LOCUS       V021672                 7535 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021672
VERSION     V021672
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             683..2182
                     /label="Cyp8b1(NM_010012)"
                     /note="Cyp8b1(NM_010012)"
                     /gene="Cyp8b1"
     CDS             2222..2287
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2311..2899
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2965..3013
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3215..3643
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3657..3986
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4053..4847
                     /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    5023..5144
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5193..5209)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5217..5233
                     /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(5241..5271)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5286..5307
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5595..6183)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6354..7214)
                     /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(7215..7319)
                     /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 taccgagctc ggatccgcca ccatgacgct gtggtgtaca gtgctaggag ccctgctcac
      721 ggttgtggga tgcctgtgtc tgtccttgct gctcaggcac cgcaggccct gggagccacc
      781 tctggacaag ggttttgtgc cctggctggg tcactccatg gctttccgga agaatatgtt
      841 tgaattcttg aaggggatgc gggccaagca cggggatgtc ttcacggtgc agctgggagg
      901 ccagtacttc acctttgtca tggaccccct gtcctttggc cccatcatta agaacacaga
      961 gaaagcgctg gacttccagt cctatgcaaa agaactggtg ctcaaggtgt ttgggtacca
     1021 gtctgtagat ggggatcaca ggatgatcca cttagctagt accaagcatc tgatggggca
     1081 aggcttggag gaactcaacc aggccatgct ggacagccta tccttggtga tgctagggcc
     1141 taaaggttcg agtctgggtg ccagctcctg gtgtgaagat ggcctctttc acttctgcta
     1201 cagaatcctg tttaaggctg gcttcctgag cttattcggc tacaccaagg acaagcagca
     1261 agacctggat gaggccgatg agctgttcag gaagttccgt cgatttgact ttctcttccc
     1321 caggtttgtc tactccctac tggggccccg ggagtgggtg gaagtgagcc agttgcagcg
     1381 tctcttccat cagagactct ctgtgaagca aaacctggaa aaagacggca taagctgttg
     1441 gttaggctac atgcttcagt ttctgaggga gcaaggaata gcttcatcca tgcaggacaa
     1501 atttaacttt atgatgctct gggcctccca ggggaacacg ggacccacct gtttctgggt
     1561 cctcttattc ctgctgaagc accaggatgc catgaaggct gtgcgtgagg aagccaccag
     1621 ggtcatgggt aaggccaggc tggaagccaa gaagtcgttc actttcactc ccagtgccct
     1681 gaaacacact ccagtgttgg acagcgtgat ggaggagagt ctgcgtctct gtgccacccc
     1741 caccctcctc agggtggtac aggaggatta tgtcctgaag atggccagcg gacaagagta
     1801 ccagatccgt cgcggagata aggtggctct cttcccctat ctctcagtac acatggaccc
     1861 cgacatccac cctgagccca cagccttcaa gtatgatcgg ttcctcaacc cggatggcac
     1921 ccggaaagtg gacttctaca agtcgggcaa gaagatccac cactacagca tgccttgggg
     1981 ctcgggtgtt tccaagtgcc ccggcaggtt ctttgccctc agcgagatga agacctttgt
     2041 ccttcttatg attatgtact tcgacttcaa gctggtcgac cccgacatac ctgtgccccc
     2101 cattgaccca aggcgctggg gctttggcac ctcacagcct agccatgagg tacgcttcct
     2161 ctatcgcctg aagcccgtgc aggaattctg cagatatcca gcacagtggc ggccgctcga
     2221 ggactacaaa gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga
     2281 tgacaagtag tgagggcccg atatctcgac aatcaacctc tggattacaa aatttgtgaa
     2341 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     2401 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     2461 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     2521 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     2581 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     2641 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     2701 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     2761 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     2821 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     2881 ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg aaggagacaa
     2941 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt
     3001 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     3061 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     3121 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagca
     3181 gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     3241 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3301 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3361 ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3421 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3481 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3541 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat
     3601 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     3661 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3721 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3781 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3841 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3901 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     3961 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     4021 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4081 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4141 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4201 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     4261 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4321 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4381 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4441 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4501 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4561 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     4621 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     4681 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     4741 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     4801 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     4861 gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     4921 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     4981 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     5041 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     5101 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     5161 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     5221 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     5281 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5341 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5401 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5461 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5521 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5581 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5641 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     5701 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     5761 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     5821 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5881 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5941 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6001 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     6061 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6121 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6181 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6241 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6301 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6361 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6421 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6481 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6541 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6601 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6661 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     6721 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     6781 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     6841 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     6901 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6961 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7021 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7081 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7141 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7201 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7261 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7321 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
     7381 ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
     7441 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
     7501 gcttgaccga caattgcatg aagaatctgc ttagg
//
