LOCUS       V040017                 7922 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040017
VERSION     V040017
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             908..3415
                     /label="POSTN(NM_006475)"
                     /note="POSTN(NM_006475)"
                     /gene="POSTN"
     CDS             3425..3442
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    3489..3713
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3759..4187
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4201..4530
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4597..5391
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5565..5686
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5735..5751)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5759..5775
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(5783..5813)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5828..5849
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6137..6722)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6893..7753)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(7754..7858)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg attccctttt tacccatgtt ttctctacta ttgctgctta ttgttaaccc
      961 tataaacgcc aacaatcatt atgacaagat cttggctcat agtcgtatca ggggtcggga
     1021 ccaaggccca aatgtctgtg cccttcaaca gattttgggc accaaaaaga aatacttcag
     1081 cacttgtaag aactggtata aaaagtccat ctgtggacag aaaacgactg tgttatatga
     1141 atgttgccct ggttatatga gaatggaagg aatgaaaggc tgcccagcag ttttgcccat
     1201 tgaccatgtt tatggcactc tgggcatcgt gggagccacc acaacgcagc gctattctga
     1261 cgcctcaaaa ctgagggagg agatcgaggg aaagggatcc ttcacttact ttgcaccgag
     1321 taatgaggct tgggacaact tggattctga tatccgtaga ggtttggaga gcaacgtgaa
     1381 tgttgaatta ctgaatgctt tacatagtca catgattaat aagagaatgt tgaccaagga
     1441 cttaaaaaat ggcatgatta ttccttcaat gtataacaat ttggggcttt tcattaacca
     1501 ttatcctaat ggggttgtca ctgttaattg tgctcgaatc atccatggga accagattgc
     1561 aacaaatggt gttgtccatg tcattgaccg tgtgcttaca caaattggta cctcaattca
     1621 agacttcatt gaagcagaag atgacctttc atcttttaga gcagctgcca tcacatcgga
     1681 catattggag gcccttggaa gagacggtca cttcacactc tttgctccca ccaatgaggc
     1741 ttttgagaaa cttccacgag gtgtcctaga aaggatcatg ggagacaaag tggcttccga
     1801 agctcttatg aagtaccaca tcttaaatac tctccagtgt tctgagtcta ttatgggagg
     1861 agcagtcttt gagacgctgg aaggaaatac aattgagata ggatgtgacg gtgacagtat
     1921 aacagtaaat ggaatcaaaa tggtgaacaa aaaggatatt gtgacaaata atggtgtgat
     1981 ccatttgatt gatcaggtcc taattcctga ttctgccaaa caagttattg agctggctgg
     2041 aaaacagcaa accaccttca cggatcttgt ggcccaatta ggcttggcat ctgctctgag
     2101 gccagatgga gaatacactt tgctggcacc tgtgaataat gcattttctg atgatactct
     2161 cagcatggat cagcgcctcc ttaaattaat tctgcagaat cacatattga aagtaaaagt
     2221 tggccttaat gagctttaca acgggcaaat actggaaacc atcggaggca aacagctcag
     2281 agtcttcgta tatcgtacag ctgtctgcat tgaaaattca tgcatggaga aagggagtaa
     2341 gcaagggaga aacggtgcga ttcacatatt ccgcgagatc atcaagccag cagagaaatc
     2401 cctccatgaa aagttaaaac aagataagcg ctttagcacc ttcctcagcc tacttgaagc
     2461 tgcagacttg aaagagctcc tgacacaacc tggagactgg acattatttg tgccaaccaa
     2521 tgatgctttt aagggaatga ctagtgaaga aaaagaaatt ctgatacggg acaaaaatgc
     2581 tcttcaaaac atcattcttt atcacctgac accaggagtt ttcattggaa aaggatttga
     2641 acctggtgtt actaacattt taaagaccac acaaggaagc aaaatctttc tgaaagaagt
     2701 aaatgataca cttctggtga atgaattgaa atcaaaagaa tctgacatca tgacaacaaa
     2761 tggtgtaatt catgttgtag ataaactcct ctatccagca gacacacctg ttggaaatga
     2821 tcaactgctg gaaatactta ataaattaat caaatacatc caaattaagt ttgttcgtgg
     2881 tagcaccttc aaagaaatcc ccgtgactgt ctatacaact aaaattataa ccaaagttgt
     2941 ggaaccaaaa attaaagtga ttgaaggcag tcttcagcct attatcaaaa ctgaaggacc
     3001 cacactaaca aaagtcaaaa ttgaaggtga acctgaattc agactgatta aagaaggtga
     3061 aacaataact gaagtgatcc atggagagcc aattattaaa aaatacacca aaatcattga
     3121 tggagtgcct gtggaaataa ctgaaaaaga gacacgagaa gaacgaatca ttacaggtcc
     3181 tgaaataaaa tacactagga tttctactgg aggtggagaa acagaagaaa ctctgaagaa
     3241 attgttacaa gaagaggtca ccaaggtcac caaattcatt gaaggtggtg atggtcattt
     3301 atttgaagat gaagaaatta aaagactgct tcagggagac acacccgtga ggaagttgca
     3361 agccaacaaa aaagttcaag gatctagaag acgattaagg gaaggtcgtt ctcaggcggc
     3421 cgcacatcac catcatcacc attgactcga gtctagaggg cccgtttaaa cccgctgatc
     3481 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3541 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     3601 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     3661 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     3721 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     3781 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3841 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3901 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3961 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     4021 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4081 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     4141 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     4201 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4261 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4321 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4381 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4441 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     4501 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     4561 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4621 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4681 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4741 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     4801 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4861 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4921 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4981 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     5041 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     5101 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     5161 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5221 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5281 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5341 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5401 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5461 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5521 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     5581 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5641 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     5701 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     5761 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     5821 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5881 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5941 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6001 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6061 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6121 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6181 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6241 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6301 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6361 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6421 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6481 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6541 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     6601 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6661 ccgctggtag cggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6721 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6781 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6841 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6901 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6961 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7021 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7081 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7141 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7201 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7261 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7321 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7381 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7441 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7501 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7561 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7621 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7681 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7741 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7801 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7861 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     7921 gt
//
