• 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：          MSP680是一个哺乳CAS9表达质粒。
• 质粒图谱1： png/1-1G204153922617.png

质粒序列： 点击查看序列

LOCUS       Exported                7614 bp ds-DNA     circular SYN 04-DEC-2017

DEFINITION  synthetic circular DNA

FEATURES             Location/Qualifiers

     source          1..7614

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     promoter        132..335

                     /label=CMV promoter

                     /note="human cytomegalovirus (CMV) immediate early 

                     promoter"

     promoter        377..395

                     /label=T7 promoter

                     /note="promoter for bacteriophage T7 RNA polymerase"

     regulatory      403..412

                     /regulatory_class="other"

                     /note="vertebrate consensus sequence for strong initiation 

                     of translation (Kozak, 1987)"

     CDS             409..4512

                     /codon_start=1

                     /product="Streptococcus pyogenes Cas9 endonuclease with the

                     D1135V/R1335Q/T1337R mutations"

                     /label=Cas9 VQR

                     /note="has an altered recognition specificity for the 

                     protospacer adjacent motif (PAM) (Kleinstiver et al., 

                     2015)"

                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK

                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES

                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK

                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL

                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ

                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR

                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL

                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG

                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY

                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD

                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER

                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM

                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR

                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY

                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP

                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH

                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL

                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT

                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK

                     ESILPKRNSDKLIARKKDWDPKKYGGFESPTVAYSVLVVAKVEKGKSKKLKSVKELLGI

                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE

                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD

                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKQYRSTKEVL

                     DATLIHQSITGLYETRIDLSQLGGD"

     CDS             4519..4539

                     /codon_start=1

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

                     40) large T antigen"

                     /label=SV40 NLS

                     /translation="PKKKRKV"

     CDS             4546..4611

                     /codon_start=1

                     /product="three tandem FLAG(R) epitope tags, followed by an

                     enterokinase cleavage site"

                     /label=3xFLAG

                     /translation="DYKDHDGDYKDHDIDYKDDDDK"

     polyA_signal    4675..4899

                     /label=bGH poly(A) signal

                     /note="bovine growth hormone polyadenylation signal"

     rep_origin      complement(5372..5960)

                     /direction=LEFT

                     /label=ori

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

                     replication"

     CDS             complement(6131..6991)

                     /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(6992..7096)

                     /gene="bla"

                     /label=AmpR promoter

     enhancer        7366..131

                     /label=CMV enhancer

                     /note="human cytomegalovirus immediate early enhancer"

ORIGIN

        1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg

       61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg

      121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact

      181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa

      241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta

      301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct

      361 agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat ggataaaaag

      421 tattctattg gtttagacat cggcactaat tccgttggat gggctgtcat aaccgatgaa

      481 tacaaagtac cttcaaagaa atttaaggtg ttggggaaca cagaccgtca ttcgattaaa

      541 aagaatctta tcggtgccct cctattcgat agtggcgaaa cggcagaggc gactcgcctg

      601 aaacgaaccg ctcggagaag gtatacacgt cgcaagaacc gaatatgtta cttacaagaa

      661 atttttagca atgagatggc caaagttgac gattctttct ttcaccgttt ggaagagtcc

      721 ttccttgtcg aagaggacaa gaaacatgaa cggcacccca tctttggaaa catagtagat

      781 gaggtggcat atcatgaaaa gtacccaacg atttatcacc tcagaaaaaa gctagttgac

      841 tcaactgata aagcggacct gaggttaatc tacttggctc ttgcccatat gataaagttc

      901 cgtgggcact ttctcattga gggtgatcta aatccggaca actcggatgt cgacaaactg

      961 ttcatccagt tagtacaaac ctataatcag ttgtttgaag agaaccctat aaatgcaagt

     1021 ggcgtggatg cgaaggctat tcttagcgcc cgcctctcta aatcccgacg gctagaaaac

     1081 ctgatcgcac aattacccgg agagaagaaa aatgggttgt tcggtaacct tatagcgctc

     1141 tcactaggcc tgacaccaaa ttttaagtcg aacttcgact tagctgaaga tgccaaattg

     1201 cagcttagta aggacacgta cgatgacgat ctcgacaatc tactggcaca aattggagat

     1261 cagtatgcgg acttattttt ggctgccaaa aaccttagcg atgcaatcct cctatctgac

     1321 atactgagag ttaatactga gattaccaag gcgccgttat ccgcttcaat gatcaaaagg

     1381 tacgatgaac atcaccaaga cttgacactt ctcaaggccc tagtccgtca gcaactgcct

     1441 gagaaatata aggaaatatt ctttgatcag tcgaaaaacg ggtacgcagg ttatattgac

     1501 ggcggagcga gtcaagagga attctacaag tttatcaaac ccatattaga gaagatggat

     1561 gggacggaag agttgcttgt aaaactcaat cgcgaagatc tactgcgaaa gcagcggact

     1621 ttcgacaacg gtagcattcc acatcaaatc cacttaggcg aattgcatgc tatacttaga

     1681 aggcaggagg atttttatcc gttcctcaaa gacaatcgtg aaaagattga gaaaatccta

     1741 acctttcgca taccttacta tgtgggaccc ctggcccgag ggaactctcg gttcgcatgg

     1801 atgacaagaa agtccgaaga aacgattact ccatggaatt ttgaggaagt tgtcgataaa

     1861 ggtgcgtcag ctcaatcgtt catcgagagg atgaccaact ttgacaagaa tttaccgaac

     1921 gaaaaagtat tgcctaagca cagtttactt tacgagtatt tcacagtgta caatgaactc

     1981 acgaaagtta agtatgtcac tgagggcatg cgtaaacccg cctttctaag cggagaacag

     2041 aagaaagcaa tagtagatct gttattcaag accaaccgca aagtgacagt taagcaattg

     2101 aaagaggact actttaagaa aattgaatgc ttcgattctg tcgagatctc cggggtagaa

     2161 gatcgattta atgcgtcact tggtacgtat catgacctcc taaagataat taaagataag

     2221 gacttcctgg ataacgaaga gaatgaagat atcttagaag atatagtgtt gactcttacc

     2281 ctctttgaag atcgggaaat gattgaggaa agactaaaaa catacgctca cctgttcgac

     2341 gataaggtta tgaaacagtt aaagaggcgt cgctatacgg gctggggacg attgtcgcgg

     2401 aaacttatca acgggataag agacaagcaa agtggtaaaa ctattctcga ttttctaaag

     2461 agcgacggct tcgccaatag gaactttatg cagctgatcc atgatgactc tttaaccttc

     2521 aaagaggata tacaaaaggc acaggtttcc ggacaagggg actcattgca cgaacatatt

     2581 gcgaatcttg ctggttcgcc agccatcaaa aagggcatac tccagacagt caaagtagtg

     2641 gatgagctag ttaaggtcat gggacgtcac aaaccggaaa acattgtaat cgagatggca

     2701 cgcgaaaatc aaacgactca gaaggggcaa aaaaacagtc gagagcggat gaagagaata

     2761 gaagagggta ttaaagaact gggcagccag atcttaaagg agcatcctgt ggaaaatacc

     2821 caattgcaga acgagaaact ttacctctat tacctacaaa atggaaggga catgtatgtt

     2881 gatcaggaac tggacataaa ccgtttatct gattacgacg tcgatcacat tgtaccccaa

     2941 tcctttttga aggacgattc aatcgacaat aaagtgctta cacgctcgga taagaaccga

     3001 gggaaaagtg acaatgttcc aagcgaggaa gtcgtaaaga aaatgaagaa ctattggcgg

     3061 cagctcctaa atgcgaaact gataacgcaa agaaagttcg ataacttaac taaagctgag

     3121 aggggtggct tgtctgaact tgacaaggcc ggatttatta aacgtcagct cgtggaaacc

     3181 cgccaaatca caaagcatgt tgcacagata ctagattccc gaatgaatac gaaatacgac

     3241 gagaacgata agctgattcg ggaagtcaaa gtaatcactt taaagtcaaa attggtgtcg

     3301 gacttcagaa aggattttca attctataaa gttagggaga taaataacta ccaccatgcg

     3361 cacgacgctt atcttaatgc cgtcgtaggg accgcactca ttaagaaata cccgaagcta

     3421 gaaagtgagt ttgtgtatgg tgattacaaa gtttatgacg tccgtaagat gatcgcgaaa

     3481 agcgaacagg agataggcaa ggctacagcc aaatacttct tttattctaa cattatgaat

     3541 ttctttaaga cggaaatcac tctggcaaac ggagagatac gcaaacgacc tttaattgaa

     3601 accaatgggg agacaggtga aatcgtatgg gataagggcc gggacttcgc gacggtgaga