LOCUS       V025596                 9960 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025596
VERSION     V025596
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     primer_bind     141..157
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     enhancer        318..697
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        698..901
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             931..996
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             1000..4665
                     /label="VCPIP1(NM_025054)"
                     /note="VCPIP1(NM_025054)"
                     /gene="VCPIP1"
     polyA_signal    4719..5341
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     promoter        5383..5680
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5734..6528
                     /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    7180..7314
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     promoter        complement(7320..7338)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(7352..7368)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7376..7392
                     /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(7400..7430)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7445..7466
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7754..8342)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8513..9373)
                     /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(9374..9478)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      9505..9933
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa
      961 gatcatgaca tcgattacaa ggatgacgat gacaagctta tgtctcagcc gccgccgccg
     1021 ccgcctccgt tgccgccgcc acctcctccc cctgaggctc cacagactcc gtcgtccttg
     1081 gcgtcggcgg ctgcttcggg ggggcttttg aagcggagag accggagaat cctttccggg
     1141 agctgcccgg atccgaagtg tcaggcgcgt ctatttttcc cggcctccgg ttctgtcagc
     1201 atcgagtgta ccgagtgcgg ccagcggcac gagcagcaac agctgctggg ggttgaggag
     1261 gtgaccgacc cggacgtagt gctacacaac ctgctgcgga acgcgctgct cggggttacg
     1321 ggggcaccca agaagaacac ggaactggta aaggtgatgg gcctttccaa ctatcactgc
     1381 aaattgttgt cgcccatatt agctcgctat ggaatggaca aacagacagg ccgggccaag
     1441 cttctccggg acatgaacca gggcgaactg ttcgattgcg ccttactggg tgaccgcgcc
     1501 ttcctcatag aaccagagca tgttaacact gtgggctatg gcaaggaccg ctccggaagc
     1561 ctcctgtatt tgcatgacac tctggaggac attaagcggg ccaataaaag ccaggaatgt
     1621 ctcattccag tgcatgtgga cggggatgga cactgcttgg tgcatgctgt gtctcgggct
     1681 ctagtaggcc gagagctctt ctggcatgcc ttaagagaga atcttaaaca gcactttcag
     1741 cagcacctgg cccgatatca agctctgttc catgacttca ttgatgctgc tgagtgggag
     1801 gacattatca atgagtgtga ccctctgttt gtaccacctg agggtgttcc cttgggcctg
     1861 aggaatatcc acatatttgg tcttgccaat gtgctacatc gtcctattat tctgttagat
     1921 tccctcagtg gcatgagaag ctctggtgat tattcagcca cctttctacc tgggctcatc
     1981 cctgcagaga agtgcactgg gaaagatggt catttgaaca aaccaatctg tattgcatgg
     2041 agcagctccg gtagaaacca ttatatcccc ttggtaggca taaaaggggc tgctttgccc
     2101 aaactgccta tgaatttgct tcctaaagca tggggtgtgc ctcaggacct tattaaaaag
     2161 tacataaaac ttgaagagga tggtggttgt gttattggag gtgacagaag tttgcaagat
     2221 aaatacttac ttaggcttgt tgctgctatg gaagaagtct ttatggacaa acatggtatc
     2281 catcctagtt tggttgctga tgtccatcag tatttctaca gaaggactgg agtgatagga
     2341 gttcagcctg aggaagtcac agcagctgct aaaaaagcag taatggataa tcgccttcac
     2401 aaatgtttgc tctgtggtgc cctttctgaa cttcatgttc ctccagagtg gttggctcct
     2461 ggagggaaat tgtataacct ggcaaaaagt actcatggac agctgaggac tgacaaaaat
     2521 tacagctttc ccttgaacaa tttggtttgc tcatatgatt cagtgaaaga tgttctggta
     2581 ccagactatg gaatgagtaa cctaacagct tgtaattggt gccatggcac atctgtgcga
     2641 aaggtcagag gagatgggtc tattgtgtat ttggatggag acagaactaa ttctaggtcc
     2701 actggtggca aatgtggttg tggattcaaa cacttttggg atggtaagga gtatgacaat
     2761 ctaccagaag ctttccctat tactttagaa tggggtggaa gagtggtcag agaaacagta
     2821 tattggttcc agtatgaaag tgattcatct ttgaatagta atgtttacga tgttgcaatg
     2881 aaacttgtta ccaagcactt tccaggtgaa tttgggagtg aaatcctagt tcagaaagtt
     2941 gtccacacta tattgcatca gactgccaaa aagaatcccg atgattatac tcctgtaaat
     3001 atagatggtg ctcacgccca aagagttgga gatgttcaag gacaagaatc agagtctcag
     3061 ctcccaacta aaattattct tactggacag aaaacaaaaa ctttgcacaa ggaggagtta
     3121 aacatgagta aaactgaaag aactattcaa cagaatatta cggaacaggc ttctgtaatg
     3181 cagaaacgga aaacagagaa gttaaaacaa gaacaaaaag ggcaacccag gactgtttct
     3241 cccagtacca ttcgtgatgg tccatcctct gcacctgcta cacctaccaa ggctccctat
     3301 tcaccgacaa cttctaagga gaagaagatc cgaatcacaa ctaatgatgg acgacagtcc
     3361 atggttaccc ttaagtcttc aacaaccttt tttgaacttc aggaaagtat agccagagaa
     3421 ttcaacattc ctccatattt acagtgtatt cgatacgggt ttcctcctaa agagttaatg
     3481 ccaccacagg caggaatgga aaaggaacca gttcctttac agcatggcga cagaattaca
     3541 atagaaattc taaaaagtaa agctgaaggt ggtcagtctg ctgcagcaca ctcagcccac
     3601 actgtgaaac aagaagatat tgctgttact ggtaaactgt catctaagga acttcaggag
     3661 caagctgaaa aagaaatgta ctccttgtgt cttttagcaa cattaatggg agaagatgta
     3721 tggtcttatg caaagggact tcctcacatg tttcagcagg gtggtgtatt ctacagtatt
     3781 atgaagaaaa ccatgggtat ggctgatggc aagcattgta cttttccaca tctgcctggc
     3841 aaaacctttg tctataatgc ttctgaagat agactggaat tgtgtgtgga tgctgcagga
     3901 catttcccca ttggtcctga tgttgaagat ttagttaaag aggctgtaag tcaggttcga
     3961 gcagaggcta ctacaagaag tagggaatca agtccctcac atgggctatt aaaactaggt
     4021 agtggtggag tagtgaaaaa gaaatctgag caacttcata acgtaactgc ctttcaggga
     4081 aaagggcatt ctttaggaac tgcatctggt aacccacacc ttgatccaag agctagggaa
     4141 acttcagttg taagaaagca taatacaggg acagacttta gtaatagttc cactaaaaca
     4201 gagccttctg tattcacagc ttcttctagt aatagtgagc ttattcgaat agctcctgga
     4261 gtagtaacaa tgagagacgg caggcagctt gatcctgatt tggttgaggc ccagcgaaaa
     4321 aaattgcagg aaatggtttc ttctattcag gcttcaatgg acaggcacct tcgggatcaa
     4381 agtacagagc agtcaccatc tgatcttcct caaaggaaaa cagaagttgt gagttcttct
     4441 gcaaagtctg ggagtcttca gactggtttg cctgaatctt ttcctttaac tggtggtact
     4501 gaaaatttga atacagaaac aactgatggc tgtgtagcag atgcactggg agcagccttt
     4561 gccacaaggt caaaagcaca aaggggaaat tccgtggagg agcttgaaga gatggatagt
     4621 caagatgctg agatgactaa cacaactgag ccaatggatc actcttgaga attcatcgat
     4681 agatctgata tcggtaccag tcgactctag aggatcccgg gtggcatccc tgtgacccct
     4741 ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa
     4801 taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga
     4861 ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct
     4921 attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct
     4981 gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga
     5041 ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc
     5101 tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat
     5161 tacaggcgtg aaccactgct cccttccctg tccttctgat tttaaaataa ctataccagc
     5221 aggaggacgt ccagacacag cataggctac ctggccatgc ccaaccggtg ggacatttga
     5281 gttgcttgct tggcactgtc ctctcatgcg ttgggtccac tcagtagatg cctgttgaat
     5341 tgggtacgcg gccagcttgg ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     5401 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     5461 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     5521 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     5581 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     5641 cctctgagct attccagaag tagtgaggag gcttttttgg aggaattgat cagcttggga
     5701 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     5761 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     5821 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     5881 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     5941 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     6001 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     6061 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     6121 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     6181 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     6241 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     6301 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     6361 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     6421 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     6481 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     6541 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     6601 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     6661 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc cgggctcgat cccctcgcga
     6721 gttggttcag ctgctgcctg aggctggacg acctcgcgga gttctaccgg cagtgcaaat
     6781 ccgtcggcat ccaggaaacc agcagcggct atccgcgcat ccatgccccc gaactgcagg
     6841 agtggggagg cacgatggcc gctttggtcg acccggacgg gacgctcctg cgcctgatac
     6901 agaacgaatt gcttgcaggc atctcatgag tgtgtcttcc cgttttccgc ctgaggtcac
     6961 tgcgtggatg gagcgctggc gcctgctgcg cgacggcgag ctgctcacca cccactcgcc
     7021 aagctggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gccgatcata
     7081 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     7141 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     7201 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     7261 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcaattcc
     7321 ctatagtgag tcgtattaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt
     7381 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     7441 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     7501 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     7561 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     7621 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     7681 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     7741 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     7801 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     7861 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     7921 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     7981 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     8041 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     8101 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     8161 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     8221 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     8281 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     8341 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     8401 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     8461 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     8521 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     8581 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     8641 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     8701 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     8761 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     8821 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     8881 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     8941 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     9001 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     9061 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     9121 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     9181 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     9241 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     9301 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     9361 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     9421 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     9481 catttccccg aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     9541 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     9601 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     9661 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     9721 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     9781 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     9841 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     9901 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
//
