LOCUS       V020106                 6893 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020106
VERSION     V020106
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             932..2347
                     /label="FOXA1(NM_004496)"
                     /note="FOXA1(NM_004496)"
                     /gene="FOXA1"
     CDS             2378..2407
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /transl_table=1
                     /translation="EQKLISEEDL"
                     /label="Myc"
     CDS             2423..2440
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    2490..2714
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2760..3188
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3202..3531
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3598..4392
                     /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    4566..4687
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4736..4752)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4760..4776
                     /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(4784..4814)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4829..4850
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5138..5726)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5897..6757)
                     /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(6758..6862)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaacgg gccctctaga ctcgagccac catgttagga actgtgaaga tggaagggca
      961 tgaaaccagc gactggaaca gctactacgc agacacgcag gaggcctact cctccgtccc
     1021 ggtcagcaac atgaactcag gcctgggctc catgaactcc atgaacacct acatgaccat
     1081 gaacaccatg actacgagcg gcaacatgac cccggcgtcc ttcaacatgt cctatgccaa
     1141 cccgggccta ggggccggcc tgagtcccgg cgcagtagcc ggcatgccgg ggggctcggc
     1201 gggcgccatg aacagcatga ctgcggccgg cgtgacggcc atgggtacgg cgctgagccc
     1261 gagcggcatg ggcgccatgg gtgcgcagca ggcggcctcc atgaatggcc tgggccccta
     1321 cgcggccgcc atgaacccgt gcatgagccc catggcgtac gcgccgtcca acctgggccg
     1381 cagccgcgcg ggcggcggcg gcgacgccaa gacgttcaag cgcagctacc cgcacgccaa
     1441 gccgccctac tcgtacatct cgctcatcac catggccatc cagcaggcgc ccagcaagat
     1501 gctcacgctg agcgagatct accagtggat catggacctc ttcccctatt accggcagaa
     1561 ccagcagcgc tggcagaact ccatccgcca ctcgctgtcc ttcaatgact gcttcgtcaa
     1621 ggtggcacgc tccccggaca agccgggcaa gggctcctac tggacgctgc acccggactc
     1681 cggcaacatg ttcgagaacg gctgctactt gcgccgccag aagcgcttga agtgcgagaa
     1741 gcagccgggg gccggcggcg ggggcgggag cggaagcggg ggcagcggcg ccaagggcgg
     1801 ccctgagagc cgcaaggacc cctctggcgc ctctaacccc agcgccgact cgcccctcca
     1861 tcggggtgtg cacgggaaga ccggccagct agagggcgcg ccggcccccg ggcccgccgc
     1921 cagcccccag actctggacc acagtggggc gacggcgaca gggggcgcct cggagttgaa
     1981 gactccagcc tcctcaactg cgccccccat aagctccggg cccggggcgc tggcctctgt
     2041 gcccgcctct cacccggcac acggcttggc accccacgag tcccagctgc acctgaaagg
     2101 ggacccccac tactccttca accacccgtt ctccatcaac aacctcatgt cctcctcgga
     2161 gcagcagcat aagctggact tcaaggcata cgaacaggca ctgcaatact cgccttacgg
     2221 ctctacgttg cccgccagcc tgcctctagg cagcgcctcg gtgaccacca ggagccccat
     2281 cgagccctca gccctggagc cggcgtacta ccaaggtgtg tattccagac ccgtcctaaa
     2341 cacttccgga tccgagctcg gtaccaagct tgggcccgaa caaaaactca tctcagaaga
     2401 ggatctgaat agcgccgtcg accatcatca tcatcatcat tgagtttaaa cggtctccag
     2461 cttaagttta aaccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     2521 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     2581 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     2641 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     2701 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     2761 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2821 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2881 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2941 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3001 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3061 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3121 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3181 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3241 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3301 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3361 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3421 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     3481 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     3541 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     3601 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3661 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     3721 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     3781 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     3841 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     3901 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     3961 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4021 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4081 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     4141 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4201 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     4261 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4321 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4381 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     4441 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     4501 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     4561 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4621 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4681 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     4741 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4801 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4861 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4921 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4981 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5041 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5101 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5161 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     5221 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     5281 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     5341 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     5401 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5461 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5521 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5581 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5641 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5701 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5761 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5821 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5881 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5941 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6001 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6061 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6121 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6181 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6241 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6301 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6361 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6421 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6481 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6541 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6601 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6661 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6721 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6781 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6841 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
