• 规格：10μl质粒
• 质粒图谱1： png/1-1F92Q1394UY.png

质粒序列： 点击查看序列

LOCUS       Exported                8783 bp ds-DNA     circular SYN 28-SEP-2017

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    pRH2502

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 8783)

  AUTHORS   .

  TITLE     Direct Submission

  JOURNAL   Exported Sep 28, 2017 from MiaoLingPlasmid

            http://www.fenghbio.cn

FEATURES             Location/Qualifiers

     source          1..8783

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     CDS             1..4104

                     /codon_start=1

                     /product="catalytically dead mutant of the Cas9 

                     endonuclease from the Streptococcus pyogenes Type II 

                     CRISPR/Cas system"

                     /label=dCas9

                     /note="RNA-guided DNA-binding protein that lacks 

                     endonuclease activity due to the D10A mutation in the RuvC 

                     catalytic domain and the H840A mutation in the HNH 

                     catalytic domain"

                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK

                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES

                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK

                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL

                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ

                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR

                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL

                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG

                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY

                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD

                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER

                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM

                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR

                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY

                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP

                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH

                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL

                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT

                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK

                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI

                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE

                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD

                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL

                     DATLIHQSITGLYETRIDLSQLGGD"

     CDS             4117..4137

                     /codon_start=1

                     /product="nuclear localization signal of SV40 large T 

                     antigen"

                     /label=SV40 NLS

                     /translation="PKKKRKV"

     polyA_signal    4158..4292

                     /label=SV40 poly(A) signal

                     /note="SV40 polyadenylation signal"

     rep_origin      complement(4488..5076)

                     /direction=LEFT

                     /label=ori

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

                     replication"

     CDS             complement(5407..6222)

                     /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

                     KPHAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK

                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA

                     SDFDDERNGWPVEQVWKEMHNLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI

                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"

     CDS             complement(6522..7520)

                     /codon_start=1

                     /label=Int

                     /translation="MDAEAWLAGEKRLIEMETWTPPQDRAKKAAASAITLEEYTRKWLV

                     ERDLADGTRDLYSGHAERRIYPVLGEVAVTEMTPALVRAWWAGMGRKHPTARRHAYNVL

                     RAVMNTAVEDKLIAENPCRIEQKAADERDVEALTPEELDIVAAEIFVHYRIAAYILAWT

                     SLRFGELIELRRKDIVDDGMTMKLRVRRGASRVGNKIVVGNAKTVRSKRPVTVPPHVAE

                     MIRAHMKDRTKMNKGPEAFLVTTTQGNRLSKSAFTKSLKRGYAKIGRPELRIHDLRAVG

                     ATFAAQAGATTKELMARLGHTTPRMAMKYQMASEARDEAIAEAMSKLAKTS"

     terminator      8170..8216

                     /label=rrnB T1 terminator

                     /note="transcription terminator T1 from the E. coli rrnB 

                     gene"

     misc_feature    8449..8625

                     /label=uvtetO promoter

     protein_bind    8626..8643

                     /gene="tetO"

                     /label=tet operator

                     /bound_moiety="tetracycline repressor TetR"

                     /note="bacterial operator O2 for the tetR and tetA genes"

ORIGIN

        1 atggacaaga agtacagcat cggcctggcg atcggcacga actcggtggg ctgggcggtc

       61 atcacggacg agtacaaggt cccctccaag aagttcaagg tcctgggcaa caccgaccgg

      121 cacagcatca agaagaacct gatcggcgcc ctgctgttcg actccggcga gaccgccgag

      181 gcgacgcgcc tgaagcggac cgcgcggcgc cgatacacgc ggcgcaagaa ccgcatctgc

      241 tacctgcagg aaatcttctc gaacgagatg gccaaggtgg acgacagctt cttccaccgg

      301 ctggaggagt ccttcctggt ggaggaggac aagaagcacg agcgccaccc gatcttcggc

      361 aacatcgtgg acgaggtggc ctaccacgag aagtacccca cgatctacca cctgcgcaag

      421 aagctggtgg acagcaccga caaggcggac ctgcggctga tctacctggc cctggcgcac

      481 atgatcaagt tccgcggcca cttcctgatc gagggcgacc tgaacccgga caactcggac

      541 gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggagaacccc

      601 atcaacgcct ccggcgtgga cgccaaggcg atcctgtcgg cgcgcctgtc caagtcgcgg

      661 cgcctggaga acctgatcgc ccagctgccg ggcgagaaga agaacggcct gttcggcaac

      721 ctgatcgcgc tgagcctggg cctgacgccc aacttcaagt ccaacttcga cctggccgag

      781 gacgcgaagc tgcagctgtc gaaggacacc tacgacgacg acctggacaa cctgctggcc

      841 cagatcggcg accagtacgc ggacctgttc ctggccgcca agaacctgag cgacgccatc

      901 ctgctgtccg acatcctgcg ggtcaacacc gagatcacga aggccccgct gagcgcgtcc

      961 atgatcaagc ggtacgacga gcaccaccag gacctgaccc tgctgaaggc gctggtgcgc

     1021 cagcagctgc ccgagaagta caaggaaatc ttcttcgacc agtcgaagaa cggctacgcc

     1081 ggctacatcg acggcggcgc gagccaggag gagttctaca agttcatcaa gccgatcctg

     1141 gagaagatgg acggcacgga ggagctgctg gtcaagctga accgcgagga cctgctgcgc

     1201 aagcagcgga ccttcgacaa cggctcgatc ccgcaccaga tccacctggg cgagctgcac

     1261 gccatcctgc ggcgccagga ggacttctac cccttcctga aggacaaccg cgagaagatc

     1321 gagaagatcc tgaccttccg catcccgtac tacgtgggcc ccctggcccg cggcaactcg

     1381 cggttcgcgt ggatgacccg gaagagcgag gagaccatca cgccgtggaa cttcgaggag

     1441 gtcgtggaca agggcgcctc ggcgcagagc ttcatcgagc gcatgaccaa cttcgacaag

     1501 aacctgccga acgagaaggt cctgcccaag cacagcctgc tgtacgagta cttcaccgtg

     1561 tacaacgagc tgacgaaggt caagtacgtg accgagggca tgcggaagcc ggccttcctg

     1621 tcgggcgagc agaagaaggc gatcgtggac ctgctgttca agaccaaccg caaggtcacg

     1681 gtgaagcagc tgaaggagga ctacttcaag aagatcgagt gtttcgactc ggtcgagatc

     1741 agcggcgtgg aggaccgctt caacgccagc ctgggcacct accacgacct gctgaagatc

     1801 atcaaggaca aggacttcct ggacaacgag gagaacgagg acatcctgga ggacatcgtc

     1861 ctgaccctga cgctgttcga ggaccgcgag atgatcgagg agcggctgaa gacgtacgcc

     1921 cacctgttcg acgacaaggt gatgaagcag ctgaagcggc gccgatacac cggctggggc

     1981 cgcctgtcgc ggaagctgat caacggcatc cgggacaagc agtccggcaa gaccatcctg

     2041 gacttcctga agtcggacgg cttcgcgaac cgcaacttca tgcagctgat ccacgacgac

     2101 agcctgacct tcaaggagga catccagaag gcccaagtga gcggccaggg cgactccctg

     2161 cacgagcaca tcgccaacct ggcgggctcg ccggcgatca agaagggcat cctgcagacc

     2221 gtcaaggtcg tggacgagct ggtcaaggtg atgggccgcc acaagcccga gaacatcgtg

     2281 atcgagatgg cccgggagaa ccagaccacg cagaagggcc agaagaacag ccgcgagcgg

     2341 atgaagcgca tcgaggaggg catcaaggag ctgggctcgc agatcctgaa ggagcacccg

     2401 gtcgagaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaacggccgc

     2461 gacatgtacg tggaccagga gctggacatc aaccggctga gcgactacga cgtggacgcc

     2521 atcgtgccgc agtcgttcct gaaggacgac agcatcgaca acaaggtcct gacccgctcg

     2581 gacaagaacc ggggcaagtc ggacaacgtg cccagcgagg aggtcgtgaa gaagatgaag

     2641 aactactggc gccagctgct gaacgccaag ctgatcacgc agcgcaagtt cgacaacctg

     2701 accaaggccg agcggggcgg cctgagcgag ctggacaagg cgggcttcat caagcgccag

     2761 ctggtcgaga cccggcagat cacgaagcac gtggcgcaga tcctggactc gcggatgaac

     2821 acgaagtacg acgagaacga caagctgatc cgcgaggtca aggtgatcac cctgaagtcc

     2881 aagctggtct cggacttccg caaggacttc cagttctaca aggtccggga gatcaacaac

     2941 taccaccacg cccacgacgc gtacctgaac gccgtcgtgg gcaccgcgct gatcaagaag

     3001 tacccgaagc tggagtcgga gttcgtctac ggcgactaca aggtctacga cgtgcgcaag

     3061 atgatcgcca agagcgagca ggagatcggc aaggccaccg cgaagtactt cttctactcc

     3121 aacatcatga acttcttcaa gaccgagatc acgctggcca acggcgagat ccgcaagcgc

     3181 ccgctgatcg agaccaacgg cgagacgggc gagatcgtct gggacaaggg ccgcgacttc

     3241 gcgaccgtcc ggaaggtgct gagcatgccg caggtcaaca tcgtgaagaa gaccgaggtg<