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載體描述

植物表達(dá)載體。

載體序列

LOCUS       Exported                8958 bp ds-DNA     circular SYN 22-MAR-2019

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    .

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 8958)

  AUTHORS   BT-Lab

  TITLE     Direct Submission

  JOURNAL   Exported Friday, Mar 22, 2019 from SnapGene Viewer 4.3.4

FEATURES             Location/Qualifiers

     source          1..8958

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     misc_feature    1..56

                     /label=MCS

                     /note="Multiple cloning site"

     primer_bind     complement(60..76)

                     /label=M13 fwd

                     /note="common sequencing primer, one of multiple similar

                     variants"

     misc_feature    279..303

                     /label=RB T-DNA repeat

                     /note="right border repeat from nopaline C58 T-DNA"

     CDS             1603..2232

                     /codon_start=1

                     /product="stability protein from the Pseudomonas plasmid

                     pVS1 (Heeb et al., 2000)"

                     /label=pVS1 StaA

                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR

                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP

                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI

                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"

     CDS             2666..3733

                     /codon_start=1

                     /product="replication protein from the Pseudomonas plasmid

                     pVS1 (Heeb et al., 2000)"

                     /label=pVS1 RepA

                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA

                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR

                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR

                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL

                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA

                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV

                     MRYRNLIEGEASAGS"

     rep_origin      3799..3993

                     /label=pVS1 oriV

                     /note="origin of replication for the Pseudomonas plasmid

                     pVS1 (Heeb et al., 2000)"

     misc_feature    4337..4477

                     /label=bom

                     /note="basis of mobility region from pBR322"

     rep_origin      complement(4663..5251)

                     /direction=LEFT

                     /label=ori

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

                     replication"

     CDS             complement(5338..6132)

                     /codon_start=1

                     /gene="aphA-3"

                     /product="aminoglycoside phosphotransferase"

                     /label=KanR

                     /note="confers resistance to kanamycin"

                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM

                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED

                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT

                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA

                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"

     misc_feature    6557..6581

                     /label=LB T-DNA repeat

                     /note="left border repeat from nopaline C58 T-DNA"

     polyA_signal    6659..6833

                     /label=CaMV poly(A) signal

                     /note="cauliflower mosaic virus polyadenylation signal"

     CDS             complement(6873..7898)

                     /codon_start=1

                     /gene="aph(4)-Ia"

                     /product="aminoglycoside phosphotransferase from E. coli"

                     /label=HygR

                     /note="confers resistance to hygromycin"

                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG

                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP

                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ

                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF

                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG

                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK

                     K"

     promoter        complement(7966..8643)

                     /label=CaMV 35S promoter (enhanced)

                     /note="cauliflower mosaic virus 35S promoter with a

                     duplicated enhancer region"

     protein_bind    8834..8855

                     /label=CAP binding site

                     /bound_moiety="E. coli catabolite activator protein"

                     /note="CAP binding activates transcription in the presence

                     of cAMP."

     promoter        8870..8900

                     /label=lac promoter

                     /note="promoter for the E. coli lac operon"

     protein_bind    8908..8924

                     /label=lac operator

                     /bound_moiety="lac repressor encoded by lacI"

                     /note="The lac repressor binds to the lac operator to

                     inhibit transcription in E. coli. This inhibition can be

                     relieved by adding lactose or

                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."

     primer_bind     8932..8948

                     /label=M13 rev

                     /note="common sequencing primer, one of multiple similar

                     variants"

ORIGIN

        1 aattcgagct cggtacccgg ggatcctcta gagtcgacct gcaggcatgc aagcttggca

       61 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc

      121 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc

      181 ccttcccaac agttgcgcag cctgaatggc gaatgctaga gcagcttgag cttggatcag

      241 attgtcgttt cccgccttca gtttaaacta tcagtgtttg acaggatata ttggcgggta

      301 aacctaagag aaaagagcgt ttattagaat aacggatatt taaaagggcg tgaaaaggtt

      361 tatccgttcg tccatttgta tgtgcatgcc aaccacaggg ttcccctcgg gatcaaagta

      421 ctttgatcca acccctccgc tgctatagtg cagtcggctt ctgacgttca gtgcagccgt

      481 cttctgaaaa cgacatgtcg cacaagtcct aagttacgcg acaggctgcc gccctgccct

      541 tttcctggcg ttttcttgtc gcgtgtttta gtcgcataaa gtagaatact tgcgactaga

      601 accggagaca ttacgccatg aacaagagcg ccgccgctgg cctgctgggc tatgcccgcg

      661 tcagcaccga cgaccaggac ttgaccaacc aacgggccga actgcacgcg gccggctgca

      721 ccaagctgtt ttccgagaag atcaccggca ccaggcgcga ccgcccggag ctggccagga

      781 tgcttgacca cctacgccct ggcgacgttg tgacagtgac caggctagac cgcctggccc

      841 gcagcacccg cgacctactg gacattgccg agcgcatcca ggaggccggc gcgggcctgc

      901 gtagcctggc agagccgtgg gccgacacca ccacgccggc cggccgcatg gtgttgaccg

      961 tgttcgccgg cattgccgag ttcgagcgtt ccctaatcat cgaccgcacc cggagcgggc

     1021 gcgaggccgc caaggcccga ggcgtgaagt ttggcccccg ccctaccctc accccggcac

     1081 agatcgcgca cgcccgcgag ctgatcgacc aggaaggccg caccgtgaaa gaggcggctg

     1141 cactgcttgg cgtgcatcgc tcgaccctgt accgcgcact tgagcgcagc gaggaagtga

     1201 cgcccaccga ggccaggcgg cgcggtgcct tccgtgagga cgcattgacc gaggccgacg

     1261 ccctggcggc cgccgagaat gaacgccaag aggaacaagc atgaaaccgc accaggacgg

     1321 ccaggacgaa ccgtttttca ttaccgaaga gatcgaggcg gagatgatcg cggccgggta

     1381 cgtgttcgag ccgcccgcgc acgtctcaac cgtgcggctg catgaaatcc tggccggttt

     1441 gtctgatgcc aagctggcgg cctggccggc cagcttggcc gctgaagaaa ccgagcgccg

     1501 ccgtctaaaa aggtgatgtg tatttgagta aaacagcttg cgtcatgcgg tcgctgcgta

     1561 tatgatgcga tgagtaaata aacaaatacg caaggggaac gcatgaaggt tatcgctgta

     1621 cttaaccaga aaggcgggtc aggcaagacg accatcgcaa cccatctagc ccgcgccctg

     1681 caactcgccg gggccgatgt tctgttagtc gattccgatc cccagggcag tgcccgcgat

     1741 tgggcggccg tgcgggaaga tcaaccgcta accgttgtcg gcatcgaccg cccgacgatt

     1801 gaccgcgacg tgaaggccat cggccggcgc gacttcgtag tgatcgacgg agcgccccag

     1861 gcggcggact tggctgtgtc cgcgatcaag gcagccgact tcgtgctgat tccggtgcag

     1921 ccaagccctt acgacatatg ggccaccgcc gacctggtgg agctggttaa gcagcgcatt

     1981 gaggtcacgg atggaaggct acaagcggcc tttgtcgtgt cgcgggcgat caaaggcacg

     2041 cgcatcggcg gtgaggttgc cgaggcgctg gccgggtacg agctgcccat tcttgagtcc

     2101 cgtatcacgc agcgcgtgag ctacccaggc actgccgccg ccggcacaac cgttcttgaa

     2161 tcagaacccg agggcgacgc tgcccgcgag gtccaggcgc tggccgctga aattaaatca

     2221 aaactcattt gagttaatga ggtaaagaga aaatgagcaa aagcacaaac acgctaagtg

     2281 ccggccgtcc gagcgcacgc agcagcaagg ctgcaacgtt ggccagcctg gcagacacgc

     2341 cagccatgaa gcgggtcaac tttcagttgc cggcggagga tcacaccaag ctgaagatgt

     2401 acgcggtacg ccaaggcaag accattaccg agctgctatc tgaatacatc gcgcagctac

     2461 cagagtaaat gagcaaatga ataaatgagt agatgaattt tagcggctaa aggaggcggc

     2521 atggaaaatc aagaacaacc aggcaccgac gccgtggaat gccccatgtg tggaggaacg

     2581 ggcggttggc caggcgtaag cggctgggtt gtctgccggc cctgcaatgg cactggaacc

     2641 cccaagcccg aggaatcggc gtgacggtcg caaaccatcc ggcccggtac aaatcggcgc

     2701 ggcgctgggt gatgacctgg tggagaagtt gaaggccgcg caggccgccc agcggcaacg

     2761 catcgaggca gaagcacgcc ccggtgaatc gtggcaagcg gccgctgatc gaatccgcaa

     2821 agaatcccgg caaccgccgg cagccggtgc gccgtcgatt aggaagccgc ccaagggcga

     2881 cgagcaacca gattttttcg ttccgatgct ctatgacgtg ggcacccgcg atagtcgcag

     2941 catcatggac gtggccgttt tccgtctgtc gaagcgtgac cgacgagctg gcgaggtgat

     3001 ccgctacgag cttccagacg ggcacgtaga ggtttccgca gggccggccg gcatggccag

     3061 tgtgtgggat tacgacctgg tactgatggc ggtttcccat ctaaccgaat ccatgaaccg

     3121 ataccgggaa gggaagggag acaagcccgg ccgcgtgttc cgtccacacg ttgcggacgt

     3181 actcaagttc tgccggcgag ccgatggcgg aaagcagaaa gacgacctgg tagaaacctg

     3241 cattcggtta aacaccacgc acgttgccat gcagcgtacg aagaaggcca agaacggccg

     3301 cctggtgacg gtatccgagg gtgaagcctt gattagccgc tacaagatcg taaagagcga

     3361 aaccgggcgg ccggagtaca tcgagatcga gctagctgat tggatgtacc gcgagatcac

     3421 agaaggcaag aacccggacg tgctgacggt tcaccccgat tactttttga tcgatcccgg

     3481 catcggccgt tttctctacc gcctggcacg ccgcgccgca ggcaaggcag aagccagatg

     3541 gttgttcaag acgatctacg aacgcagtgg cagcgccgga gagttcaaga agttctgttt

     3601 caccgtgcgc aagctgatcg ggtcaaatga cctgccggag tacgatttga aggaggaggc

     3661 ggggcaggct ggcccgatcc tagtcatgcg ctaccgcaac ctgatcgagg gcgaagcatc

     3721 cgccggttcc taatgtacgg agcagatgct agggcaaatt gccctagcag gggaaaaagg

     3781 tcgaaaaggt ctctttcctg tggatagcac gtacattggg aacccaaagc cgtacattgg

     3841 gaaccggaac ccgtacattg ggaacccaaa gccgtacatt gggaaccggt cacacatgta

     3901 agtgactgat ataaaagaga aaaaaggcga tttttccgcc taaaactctt taaaacttat

     3961 taaaactctt aaaacccgcc tggcctgtgc ataactgtct ggccagcgca cagccgaaga

     4021 gctgcaaaaa gcgcctaccc ttcggtcgct gcgctcccta cgccccgccg cttcgcgtcg

     4081 gcctatcgcg gccgctggcc gctcaaaaat ggctggccta cggccaggca atctaccagg

     4141 gcgcggacaa gccgcgccgt cgccactcga ccgccggcgc ccacatcaag gcaccctgcc

     4201 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca

     4261 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg

     4321 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg

     4381 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat

     4441 accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac

     4501 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt

     4561 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca

     4621 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc

     4681 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact

     4741 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct

     4801 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag

     4861 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca

     4921 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa

     4981 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc

     5041 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag

     5101 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg

     5161 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca

     5221 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc

     5281 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgcatt ctaggtacta

     5341 aaacaattca tccagtaaaa tataatattt tattttctcc caatcaggct tgatccccag

     5401 taagtcaaaa aatagctcga catactgttc ttccccgata tcctccctga tcgaccggac

     5461 gcagaaggca atgtcatacc acttgtccgc cctgccgctt ctcccaagat caataaagcc

     5521 acttactttg ccatctttca caaagatgtt gctgtctccc aggtcgccgt gggaaaagac

     5581 aagttcctct tcgggctttt ccgtctttaa aaaatcatac agctcgcgcg gatctttaaa

     5641 tggagtgtct tcttcccagt tttcgcaatc cacatcggcc agatcgttat tcagtaagta

     5701 atccaattcg gctaagcggc tgtctaagct attcgtatag ggacaatccg atatgtcgat

     5761 ggagtgaaag agcctgatgc actccgcata cagctcgata atcttttcag ggctttgttc

     5821 atcttcatac tcttccgagc aaaggacgcc atcggcctca ctcatgagca gattgctcca

     5881 gccatcatgc cgttcaaagt gcaggacctt tggaacaggc agctttcctt ccagccatag

     5941 catcatgtcc ttttcccgtt ccacatcata ggtggtccct ttataccggc tgtccgtcat

     6001 ttttaaatat aggttttcat tttctcccac cagcttatat accttagcag gagacattcc

     6061 ttccgtatct tttacgcagc ggtatttttc gatcagtttt ttcaattccg gtgatattct

     6121 cattttagcc atttattatt tccttcctct tttctacagt atttaaagat accccaagaa

     6181 gctaattata acaagacgaa ctccaattca ctgttccttg cattctaaaa ccttaaatac

     6241 cagaaaacag ctttttcaaa gttgttttca aagttggcgt ataacatagt atcgacggag

     6301 ccgattttga aaccgcggtg atcacaggca gcaacgctct gtcatcgtta caatcaacat

     6361 gctaccctcc gcgagatcat ccgtgtttca aacccggcag cttagttgcc gttcttccga

     6421 atagcatcgg taacatgagc aaagtctgcc gccttacaac ggctctcccg ctgacgccgt

     6481 cccggactga tgggctgcct gtatcgagtg gtgattttgt gccgagctgc cggtcgggga

     6541 gctgttggct ggctggtggc aggatatatt gtggtgtaaa caaattgacg cttagacaac

     6601 ttaataacac attgcggacg tttttaatgt actgaattaa cgccgaatta attcggggga

     6661 tctggatttt agtactggat tttggtttta ggaattagaa attttattga tagaagtatt

     6721 ttacaaatac aaatacatac taagggtttc ttatatgctc aacacatgag cgaaacccta

     6781 taggaaccct aattccctta tctgggaact actcacacat tattatggag aaactcgagc

     6841 ttgtcgatcg acagatccgg tcggcatcta ctctatttct ttgccctcgg acgagtgctg

     6901 gggcgtcggt ttccactatc ggcgagtact tctacacagc catcggtcca gacggccgcg

     6961 cttctgcggg cgatttgtgt acgcccgaca gtcccggctc cggatcggac gattgcgtcg

     7021 catcgaccct gcgcccaagc tgcatcatcg aaattgccgt caaccaagct ctgatagagt

     7081 tggtcaagac caatgcggag catatacgcc cggagtcgtg gcgatcctgc aagctccgga

     7141 tgcctccgct cgaagtagcg cgtctgctgc tccatacaag ccaaccacgg cctccagaag

     7201 aagatgttgg cgacctcgta ttgggaatcc ccgaacatcg cctcgctcca gtcaatgacc

     7261 gctgttatgc ggccattgtc cgtcaggaca ttgttggagc cgaaatccgc gtgcacgagg

     7321 tgccggactt cggggcagtc ctcggcccaa agcatcagct catcgagagc ctgcgcgacg

     7381 gacgcactga cggtgtcgtc catcacagtt tgccagtgat acacatgggg atcagcaatc

     7441 gcgcatatga aatcacgcca tgtagtgtat tgaccgattc cttgcggtcc gaatgggccg

     7501 aacccgctcg tctggctaag atcggccgca gcgatcgcat ccatagcctc cgcgaccggt

     7561 tgtagaacag cgggcagttc ggtttcaggc aggtcttgca acgtgacacc ctgtgcacgg

     7621 cgggagatgc aataggtcag gctctcgcta aactccccaa tgtcaagcac ttccggaatc

     7681 gggagcgcgg ccgatgcaaa gtgccgataa acataacgat ctttgtagaa accatcggcg

     7741 cagctattta cccgcaggac atatccacgc cctcctacat cgaagctgaa agcacgagat

     7801 tcttcgccct ccgagagctg catcaggtcg gagacgctgt cgaacttttc gatcagaaac

     7861 ttctcgacag acgtcgcggt gagttcaggc tttttcatat ctcattgccc cccgggatct

     7921 gcgaaagctc gagagagata gatttgtaga gagagactgg tgatttcagc gtgtcctctc

     7981 caaatgaaat gaacttcctt atatagagga aggtcttgcg aaggatagtg ggattgtgcg

     8041 tcatccctta cgtcagtgga gatatcacat caatccactt gctttgaaga cgtggttgga

     8101 acgtcttctt tttccacgat gctcctcgtg ggtgggggtc catctttggg accactgtcg

     8161 gcagaggcat cttgaacgat agcctttcct ttatcgcaat gatggcattt gtaggtgcca

     8221 ccttcctttt ctactgtcct tttgatgaag tgacagatag ctgggcaatg gaatccgagg

     8281 aggtttcccg atattaccct ttgttgaaaa gtctcaatag ccctttggtc ttctgagact

     8341 gtatctttga tattcttgga gtagacgaga gtgtcgtgct ccaccatgtt atcacatcaa

     8401 tccacttgct ttgaagacgt ggttggaacg tcttcttttt ccacgatgct cctcgtgggt

     8461 gggggtccat ctttgggacc actgtcggca gaggcatctt gaacgatagc ctttccttta

     8521 tcgcaatgat ggcatttgta ggtgccacct tccttttcta ctgtcctttt gatgaagtga

     8581 cagatagctg ggcaatggaa tccgaggagg tttcccgata ttaccctttg ttgaaaagtc

     8641 tcaatagccc tttggtcttc tgagactgta tctttgatat tcttggagta gacgagagtg

     8701 tcgtgctcca ccatgttggc aagctgctct agccaatacg caaaccgcct ctccccgcgc

     8761 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg

     8821 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta

     8881 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca

     8941 gctatgacca tgattacg

//