LOCUS       V043891                 6386 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043891
VERSION     V043891
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..1906
                     /label="Gapdh(NM_008084)"
                     /note="Gapdh(NM_008084)"
                     /gene="Gapdh"
     polyA_signal    1953..2177
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2223..2651
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2665..2994
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3061..3855
                     /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    4029..4150
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4199..4215)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4223..4239
                     /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(4247..4277)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4292..4313
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4601..5186)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5357..6217)
                     /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(6218..6322)
                     /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 gtgaaggtcg gtgtgaacgg atttggccgt attgggcgcc tggtcaccag
      961 ggctgccatt tgcagtggca aagtggagat tgttgccatc aacgacccct tcattgacct
     1021 caactacatg gtctacatgt tccagtatga ctccactcac ggcaaattca acggcacagt
     1081 caaggccgag aatgggaagc ttgtcatcaa cgggaagccc atcaccatct tccaggagcg
     1141 agaccccact aacatcaaat ggggtgaggc cggtgctgag tatgtcgtgg agtctactgg
     1201 tgtcttcacc accatggaga aggccggggc ccacttgaag ggtggagcca aaagggtcat
     1261 catctccgcc ccttctgccg atgcccccat gtttgtgatg ggtgtgaacc acgagaaata
     1321 tgacaactca ctcaagattg tcagcaatgc atcctgcacc accaactgct tagcccccct
     1381 ggccaaggtc atccatgaca actttggcat tgtggaaggg ctcatgacca cagtccatgc
     1441 catcactgcc acccagaaga ctgtggatgg cccctctgga aagctgtggc gtgatggccg
     1501 tggggctgcc cagaacatca tccctgcatc cactggtgct gccaaggctg tgggcaaggt
     1561 catcccagag ctgaacggga agctcactgg catggccttc cgtgttccta cccccaatgt
     1621 gtccgtcgtg gatctgacgt gccgcctgga gaaacctgcc aagtatgatg acatcaagaa
     1681 ggtggtgaag caggcatctg agggcccact gaagggcatc ttgggctaca ctgaggacca
     1741 ggttgtctcc tgcgacttca acagcaactc ccactcttcc accttcgatg ccggggctgg
     1801 cattgctctc aatgacaact ttgtcaagct catttcctgg tatgacaatg aatacggcta
     1861 cagcaacagg gtggtggacc tcatggccta catggcctcc aaggagtaac tcgagtctag
     1921 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     1981 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2041 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     2101 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     2161 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     2221 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2281 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2341 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2401 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     2461 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     2521 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     2581 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     2641 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     2701 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     2761 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2821 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     2881 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     2941 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     3001 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     3061 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     3121 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     3181 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     3241 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     3301 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     3361 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     3421 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     3481 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     3541 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     3601 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     3661 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     3721 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     3781 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     3841 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     3901 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     3961 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4021 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4081 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4141 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     4201 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4261 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4321 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4381 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4441 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     4501 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4561 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4621 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4681 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4741 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4801 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4861 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4921 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4981 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5041 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5101 gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga
     5161 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5221 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5281 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5341 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5401 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5461 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5521 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5581 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5641 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5701 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5761 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5821 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5881 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5941 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6001 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6061 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6121 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6181 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6241 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6301 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     6361 cgggagatct cccgatcccc tatggt
//
