• 规格：20μl质粒
• 原核抗性：氨苄青霉素Amp
• 克隆菌株：大肠杆菌Stbl3
• 质粒简介：          pXT7-Cas9是一个爪蟾基因编辑Cas9表达质粒。
• 质粒图谱1： png/1-1P11G3440b36.png

质粒序列： 点击查看序列

LOCUS       Exported                7354 bp ds-DNA     circular SYN 17-JAN-2018

DEFINITION  synthetic circular DNA

FEATURES             Location/Qualifiers

     source          1..7354

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     promoter        417..435

                     /label=SP6 promoter

                     /note="promoter for bacteriophage SP6 RNA polymerase"

     3'UTR           complement(541..666)

                     /label=Xenopus globin 3'-UTR

                     /note="3'-UTR of the major beta-globin gene of Xenopus 

                     laevis"

     CDS             complement(702..749)

                     /codon_start=1

                     /product="bipartite nuclear localization signal from 

                     nucleoplasmin"

                     /label=nucleoplasmin NLS

                     /translation="KRPAATKKAGQAKKKK"

     CDS             complement(750..4853)

                     /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             complement(4878..4898)

                     /codon_start=1

                     /product="nuclear localization signal of SV40 (simian virus

                     40) large T antigen"

                     /label=SV40 NLS

                     /translation="PKKKRKV"

     regulatory      4901..4910

                     /regulatory_class="other"

                     /note="vertebrate consensus sequence for strong initiation 

                     of translation (Kozak, 1987)"

     5'UTR           complement(4923..4965)

                     /label=Xenopus globin 5'-UTR

                     /note="translational enhancer from the 5'-UTR of the major 

                     beta-globin gene of Xenopus laevis"

     promoter        complement(4978..4996)

                     /label=T7 promoter

                     /note="promoter for bacteriophage T7 RNA polymerase"

     rep_origin      complement(5417..6005)

                     /direction=LEFT

                     /label=ori

                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 

                     replication"

     CDS             complement(6176..7036)

                     /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"

     promoter        complement(7037..7141)

                     /gene="bla"

                     /label=AmpR promoter

ORIGIN

        1 tagcgtatca gagtctcgtc cgtcggtcgt atgacggtga aacctctgac acatgcagct

       61 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg

      121 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat

      181 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata

      241 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg

      301 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg

      361 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattggattt

      421 aggtgacact atagaatacg aattaattcg agctcgcccg gggatcctct agagtcgacc

      481 tgcagggggg gggggggggg gttttttttt tttttttttt tttttttttt tttttagaat

      541 gtgaagaaac tttcttttta ttaggagcag atacgaatgg ctacattttg ggggacaaca

      601 ttttgtaaag tgtaagttgg tattatgtag cttagagact ccattcgggt gttcttgagg

      661 ctggtttagt ggtaaccaga tcctagtcag tcactagatt atttcttttt cttagcttga

      721 ccagctttct tagtagcagc aggacgcttg tctccaccga gctgagagag gtcgattctt

      781 gtttcataga gccccgtaat tgactgatga atcagtgtgg cgtccaggac ctcctttgta

      841 gaggtgtacc gctttctgtc tatggtggtg tcgaagtact tgaaggctgc aggcgcgccc

      901 aagttggtca gagtaaacaa gtggataatg ttttctgcct gctccctgat gggcttatcc

      961 ctgtgcttat tgtaagcaga aagcacctta tcgaggttag cgtcggcgag gatcactctt

     1021 ttggagaatt cgcttatttg ctcgatgatc tcatcaaggt agtgtttgtg ttgttccacg

     1081 aacagctgct tctgctcatt atcttcggga gaccctttga gcttttcata gtggctggcc

     1141 agatacaaga aattaacgta tttagagggc agtgccagct cgttaccttt ctgcagctcg

     1201 cccgcactag cgagcattcg tttccggccg ttttcaagct caaagagaga gtacttggga

     1261 agcttaatga tgaggtcttt tttgacctct ttatatcctt tcgcctcgag aaagtcgatg

     1321 gggttttttt cgaagcttga tcgctccatg attgtgatgc ccagcagttc cttgacgctt

     1381 ttgagttttt tagacttccc tttctccact ttggccacaa ccagtacact gtaagcgact

     1441 gtaggagaat cgaatccgcc gtatttcttg gggtcccaat cttttttgcg tgcgatcagc

     1501 ttgtcgctgt tccttttcgg gaggatactt tccttggaga agcctccggt ctgtacttcg

     1561 gtctttttaa cgatgttcac ctgcggcatg gacaggacct tccggactgt cgcgaaatcc

     1621 ctacccttgt cccacacgat ttctcctgtt tctccgtttg tttcgataag tggtcgcttc

     1681 cgaatctctc cattggccag tgtaatctcg gtcttgaaaa aattcataat attgctgtaa

     1741 aagaagtact tagcggtggc cttgcctatt tcctgctcag actttgcgat cattttccta

     1801 acatcgtaca ctttatagtc tccgtaaaca aattcagatt caagcttggg atattttttg

     1861 ataagtgcag tgcctaccac tgcattcagg taggcatcat gcgcatggtg gtaattgttg

     1921 atctctctca ccttataaaa ctgaaagtcc tttctgaaat ctgagaccag cttagacttc

     1981 agagtaataa ctttcacctc tcgaatcagt ttgtcatttt catcgtactt ggtgttcatg

     2041 cgtgaatcga gaatttgggc cacgtgcttg gtgatctggc gtgtctcaac aagctgcctt

     2101 ttgatgaagc ctgctttatc caactcagac aggccacctc gttcagcctt agtcagatta

     2161 tcgaacttcc gttgtgtgat cagtttggcg ttcagcagct gccgccaata atttttcatt

     2221 ttcttgacaa cttcttctga ggggacgtta tcactcttcc ctctattttt atcggatctt

     2281 gtcaacactt tattatcaat agaatcatct ttgagaaaag actggggcac gatatgatcc

     2341 acgtcgtagt cggagagccg attgatgtcc agttcctgat ccacgtacat gtccctgccg

     2401 ttctgcaggt agtacaggta gagcttctca ttctgaagct gggtgttttc aactgggtgt

     2461 tccttaagga tttgggaccc cagttctttt ataccctctt caatcctctt catcctttcc

     2521 ctactgttct tctgtccctt ctgggtagtt tggttctctc gggccatctc gataacgata

     2581 ttctcgggct tatgccttcc cattactttg acgagttcat ccacgacctt aacggtctgc

     2641 agtattccct ttttgatagc tgggctacct gcaagattag cgatgtgctc gtgaagactg

     2701 tccccctggc cagaaacttg tgctttctgg atgtcctcct taaaggtgag agagtcatca

     2761 tggatcaact gcatgaagtt ccggttggca aatccatcgg acttaagaaa atccaggatt

     2821 gtctttccac tctgcttgtc tcggatccca ttgatcagtt ttcttgacag ccgcccccat

     2881 cctgtatatc ggcgcctctt gagctgtttc atgactttgt cgtcgaagag atgagcgtaa

     2941 gttttcaagc gttcttcaat catctcccta tcttcaaaca acgtaagggt gaggacaatg

     3001 tcctcaagaa tgtcctcgtt ctcctcattg tccaggaagt ccttgtcttt aatgattttc

     3061 aggagatcgt gatacgttcc cagggatgcg ttgaagcgat cctccactcc gctgatttca

     3121 acagagtcga aacattcaat ctttttgaaa tagtcttctt tgagctgttt cacggtaact

     3181 ttccggttcg tcttgaagag gaggtccacg atagctttct tctgctctcc agacaggaat

     3241 gctggctttc tcatcccttc tgtgacgtat ttgaccttgg tgagctcgtt ataaactgtg

     3301 aagtactcgt acagcagaga gtgtttagga agcacctttt cgttaggcag atttttatca

     3361 aagttagtca tcctttcgat gaaggactgg gcagaggccc ccttatccac gacttcctcg

     3421 aagttccagg gagtgatggt ctcttctgat ttgcgagtca tccacgcgaa tctggaattt

     3481 ccccgggcga gggggcctac atagtagggt atccgaaatg tgaggatttt ctcaatcttt

     3541 tccctgttat ctttcaaaaa ggggtagaaa tcctcttgcc gcctgagtat agcgtgcagt

     3601 tcgcccaggt gaatctggtg ggggatgctt ccattgtcga aagtgcgctg tttgcgcaac

     3661 agatcttctc tgttaagctt taccagcagc tcctcggtgc cgtccatttt ttccaagatg