LOCUS       V029099                 7585 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029099
VERSION     V029099
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             757..2265
                     /label="CYP3A4(NM_017460)"
                     /note="CYP3A4(NM_017460)"
                     /gene="CYP3A4"
     CDS             2272..2337
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2361..2949
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3015..3063
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3265..3693
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3707..4036
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4103..4897
                     /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    5073..5194
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5243..5259)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5267..5283
                     /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(5291..5321)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5336..5357
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5645..6233)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6404..7264)
                     /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(7265..7369)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg gccaccatgg ctctcatccc agacttggcc
      781 atggaaacct ggcttctcct ggctgtcagc ctggtgctcc tctatctata tggaacccat
      841 tcacatggac tttttaagaa gcttggaatt ccagggccca cacctctgcc ttttttggga
      901 aatattttgt cctaccataa gggcttttgt atgtttgaca tggaatgtca taaaaagtat
      961 ggaaaagtgt ggggctttta tgatggtcaa cagcctgtgc tggctatcac agatcctgac
     1021 atgatcaaaa cagtgctagt gaaagaatgt tattctgtct tcacaaaccg gaggcctttt
     1081 ggtccagtgg gatttatgaa aagtgccatc tctatagctg aggatgaaga atggaagaga
     1141 ttacgatcat tgctgtctcc aaccttcacc agtggaaaac tcaaggagat ggtccctatc
     1201 attgcccagt atggagatgt gttggtgaga aatctgaggc gggaagcaga gacaggcaag
     1261 cctgtcacct tgaaagacgt ctttggggcc tacagcatgg atgtgatcac tagcacatca
     1321 tttggagtga acatcgactc tctcaacaat ccacaagacc cctttgtgga aaacaccaag
     1381 aagcttttaa gatttgattt tttggatcca ttctttctct caataacagt ctttccattc
     1441 ctcatcccaa ttcttgaagt attaaatatc tgtgtgtttc caagagaagt tacaaatttt
     1501 ttaagaaaat ctgtaaaaag gatgaaagaa agtcgcctcg aagatacaca aaagcaccga
     1561 gtggatttcc ttcagctgat gattgactct cagaattcaa aagaaactga gtcccacaaa
     1621 gctctgtccg atctggagct cgtggcccaa tcaattatct ttatttttgc tggctatgaa
     1681 accacgagca gtgttctctc cttcattatg tatgaactgg ccactcaccc tgatgtccag
     1741 cagaaactgc aggaggaaat tgatgcagtt ttacccaata aggcaccacc cacctatgat
     1801 actgtgctac agatggagta tcttgacatg gtggtgaatg aaacgctcag attattccca
     1861 attgctatga gacttgagag ggtctgcaaa aaagatgttg agatcaatgg gatgttcatt
     1921 cccaaagggg tggtggtgat gattccaagc tatgctcttc accgtgaccc aaagtactgg
     1981 acagagcctg agaagttcct ccctgaaaga ttcagcaaga agaacaagga caacatagat
     2041 ccttacatat acacaccctt tggaagtgga cccagaaact gcattggcat gaggtttgct
     2101 ctcatgaaca tgaaacttgc tctaatcaga gtccttcaga acttctcctt caaaccttgt
     2161 aaagaaacac agatccccct gaaattaagc ttaggaggac ttcttcaacc agaaaaaccc
     2221 gttgttctaa aggttgagtc aagggatggc accgtaagtg gagccggatc cgactacaaa
     2281 gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag
     2341 tgagggcccg atatctcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     2401 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     2461 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     2521 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     2581 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     2641 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     2701 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga
     2761 cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     2821 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     2881 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     2941 cctccccgcc tggaaacggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     3001 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg
     3061 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     3121 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     3181 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagca gatctgcgca
     3241 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3301 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3361 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3421 gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3481 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3541 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     3601 tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat tggttaaaaa
     3661 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     3721 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3781 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3841 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3901 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3961 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4021 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     4081 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4141 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4201 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4261 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     4321 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4381 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4441 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4501 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4561 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4621 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4681 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4741 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4801 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4861 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa
     4921 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     4981 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5041 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     5101 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5161 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     5221 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     5281 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     5341 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     5401 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5461 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5521 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5581 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5641 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5701 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5761 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5821 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
     5881 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5941 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6001 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6061 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     6121 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6181 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6241 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6301 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6361 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6421 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6481 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6541 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6601 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6661 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6721 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6781 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6841 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6901 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6961 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7021 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7081 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7141 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7201 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7261 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7321 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7381 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtcgactct
     7441 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     7501 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     7561 caattgcatg aagaatctgc ttagg
//
