LOCUS       V046435                 6889 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V046435
VERSION     V046435
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             931..2328
                     /label="VIM(NM_003380)"
                     /note="VIM(NM_003380)"
                     /gene="VIM"
     CDS             2335..2361
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2365..2391
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2395..2421
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    2456..2680
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2726..3154
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3168..3497
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3564..4358
                     /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    4532..4653
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4702..4718)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4726..4742
                     /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(4750..4780)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4795..4816
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5104..5689)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5860..6720)
                     /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(6721..6825)
                     /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 cactagtcca gtgtggtgga attcgccacc atgtccacca ggtccgtgtc ctcgtcctcc
      961 taccgcagga tgttcggcgg cccgggcacc gcgagccggc cgagctccag ccggagctac
     1021 gtgactacgt ccacccgcac ctacagcctg ggcagcgcgc tgcgccccag caccagccgc
     1081 agcctctacg cctcgtcccc gggcggcgtg tatgccacgc gctcctctgc cgtgcgcctg
     1141 cggagcagcg tgcccggggt gcggctcctg caggactcgg tggacttctc gctggccgac
     1201 gccatcaaca ccgagttcaa gaacacccgc accaacgaga aggtggagct gcaggagctg
     1261 aatgaccgct tcgccaacta catcgacaag gtgcgcttcc tggagcagca gaataagatc
     1321 ctgctggccg agctcgagca gctcaagggc caaggcaagt cgcgcctggg ggacctctac
     1381 gaggaggaga tgcgggagct gcgccggcag gtggaccagc taaccaacga caaagcccgc
     1441 gtcgaggtgg agcgcgacaa cctggccgag gacatcatgc gcctccggga gaaattgcag
     1501 gaggagatgc ttcagagaga ggaagccgaa aacaccctgc aatctttcag acaggatgtt
     1561 gacaatgcgt ctctggcacg tcttgacctt gaacgcaaag tggaatcttt gcaagaagag
     1621 attgcctttt tgaagaaact ccacgaagag gaaatccagg agctgcaggc tcagattcag
     1681 gaacagcatg tccaaatcga tgtggatgtt tccaagcctg acctcacggc tgccctgcgt
     1741 gacgtacgtc agcaatatga aagtgtggct gccaagaacc tgcaggaggc agaagaatgg
     1801 tacaaatcca agtttgctga cctctctgag gctgccaacc ggaacaatga cgccctgcgc
     1861 caggcaaagc aggagtccac tgagtaccgg agacaggtgc agtccctcac ctgtgaagtg
     1921 gatgccctta aaggaaccaa tgagtccctg gaacgccaga tgcgtgaaat ggaagagaac
     1981 tttgccgttg aagctgctaa ctaccaagac actattggcc gcctgcagga tgagattcag
     2041 aatatgaagg aggaaatggc tcgtcacctt cgtgaatacc aagacctgct caatgttaag
     2101 atggcccttg acattgagat tgccacctac aggaagctgc tggaaggcga ggagagcagg
     2161 atttctctgc ctcttccaaa cttttcctcc ctgaacctga gggaaactaa tctggattca
     2221 ctccctctgg ttgataccca ctcaaaaagg acacttctga ttaagacggt tgaaactaga
     2281 gatggacagg ttatcaacga aacttctcag catcacgatg accttgaact cgagtaccca
     2341 tacgacgtac ctgattatgc aacttatcct tatgatgtac ctgattatgc tagctatcct
     2401 tatgatgtcc cagactacgc atgagggccc gtttaaaccc gctgatcagc ctcgactgtg
     2461 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     2521 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     2581 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     2641 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     2701 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2761 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     2821 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2881 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2941 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3001 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3061 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3121 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     3181 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3241 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3301 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3361 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3421 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3481 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     3541 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     3601 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     3661 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     3721 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     3781 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     3841 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     3901 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     3961 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4021 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4081 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4141 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4201 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4261 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4321 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     4381 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4441 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4501 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     4561 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4621 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     4681 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4741 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     4801 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4861 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4921 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4981 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5041 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5101 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5161 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5221 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5281 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5341 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5401 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5461 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5521 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5581 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5641 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5701 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5761 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5821 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5881 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5941 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6001 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6061 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6121 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6181 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6241 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6301 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6361 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6421 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6481 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6541 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6601 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6661 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6721 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6781 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6841 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggt
//
