LOCUS       V029457                 7906 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029457
VERSION     V029457
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             964..3396
                     /label="Postn(NM_015784)"
                     /note="Postn(NM_015784)"
                     /gene="Postn"
     CDS             3403..3468
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3506..3730
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3776..4204
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4218..4547
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4614..5408
                     /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    5582..5703
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5752..5768)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5776..5792
                     /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(5800..5830)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5845..5866
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6154..6739)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6910..7770)
                     /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(7771..7875)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatggttc ctctcctgcc cttatatgct ctgctgctgc tgttcctgtg tgatattaac
     1021 cctgcaaatg ccaacagtta ctatgacaag gtcctggctc acagccgcat caggggtcgg
     1081 gatcagggcc caaacgtctg tgccctccag caaattctgg gcaccaaaaa gaaatacttc
     1141 agctcctgta agaactggta tcaaggtgct atctgcggga agaaaaccac tgtgctatat
     1201 gaatgctgcc ctggctatat gagaatggaa gggatgaaag gctgccccgc agtgatgcct
     1261 attgaccatg tttatggcac gctgggcatt gtgggagcca ctaccactca gcactactcc
     1321 gatgtctcga agctgagaga agagattgaa ggaaaagggt catacacgta cttcgcgccg
     1381 agtaacgagg cttgggagaa cctggattct gacattcgca gaggactgga gaacaatgtc
     1441 aatgttgagc tactgaatgc cttacacagc cacatggtta ataagagaat gttaaccaag
     1501 gacctgaaac acggcatggt tattccttca atgtacaaca atctggggct ttttattaac
     1561 cattatccca atggggttgt cactgtgaac tgtgctcgag tcatccatgg gaaccagatt
     1621 gccacaaatg gtgtcgtcca tgtcattgac cgtgtcctga cacaaattgg tacctccatc
     1681 caagacttcc ttgaagcaga agacgacctt tcatcattta gagcagccgc catcacctct
     1741 gacctcttgg agtcccttgg aagagatggt cacttcacgc tctttgctcc caccaatgaa
     1801 gctttcgaga aactgccacg aggtgtccta gaaaggatca tgggagacaa agtggcttct
     1861 gaagctctca tgaagtacca catcctaaat accctccagt gctctgaggc catcactgga
     1921 ggagccgtgt ttgagaccat ggaaggaaac actattgaga tagggtgcga aggggacagt
     1981 atctccatta acggaatcaa gatggtgaac aagaaagaca ttgtgactaa gaatggtgtc
     2041 atccacctga ttgatgaagt cctcattcct gattctgcca aacaagttat tgagctggct
     2101 ggaaaacagc aaaccacttt caccgacctg gtagcccaat taggcttggc atcctctctg
     2161 aagccagatg gagagtacac cttattagca cctgtgaaca atgcgttctc tgatgacact
     2221 ctgagcatgg accaacgcct tcttaagcta attctgcaaa atcacatatt gaaagtaaaa
     2281 gttggcctta gcgacctcta caatggacag atactggaaa ccattggagg caaacaactc
     2341 cgagtctttg tgtatcggac ggctatctgc atagaaaact catgcatggt gagaggaagc
     2401 aagcagggaa ggaatggtgc cattcacata ttccgagaaa tcatccaacc agcagagaaa
     2461 tccctgcacg acaagctgcg gcaagacaag cgctttagca tcttcctcag cctccttgaa
     2521 gctgcagatt tgaaagatct cctgacacag cccggagatt ggaccttgtt tgcaccaacc
     2581 aatgatgcct tcaagggaat gactagcgaa gaaagggagc ttctgattgg ggataaaaat
     2641 gctctccaaa acatcattct ttatcacctg accccagggg tttatattgg aaagggattc
     2701 gaacccggag tcactaatat cctgaagacc acacagggaa gcaaaatcta tctgaaagga
     2761 gtaaacgaaa cgcttctagt gaatgagttg aagtccaaag aatctgacat catgacgaca
     2821 aatggtgtca tccacgtcgt ggacaaactc ctctatccag cagatattcc agttggaaat
     2881 gatcagctct tggaattact gaacaaactg ataaaataca tccaaatcaa gtttgttcgt
     2941 ggcagcacct tcaaagaaat ccccatgact gtctatagac ctgcaatgac gaagatccaa
     3001 attgaaggtg atcccgactt caggctgatt aaagaaggcg aaacggtgac agaagtgatc
     3061 cacggagagc cagtcattaa aaagtacacc aaaatcatag atggagttcc tgttgaaata
     3121 actgaaaaac agactcggga agaacgaatc attacaggtc ctgagataaa atataccagg
     3181 atttccacag gaggtggaga aacaggagag accttgcaga aattcttgca aaaagaggtc
     3241 tccaaggtca caaagttcat tgaaggtggc gatggtcact tatttgaaga tgaggagatt
     3301 aaaagactgc ttcagggaga cacacctgca aagaagatac cagccaacaa aagggttcaa
     3361 gggcctagaa gacgatcaag agaaggccgt tctcagctcg aggactacaa agaccatgac
     3421 ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta gtgagggccc
     3481 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     3541 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     3601 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     3661 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     3721 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     3781 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3841 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3901 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3961 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     4021 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4081 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     4141 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4201 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     4261 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4321 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4381 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4441 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     4501 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     4561 cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     4621 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     4681 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     4741 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     4801 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     4861 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     4921 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     4981 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     5041 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     5101 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     5161 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     5221 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     5281 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     5341 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     5401 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     5461 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5521 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5581 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5641 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5701 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     5761 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5821 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5881 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5941 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6001 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6061 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6121 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6181 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6241 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6301 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6361 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6421 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6481 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6541 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6601 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6661 atccggcaaa caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg
     6721 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6781 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6841 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6901 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6961 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7021 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7081 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7141 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7201 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7261 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7321 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7381 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7441 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7501 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7561 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7621 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7681 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7741 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7801 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7861 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
