LOCUS       V019260                 6638 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019260
VERSION     V019260
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             671..1318
                     /label="FGF19(NM_005117)"
                     /note="FGF19(NM_005117)"
                     /gene="FGF19"
     CDS             1325..1390
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1414..2002
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2068..2116
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2318..2746
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2760..3089
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3156..3950
                     /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    4126..4247
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4296..4312)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4320..4336
                     /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(4344..4374)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4389..4410
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4698..5286)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5457..6317)
                     /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(6318..6422)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgccacc atgcggagcg ggtgtgtggt ggtccacgta tggatcctgg ccggcctctg
      721 gctggccgtg gccgggcgcc ccctcgcctt ctcggacgcg gggccccacg tgcactacgg
      781 ctggggcgac cccatccgcc tgcggcacct gtacacctcc ggcccccacg ggctctccag
      841 ctgcttcctg cgcatccgtg ccgacggcgt cgtggactgc gcgcggggcc agagcgcgca
      901 cagtttgctg gagatcaagg cagtcgctct gcggaccgtg gccatcaagg gcgtgcacag
      961 cgtgcggtac ctctgcatgg gcgccgacgg caagatgcag gggctgcttc agtactcgga
     1021 ggaagactgt gctttcgagg aggagatccg cccagatggc tacaatgtgt accgatccga
     1081 gaagcaccgc ctcccggtct ccctgagcag tgccaaacag cggcagctgt acaagaacag
     1141 aggctttctt ccactctctc atttcctgcc catgctgccc atggtcccag aggagcctga
     1201 ggacctcagg ggccacttgg aatctgacat gttctcttcg cccctggaga ccgacagcat
     1261 ggacccattt gggcttgtca ccggactgga ggccgtgagg agtcccagct ttgagaagct
     1321 cgaggactac aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga
     1381 cgatgacaag tagtgagggc ccgatatctc gacaatcaac ctctggatta caaaatttgt
     1441 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     1501 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     1561 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     1621 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     1681 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     1741 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     1801 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
     1861 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     1921 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     1981 tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca cggaaggaga
     2041 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacgggt
     2101 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     2161 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     2221 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     2281 gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     2341 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2401 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2461 gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2521 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2581 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2641 acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg gatttcggcc
     2701 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     2761 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2821 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2881 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     2941 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3001 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     3061 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     3121 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3181 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3241 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3301 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     3361 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3421 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     3481 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     3541 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     3601 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     3661 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     3721 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     3781 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     3841 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     3901 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     3961 ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     4021 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4081 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     4141 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4201 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     4261 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4321 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4381 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4441 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4501 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4561 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4621 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4681 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4741 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4801 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4861 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg
     4921 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4981 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5041 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5101 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     5161 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5221 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5281 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5341 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5401 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5461 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5521 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5581 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5641 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5701 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5761 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5821 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5881 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5941 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6001 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6061 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6121 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6181 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6241 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6301 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6361 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6421 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     6481 tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     6541 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     6601 aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
