LOCUS       V029497                 8045 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029497
VERSION     V029497
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             744..770
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             774..800
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             804..830
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             840..881
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /label="V5 tag"
     CDS             888..2720
                     /label="Gbp6(NM_194336)"
                     /note="Gbp6(NM_194336)"
                     /gene="Gbp6"
     misc_feature    2821..3409
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3475..3523
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3725..4153
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4167..4496
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4563..5357
                     /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    5533..5654
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5703..5719)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5727..5743
                     /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(5751..5781)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5796..5817
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6105..6693)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6864..7724)
                     /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(7725..7829)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
      661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
      721 actctagagc tagcgccacc atgtacccat acgacgtacc tgattatgca acttatcctt
      781 atgatgtacc tgattatgct agctatcctt atgatgtccc agactacgca ggcggaggtg
      841 gtaagcctat ccctaaccct ctcctcggtc tcgattctac aggatccatg acccaaccac
      901 aaatggctcc catttgcctt gtggaaaacc acaatgaaca gctgtcagtg aaccaggaag
      961 ccatagagat tctggacaag atttctcagc cagtggtagt cgtggctatt gttggattgt
     1021 accgtacagg gaagtcctat ttgatgaact gtttggcggg acagaatcac ggctttcctc
     1081 tgggctccac cgtgcagtct cagaccaagg gcatctggat gtggtgcatg ccacacccca
     1141 ccaagccaga gcacaccctg gtcctcctgg acactgaggg cctgggggat gtggaaaagg
     1201 gtgatcctaa gaacgacttg tggatctttg ccctcagcgt gcttctgagc agcaccttca
     1261 tctacaacag catgatcacc atcaaccacc aggccctgga gcagctgcat tatgtcacag
     1321 aactcacaga gctgatcaga gcaaagtctt ccccaaatcc tgctggaata aagaattcca
     1381 cagagtttgt gagtttcttt ccagactttg tctggactgt tcgggatttc atgctggagc
     1441 tgaagttaaa tggggaagac atcacaagtg atgactacct ggagaatgcc ttgaagctga
     1501 tcccaggtga caaacccaga atgcaagcat ccaattcatg cagggaatgc atcagacttt
     1561 tctttcctaa ccggaagtgt tttgtctttg accggccaac gcatgacaaa gaacttttac
     1621 aaaaacttga ttctatcaca gaagaccaac tggatcctaa gttccaggaa gtaacaaagg
     1681 cttttgtttc ttacatcttc acttatgcca agatcaagac cctaaaagag ggaattaagg
     1741 tcactgggaa tagactaggg attctggtga caacctatgt gaatgccatc aacagtggag
     1801 cagtgccttg tctggatgat gctgtgacaa ctctggccca gcgtgagaac tcagtagctg
     1861 tgcagaaagc agccgaccac tatagtgagc agatggccca gcgactgagg cttcctacag
     1921 aaacgctcca ggagctgctg gatgtgcatg cagcctgcga gaaggaagcc atggctgtct
     1981 tcatggagca ttccttcaag gacgaaaatc agcaattcct gaagaagctg gtggaattaa
     2041 taggagagaa caaagagctt ttcctgtcga agaatgaaga ggcatcaaat aaatactgtc
     2101 aagaagaact ggatcgactt tcaaaggatt ttatggaaaa tatttcaaca ttttttgttc
     2161 cttgtggaca caagctttac atggacaaga gggagaagat tgaacatgac tactggcagg
     2221 ttcccaggaa aggggtgaag gcaagtgaag tcttccagag ctttctgcag tcacaggcct
     2281 tcatcgagag ttccatcttg caggcagata cagcgctcac tgctggggag aaggccattg
     2341 cagaggagcg tgcccagaag gtggcggcag agaaggagca agagctgcta agacagaagc
     2401 agaaggagca gcaggagtat atggaggctc aagagaaaag tcacaaggaa aacctagagc
     2461 aactgagaag gaagctggag caggagagag agcaggacat caaagaccat gatatgatgc
     2521 tgaagaagct aatgaaggat caaaaggctt tccttgagga aggatttaag aagaaagctg
     2581 aagaaatgaa caaagagata cagcaactga gagatgtcat caaggataag aaaagaaaca
     2641 ctgatcgaat taaggaggct ctcttaaatg gattttctac agttcttttt cattaccttg
     2701 tccgttatct aaagcattta tgaattctgc agatatccag cacagtggcg gccgctcgag
     2761 tctagagggc cccttaaggg ttcgatccct accggttagt aatgagtttg atatctcgac
     2821 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     2881 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     2941 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     3001 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     3061 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     3121 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     3181 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc
     3241 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     3301 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     3361 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaaacggg
     3421 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     3481 taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg
     3541 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     3601 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     3661 aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat
     3721 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3781 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3841 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga
     3901 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3961 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4021 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     4081 ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4141 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4201 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4261 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4321 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4381 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     4441 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     4501 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     4561 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4621 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4681 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4741 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4801 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4861 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4921 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4981 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5041 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     5101 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5161 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5221 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5281 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5341 ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc
     5401 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     5461 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     5521 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     5581 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5641 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     5701 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     5761 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     5821 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5881 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5941 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     6001 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     6061 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6121 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6181 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6241 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa
     6301 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6361 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6421 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6481 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6541 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6601 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6661 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6721 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6781 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6841 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6901 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6961 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     7021 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     7081 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     7141 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     7201 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     7261 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     7321 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7381 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7441 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     7501 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     7561 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7621 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7681 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7741 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7801 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     7861 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
     7921 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     7981 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     8041 ttagg
//
