LOCUS       V023132                 7958 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023132
VERSION     V023132
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             941..3448
                     /label="POSTN(NM_006475)"
                     /note="POSTN(NM_006475)"
                     /gene="POSTN"
     CDS             3455..3520
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3558..3782
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3828..4256
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4270..4599
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4666..5460
                     /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    5634..5755
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5804..5820)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5828..5844
                     /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(5852..5882)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5897..5918
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6206..6791)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6962..7822)
                     /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(7823..7927)
                     /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 atccgccacc atgattccct ttttacccat
      961 gttttctcta ctattgctgc ttattgttaa ccctataaac gccaacaatc attatgacaa
     1021 gatcttggct catagtcgta tcaggggtcg ggaccaaggc ccaaatgtct gtgcccttca
     1081 acagattttg ggcaccaaaa agaaatactt cagcacttgt aagaactggt ataaaaagtc
     1141 catctgtgga cagaaaacga ctgtgttata tgaatgttgc cctggttata tgagaatgga
     1201 aggaatgaaa ggctgcccag cagttttgcc cattgaccat gtttatggca ctctgggcat
     1261 cgtgggagcc accacaacgc agcgctattc tgacgcctca aaactgaggg aggagatcga
     1321 gggaaaggga tccttcactt actttgcacc gagtaatgag gcttgggaca acttggattc
     1381 tgatatccgt agaggtttgg agagcaacgt gaatgttgaa ttactgaatg ctttacatag
     1441 tcacatgatt aataagagaa tgttgaccaa ggacttaaaa aatggcatga ttattccttc
     1501 aatgtataac aatttggggc ttttcattaa ccattatcct aatggggttg tcactgttaa
     1561 ttgtgctcga atcatccatg ggaaccagat tgcaacaaat ggtgttgtcc atgtcattga
     1621 ccgtgtgctt acacaaattg gtacctcaat tcaagacttc attgaagcag aagatgacct
     1681 ttcatctttt agagcagctg ccatcacatc ggacatattg gaggcccttg gaagagacgg
     1741 tcacttcaca ctctttgctc ccaccaatga ggcttttgag aaacttccac gaggtgtcct
     1801 agaaaggatc atgggagaca aagtggcttc cgaagctctt atgaagtacc acatcttaaa
     1861 tactctccag tgttctgagt ctattatggg aggagcagtc tttgagacgc tggaaggaaa
     1921 tacaattgag ataggatgtg acggtgacag tataacagta aatggaatca aaatggtgaa
     1981 caaaaaggat attgtgacaa ataatggtgt gatccatttg attgatcagg tcctaattcc
     2041 tgattctgcc aaacaagtta ttgagctggc tggaaaacag caaaccacct tcacggatct
     2101 tgtggcccaa ttaggcttgg catctgctct gaggccagat ggagaataca ctttgctggc
     2161 acctgtgaat aatgcatttt ctgatgatac tctcagcatg gatcagcgcc tccttaaatt
     2221 aattctgcag aatcacatat tgaaagtaaa agttggcctt aatgagcttt acaacgggca
     2281 aatactggaa accatcggag gcaaacagct cagagtcttc gtatatcgta cagctgtctg
     2341 cattgaaaat tcatgcatgg agaaagggag taagcaaggg agaaacggtg cgattcacat
     2401 attccgcgag atcatcaagc cagcagagaa atccctccat gaaaagttaa aacaagataa
     2461 gcgctttagc accttcctca gcctacttga agctgcagac ttgaaagagc tcctgacaca
     2521 acctggagac tggacattat ttgtgccaac caatgatgct tttaagggaa tgactagtga
     2581 agaaaaagaa attctgatac gggacaaaaa tgctcttcaa aacatcattc tttatcacct
     2641 gacaccagga gttttcattg gaaaaggatt tgaacctggt gttactaaca ttttaaagac
     2701 cacacaagga agcaaaatct ttctgaaaga agtaaatgat acacttctgg tgaatgaatt
     2761 gaaatcaaaa gaatctgaca tcatgacaac aaatggtgta attcatgttg tagataaact
     2821 cctctatcca gcagacacac ctgttggaaa tgatcaactg ctggaaatac ttaataaatt
     2881 aatcaaatac atccaaatta agtttgttcg tggtagcacc ttcaaagaaa tccccgtgac
     2941 tgtctataca actaaaatta taaccaaagt tgtggaacca aaaattaaag tgattgaagg
     3001 cagtcttcag cctattatca aaactgaagg acccacacta acaaaagtca aaattgaagg
     3061 tgaacctgaa ttcagactga ttaaagaagg tgaaacaata actgaagtga tccatggaga
     3121 gccaattatt aaaaaataca ccaaaatcat tgatggagtg cctgtggaaa taactgaaaa
     3181 agagacacga gaagaacgaa tcattacagg tcctgaaata aaatacacta ggatttctac
     3241 tggaggtgga gaaacagaag aaactctgaa gaaattgtta caagaagagg tcaccaaggt
     3301 caccaaattc attgaaggtg gtgatggtca tttatttgaa gatgaagaaa ttaaaagact
     3361 gcttcaggga gacacacccg tgaggaagtt gcaagccaac aaaaaagttc aaggatctag
     3421 aagacgatta agggaaggtc gttctcagct cgaggactac aaagaccatg acggtgatta
     3481 taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc ccgtttaaac
     3541 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3601 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3661 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3721 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     3781 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     3841 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3901 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3961 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4021 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4081 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4141 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     4201 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     4261 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4321 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4381 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4441 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4501 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     4561 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4621 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4681 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4741 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4801 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4861 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4921 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4981 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     5041 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     5101 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     5161 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     5221 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     5281 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5341 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5401 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5461 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     5521 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     5581 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     5641 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5701 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5761 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     5821 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5881 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5941 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     6001 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     6061 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     6121 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     6181 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6241 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6301 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6361 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6421 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6481 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6541 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6601 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6661 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6721 aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6781 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6841 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6901 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6961 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     7021 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     7081 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7141 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7201 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7261 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7321 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7381 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7441 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7501 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7561 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7621 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7681 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7741 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7801 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7861 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7921 ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
