LOCUS       V032070                 8012 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032070
VERSION     V032070
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..3505
                     /label="FGA(NM_000508)"
                     /note="FGA(NM_000508)"
                     /gene="FGA"
     CDS             3512..3538
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    3579..3803
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3849..4277
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4291..4620
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4687..5481
                     /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    5655..5776
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5825..5841)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5849..5865
                     /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(5873..5903)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5918..5939
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6227..6812)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6983..7843)
                     /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(7844..7948)
                     /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 ttttccatga ggatcgtctg cctggtccta agtgtggtgg gcacagcatg
      961 gactgcagat agtggtgaag gtgactttct agctgaagga ggaggcgtgc gtggcccaag
     1021 ggttgtggaa agacatcaat ctgcctgcaa agattcagac tggcccttct gctctgatga
     1081 agactggaac tacaaatgcc cttctggctg caggatgaaa gggttgattg atgaagtcaa
     1141 tcaagatttt acaaacagaa taaataagct caaaaattca ctatttgaat atcagaagaa
     1201 caataaggat tctcattcgt tgaccactaa tataatggaa attttgagag gcgatttttc
     1261 ctcagccaat aaccgtgata atacctacaa ccgagtgtca gaggatctga gaagcagaat
     1321 tgaagtcctg aagcgcaaag tcatagaaaa agtacagcat atccagcttc tgcagaaaaa
     1381 tgttagagct cagttggttg atatgaaacg actggaggtg gacattgata ttaagatccg
     1441 atcttgtcga gggtcatgca gtagggcttt agctcgtgaa gtagatctga aggactatga
     1501 agatcagcag aagcaacttg aacaggtcat tgccaaagac ttacttccct ctagagatag
     1561 gcaacactta ccactgataa aaatgaaacc agttccagac ttggttcccg gaaattttaa
     1621 gagccagctt cagaaggtac ccccagagtg gaaggcatta acagacatgc cgcagatgag
     1681 aatggagtta gagagacctg gtggaaatga gattactcga ggaggctcca cctcttatgg
     1741 aaccggatca gagacggaaa gccccaggaa ccctagcagt gctggaagct ggaactctgg
     1801 gagctctgga cctggaagta ctggaaaccg aaaccctggg agctctggga ctggagggac
     1861 tgcaacctgg aaacctggga gctctggacc tggaagtact ggaagctgga actctgggag
     1921 ctctggaact ggaagtactg gaaaccaaaa ccctgggagc cctagacctg gtagtaccgg
     1981 aacctggaat cctggcagct ctgaacgcgg aagtgctggg cactggacct ctgagagctc
     2041 tgtatctggt agtactggac aatggcactc tgaatctgga agttttaggc cagatagccc
     2101 aggctctggg aacgcgaggc ctaacaaccc agactggggc acatttgaag aggtgtcagg
     2161 aaatgtaagt ccagggacaa ggagagagta ccacacagaa aaactggtca cttctaaagg
     2221 agataaagag ctcaggactg gtaaagagaa ggtcacctct ggtagcacaa ccaccacgcg
     2281 tcgttcatgc tctaaaaccg ttactaagac tgttattggt cctgatggtc acaaagaagt
     2341 taccaaagaa gtggtgacct ccgaagatgg ttctgactgt cccgaggcaa tggatttagg
     2401 cacattgtct ggcataggta ctctggatgg gttccgccat aggcaccctg atgaagctgc
     2461 cttcttcgac actgcctcaa ctggaaaaac attcccaggt ttcttctcac ctatgttagg
     2521 agagtttgtc agtgagactg agtctagggg ctcagaatct ggcatcttca caaatacaaa
     2581 ggaatccagt tctcatcacc ctgggatagc tgaattccct tcccgtggta aatcttcaag
     2641 ttacagcaaa caatttacta gtagcacgag ttacaacaga ggagactcca catttgaaag
     2701 caagagctat aaaatggcag atgaggccgg aagtgaagcc gatcatgaag gaacacatag
     2761 caccaagaga ggccatgcta aatctcgccc tgtcagagac tgtgatgatg tcctccaaac
     2821 acatccttca ggtacccaaa gtggcatttt caatatcaag ctaccgggat ccagtaagat
     2881 tttttctgtt tattgcgatc aagagaccag tttgggagga tggcttttga tccagcaaag
     2941 aatggatgga tcactgaatt ttaaccggac ctggcaagac tacaagagag gtttcggcag
     3001 cctgaatgac gagggggaag gagaattctg gctaggcaat gactacctcc acttactaac
     3061 ccaaaggggc tctgttctta gggttgaatt agaggactgg gctgggaatg aagcttatgc
     3121 agaatatcac ttccgggtag gctctgaggc tgaaggctat gccctccaag tctcctccta
     3181 tgaaggcact gcgggtgatg ctctgattga gggttccgta gaggaagggg cagagtacac
     3241 ctctcacaac aacatgcagt tcagcacctt tgacagggat gcagaccagt gggaagagaa
     3301 ctgtgcagaa gtctatgggg gaggctggtg gtataataac tgccaagcag ccaatctcaa
     3361 tggaatctac taccctgggg gctcctatga cccaaggaat aacagtcctt atgagattga
     3421 gaatggagtg gtctgggttt cctttagagg ggcagattat tccctcaggg ctgttcgcat
     3481 gaaaattagg ccccttgtga cccaactcga gtacccatac gacgtaccag attacgctta
     3541 gtctagaggg cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     3601 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     3661 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     3721 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     3781 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     3841 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3901 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     3961 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4021 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     4081 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4141 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4201 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     4261 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4321 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4381 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4441 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4501 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     4561 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4621 aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg
     4681 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4741 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4801 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4861 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4921 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4981 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     5041 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     5101 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     5161 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     5221 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     5281 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     5341 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     5401 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     5461 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     5521 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     5581 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5641 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5701 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5761 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     5821 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5881 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5941 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     6001 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     6061 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     6121 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     6181 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6241 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6301 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     6361 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     6421 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     6481 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     6541 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6601 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6661 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6721 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc
     6781 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6841 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     6901 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     6961 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     7021 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     7081 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     7141 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     7201 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7261 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7321 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7381 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7441 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7501 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7561 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     7621 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7681 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7741 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7801 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7861 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7921 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     7981 acggatcggg agatctcccg atcccctatg gt
//
