 gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg
3241 caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc
3301 agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg
3361 cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
3421 gcatcggtcg acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt
3481 tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca
3541 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
3601 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
3661 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatccac aggacgggtg
3721 tggtcgccat gatcgcgtag tcgatagtgg ctccaagtag cgaagcgagc aggactgggc
3781 ggcggccaaa gcggtcggac agtgctccga gaacgggtgc gcatagaaat tgcatcaacg
3841 catatagcgc tagcagcacg ccatagtgac tggcgatgct gtcggaatgg acgatcacac
3901 tactcttctt ttgcctatta taacatgaaa acgctttaac agaaagaata ggaaggtatc
3961 cgactgctgg tattaaccct ctttctcaag ttatcatcgg caatgttgaa tggagtccat
4021 tcaaaacagc atagctctaa aacatggaac tcaacaagtc tcagtgtgct gaagttttgg
4081 gaccattcaa aacagcatag ctctaaaacg agacctttga gcttccgaga ctggtctcag
4141 ttttgggacc attcaaaaca gcatagctct aaaacctcgt agactatttt tgtctaaaaa
4201 atttcgtaat cgcactattt gtctcagcta gacttcagtc ttgaaaagcc cctgtattac
4261 tgcatttatt aagagtatta taccatattt ttagttatta agaaataatc ttcatctaaa
4321 atatacttca gtcacctcct agctgactca aatcaatgcg tgtttcataa agaccagtga
4381 tggattgatg gataagagtg gcatctaaaa cttcttttgt agacgtatat cgtttacgat
4441 caattgttgt atcaaaatat ttaaaagcag cgggagctcc aagattcgtc aacgtaaata
4501 aatgaataat attttctgct tgttcacgta ttggtttgtc tctatgtttg ttatatgcac
4561 taagaacttt atctaaattg gcatctgcta aaataacacg cttagaaaat tcactgattt
4621 gctcaataat ctcatctaaa taatgcttat gctgctccac aaacaattgt ttttgttcgt
4681 tatcttctgg actacccttc aacttttcat aatgactagc taaatataaa aaattcacat
4741 atttgcttgg cagagccagc tcatttcctt tttgtaattc tccggcacta gccagcatcc
4801 gtttacgacc gttttctaac tcaaaaagac tatatttagg tagtttaatg attaagtctt
4861 ttttaacttc cttatatcct ttagcttcta aaaagtcaat cggatttttt tcaaaggaac
4921 ttctttccat aattgtgatc cctagtaact ctttaacgga ttttaacttc ttcgatttcc
4981 ctttttccac cttagcaacc actaggactg aataagctac cgttggacta tcaaaaccac
5041 catatttttt tggatcccag tcttttttac gagcaataag cttgtccgaa tttctttttg
5101 gtaaaattga ctccttggag aatccgcctg tctgtacttc tgttttcttg acaatattga
5161 cttggggcat ggacaatact ttgcgcactg tggcaaaatc tcgcccttta tcccagacaa
5221 tttctccagt ttccccatta gtttcgatta gagggcgttt gcgaatctct ccatttgcaa
5281 gtgtaatttc tgttttgaag aagttcatga tattagagta aaagaaatat tttgcggttg
5341 ctttgcctat ttcttgctca gacttagcaa tcattttacg aacatcataa actttataat
5401 caccatagac aaactccgat tcaagttttg gatatttctt aatcaaagca gttccaacga
5461 cggcatttag atacgcatca tgggcatgat ggtaattgtt aatctcacgt actttataga
5521 attggaaatc ttttcggaag tcagaaacta atttagattt taaggtaatc actttaacct
5581 ctcgaataag tttatcattt tcatcgtatt tagtattcat gcgactatcc aaaatttgtg
5641 ccacatgctt agtgatttgg cgagtttcaa ccaattggcg tttgataaaa ccagctttat
5701 caagttcact caaacctcca cgttcagctt tcgttaaatt atcaaactta cgttgagtga
5761 ttaacttggc gtttagaagt tgtctccaat agtttttcat ctttttgact acttcttcac
5821 ttggaacgtt atccgattta ccacgatttt tatcagaacg cgttaagacc ttattgtcta
5881 ttgaatcgtc tttaaggaaa ctttgtggaa caatgtgatc gacatcataa tcacttaaac
5941 gattaatatc taattcttgg tccacataca tgtctcttcc attttggaga taatagagat
6001 agagcttttc attttgcaat tgagtatttt caacaggatg ctctttaaga atctgacttc
6061 ctaattcttt gataccttct tcgattcgtt tcatacgctc tcgcgaattt ttctggccct
6121 tttgagttgt ctgattttca cgtgccattt caataacgat attttctggc ttatgccgcc
6181 ccattacttt gaccaattca tcaacaactt ttacagtctg taaaatacct tttttaatag
6241 cagggctacc agctaaattt gcaatatgtt catgtaaact atcgccttgt ccagacactt
6301 gtgctttttg aatgtcttct ttaaatgtca aactatcatc atggatcagc tgcataaaat
6361 tgcgattggc aaaaccatct gatttcaaaa aatctaatat tgttttgcca gattgcttat
6421 ccctaatacc attaatcaat tttcgagaca aacgtcccca accagtataa cggcgacgtt
6481 taagctgttt catcacctta tcatcaaaga ggtgagcata tgttttaagt ctttcctcaa
6541 tcatctccct atcttcaaat aaggtcaatg ttaaaacaat atcctctaag atatcttcat
6601 tttcttcatt atccaaaaaa tctttatctt taataatttt tagcaaatca tggtaggtac
6661 ctaatgaagc attaaatcta tcttcaactc ctgaaatttc aacactatca aaacattcta
6721 tttttttgaa ataatcttct tttaattgct taacggttac ttttcgattt gttttgaaga
6781 gtaaatcaac aatggctttc ttctgttcac ctgaaagaaa tgctggtttt cgcattcctt
6841 cagtaacata tttgaccttt gtcaattcgt tataaaccgt aaaatactca taaagcaaac
6901 tatgttttgg tagtactttt tcatttggaa gatttttatc aaagtttgtc atgcgttcaa
6961 taaatgattg agctgaagca cctttatcga caacttcttc aaaattccat ggggtaattg
7021 tttcttcaga cttccgagtc atccatgcaa aacgactatt gccacgcgcc aatggaccaa
7081 cataataagg aattcgaaaa gtcaagattt tttcaatctt ctcacgattg tcttttaaaa
7141 atggataaaa gtcttcttgt cttctcaaaa tagcatgcag ctcacccaag tgaatttgat
7201 ggggaataga gccgttgtca aaggtccgtt gcttgcgcag caaatcttca cgatttagtt
7261 tcaccaataa ttcctcagta ccatccattt tttctaaaat tggtttgata aatttataaa
7321 attcttcttg gctagctccc ccatcaatat aacctgcata tccgtttttt gattgatcaa
7381 aaaagatttc tttatacttt tctggaagtt gttgtcgaac taaagctttt aaaagagtca
7441 agtcttgatg atgttcatcg tagcgtttaa tcattgaagc tgatagggga gccttagtta
7501 tttcagtatt tactcttagg atatctgaaa gtaaaatagc atctgataaa ttcttagctg
7561 ccaaaaacaa atcagcatat tgatctccaa tttgcgccaa taaattatct aaatcatcat
7621 cgtaagtatc ttttgaaagc tgtaatttag catcttctgc caaatcaaaa tttgatttaa
7681 aattaggggt caaacccaat gacaaagcaa tgagattccc aaataagcca tttttcttct
7741 caccggggag ctgagcaatg agattttcta atcgtcttga tttactcaat cgtgcagaaa
7801 gaatcgcttt agcatctact ccacttgcgt taatagggtt ttcttcaaat aattgattgt
7861 aggtttgtac caactggata aatagtttgt ccacatcact attatcagga tttaaatctc
7921 cctcaatcaa aaaatgacca cgaaacttaa tcatatgcgc taaggccaaa tagattaagc
7981 gcaaatccgc tttatcagta gaatctacca atttttttcg cagatgatag atagttggat
8041 atttctcatg ataagcaact tcatctacta tatttccaaa aataggatga cgttcatgct
8101 tcttgtcttc ttccaccaaa aaagactctt caagtcgatg aaagaaacta tcatctactt
8161 tcgccatctc atttgaaaaa atctcctgta gataacaaat acgattcttc cgacgtgtat
8221 accttctacg agctgtccgt ttgagacgag tcgcttccgc tgtctctcca ctgtcaaata
8281 aaagagcccc tataagattt tttttgatac tgtggcggtc tgtatttccc agaaccttga
8341 actttttaga cggaacctta tattcatcag tgatcaccgc ccatccgacg ctatttgtgc
8401 cgatatctaa gcctattgag tatttcttat ccatttttgc ctcctaaaat aaaaagttta
8461 aattaaatcc ataatgagtt tgatgatttc aataatagtt ttaatgacct ccgaaattag
8521 tttaatatgc tttaattttt ctttttcaaa atatctcttc aaaaaatatt acccaatact
8581 taataataaa tagattataa cacaaaattc ttttaaaaag tagtttattt tgttatcatt
8641 ctatagtatt aagtattgtt ttatggctga taaatttctt tgaatttctc cttgattatt
8701 tgttataaaa gttataaaat aatcttgttg gaaccattca aaacagcata gcaagttaaa
8761 ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt tttttgatac
8821 ttctattcta ctctgactgc aaaccaaaaa aacaagcgct ttcaaaacgc ttgttttatc
8881 atttttaggg aaattaatct cttaatcctt ttatcattct acatttaggc gctgccatct
8941 tgctaaacct actaagctcc acaggatgat ttcgtaatcc cgcaagaggc ccggcagtac
9001 cggcataacc aagcctatgc ctacagcatc cagggtgacg gtgccgagga tgacgatgag
9061 cgcattgtta gatttcatac acggtgcctg actgcgttag caatttaact gtgataaact
9121 accgcattaa agcttatcga tgataagctg tcaaacatga gaattacaac ttatatcgta
9181 tggggctgac ttcaggtgct acatttgaag agataaattg cactgaaatc tagaaatatt
9241 ttatctgatt aataagatga tcttcttgag atcgttttgg tctgcgcgta atctcttgct
9301 ctgaaaacga aaaaaccgcc ttgcag
//
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