LOCUS       V025304                 6950 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025304
VERSION     V025304
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..2164
                     /label="BZW2(NM_014038)"
                     /note="BZW2(NM_014038)"
                     /gene="BZW2"
     polyA_signal    2517..2741
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2787..3215
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3229..3558
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3625..4419
                     /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    4593..4714
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4763..4779)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4787..4803
                     /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(4811..4841)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4856..4877
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5165..5750)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5921..6781)
                     /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(6782..6886)
                     /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 aataagcatc agaagccagt gctaacaggc cagcggttca aaactcggaa
      961 aagggatgaa aaagagaaat tcgaacccac agtcttcagg gatacacttg tccaggggct
     1021 taatgaggct ggtgatgacc ttgaagctgt agccaaattt ctggactcta caggctcaag
     1081 attagattat cgtcgctatg cagacacact cttcgatatc ctggtggctg gcagtatgct
     1141 tgcccctgga ggaacgcgca tagatgatgg tgacaagacc aagatgacca accactgtgt
     1201 gttttcagca aatgaagatc atgaaaccat ccgaaactat gctcaggtct tcaataaact
     1261 catcaggaga tataagtatt tggagaaggc atttgaagat gaaatgaaaa agcttctcct
     1321 cttccttaaa gccttttccg aaacagagca gacaaagttg gcgatgctgt cggggattct
     1381 gctgggcaat ggcaccctgc ccgccaccat cctcaccagt ctcttcaccg acagcttagt
     1441 caaagaaggc attgcggcct catttgctgt caagcttttc aaagcatgga tggcagaaaa
     1501 agatgccaac tctgttacct cgtctttgag aaaagccaac ttagacaaga ggctgcttga
     1561 actctttcca gttaacagac agagtgtgga tcattttgct aaatacttca ctgacgcagg
     1621 tcttaaggag ctttccgact tcctccgagt ccagcagtcc ctgggcacca ggaaggaact
     1681 gcagaaggag ctccaggagc gtctttctca ggaatgcccg atcaaggagg tggtgcttta
     1741 tgtcaaagaa gaaatgaaga ggaatgatct tccagaaaca gcagtgattg gtcttctgtg
     1801 gacatgtata atgaacgctg ttgagtggaa caagaaggaa gaacttgttg cagagcaggc
     1861 tctgaagcac ctgaagcaat atgctcccct gctggccgtg ttcagctccc aaggccagtc
     1921 agagctgatc ctcctccaga aggttcagga atactgctac gacaacatcc atttcatgaa
     1981 agcctttcag aagattgtgg ttctctttta taaagctgat gttctgagcg aagaagcaat
     2041 actgaaatgg tataaggaag cacatgttgc taaaggcaaa agtgtttttc ttgaccagat
     2101 gaagaaattt gttgagtggt tacaaaatgc agaagaagaa tccgaatcgg aaggtgagga
     2161 aaatggaggt ggaggatcaa tggccgacaa gcagaagaac ggcatcaagg ccaacttcaa
     2221 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     2281 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     2341 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     2401 cgccgggatc actctcggca tggacgagct gtacaagtaa gaattctgca gatatccagc
     2461 acagtggcgg ccgctcgagt ctagagggcc cgtttaaacc cgctgatcag cctcgactgt
     2521 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     2581 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     2641 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     2701 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     2761 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     2821 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     2881 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     2941 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3001 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3061 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3121 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     3181 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     3241 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3301 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3361 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3421 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3481 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3541 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     3601 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     3661 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3721 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     3781 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     3841 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3901 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     3961 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     4021 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     4081 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     4141 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     4201 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     4261 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4321 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4381 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     4441 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4501 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     4561 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     4621 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4681 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     4741 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     4801 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     4861 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     4921 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     4981 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5041 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5101 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5161 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5221 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5281 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5341 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5401 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5461 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5521 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5581 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5641 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5701 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5761 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5821 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5881 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5941 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6001 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6061 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6121 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6181 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6241 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6301 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6361 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6421 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6481 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6541 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6601 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6661 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6721 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6781 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6841 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6901 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
//
