LOCUS       V024642                 7661 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V024642
VERSION     V024642
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             779..2341
                     /label="CYP11A1(NM_000781)"
                     /note="CYP11A1(NM_000781)"
                     /gene="CYP11A1"
     CDS             2348..2413
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2437..3025
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3091..3139
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3341..3769
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3783..4112
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4179..4973
                     /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    5149..5270
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5319..5335)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5343..5359
                     /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(5367..5397)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5412..5433
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5721..6309)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6480..7340)
                     /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(7341..7445)
                     /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 ctcggatcca ctagtccagt gtggtggaat tcgccaccat
      781 gctggccaag ggtcttcccc cacgctcagt cctggtcaaa ggctgccaga cctttctgag
      841 tgcccccagg gaggggctgg ggcgtctcag ggtgcccact ggcgagggag ctggcatctc
      901 cacccgcagt cctcgcccct tcaatgagat cccctctcct ggtgacaatg gctggctaaa
      961 cctgtaccat ttctggaggg agacgggcac acacaaagtc caccttcacc atgtccagaa
     1021 tttccagaag tatggcccga tttacaggga gaagctcggc aacgtggagt cggtttatgt
     1081 catcgaccct gaagatgtgg cccttctctt taagtccgag ggccccaacc cagaacgatt
     1141 cctcatcccg ccctgggtcg cctatcacca gtattaccag agacccatag gagtcctgtt
     1201 gaagaagtcg gcagcctgga agaaagaccg ggtggccctg aaccaggagg tgatggctcc
     1261 agaggccacc aagaactttt tgcccctgtt ggatgcagtg tctcgggact tcgtcagtgt
     1321 cctgcacagg cgcatcaaga aggcgggctc cggaaattac tcgggggaca tcagtgatga
     1381 cctgttccgc tttgcctttg agtccatcac taacgtcatt tttggggagc gccaggggat
     1441 gctggaggaa gtagtgaacc ccgaggccca gcgattcatt gatgccatct accagatgtt
     1501 ccacaccagc gtccccatgc tcaaccttcc cccagacctg ttccgtctgt tcaggaccaa
     1561 gacctggaag gaccatgtgg ctgcatggga cgtgattttc agtaaagctg acatatacac
     1621 ccagaacttc tactgggaat tgagacagaa aggaagtgtt caccacgatt accgtggcat
     1681 cctctacaga ctcctgggag acagcaagat gtccttcgag gacatcaagg ccaacgtcac
     1741 agagatgctg gcaggagggg tggacacgac gtccatgacc ctgcagtggc acttgtatga
     1801 gatggcacgc aacctgaagg tgcaggatat gctgcgggca gaggtcttgg ctgcgcggca
     1861 ccaggcccag ggagacatgg ccacgatgct acagctggtc cccctcctca aagccagcat
     1921 caaggagaca ctaagacttc accccatctc cgtgaccctg cagagatatc ttgtaaatga
     1981 cttggttctt cgagattaca tgattcctgc caagacactg gtgcaagtgg ccatctatgc
     2041 tctgggccga gagcccacct tcttcttcga cccggaaaat tttgacccaa cccgatggct
     2101 gagcaaagac aagaacatca cctacttccg gaacttgggc tttggctggg gtgtgcggca
     2161 gtgtctggga cggcggatcg ctgagctaga gatgaccatc ttcctcatca atatgctgga
     2221 gaacttcaga gttgaaatcc aacacctcag cgatgtgggc accacattca acctcattct
     2281 gatgcctgaa aagcccatct ccttcacctt ctggcccttt aaccaggaag caacccagca
     2341 gctcgaggac tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga
     2401 tgacgatgac aagtagtgag ggcccgatat ctcgacaatc aacctctgga ttacaaaatt
     2461 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     2521 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     2581 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     2641 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     2701 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     2761 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     2821 ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg
     2881 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     2941 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     3001 atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa acacggaagg
     3061 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     3121 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     3181 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     3241 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     3301 atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     3361 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3421 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3481 caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3541 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3601 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3661 acaacactca accctatctc ggtctattct tttgatttat aagggatttt ggggatttcg
     3721 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     3781 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3841 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3901 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3961 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4021 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     4081 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     4141 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4201 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4261 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4321 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     4381 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4441 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4501 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4561 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4621 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4681 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     4741 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4801 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4861 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4921 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4981 tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     5041 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     5101 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     5161 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     5221 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     5281 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     5341 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     5401 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     5461 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5521 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5581 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5641 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5701 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5761 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5821 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5881 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt
     5941 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6001 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6061 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6121 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     6181 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6241 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6301 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6361 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6421 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6481 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6541 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6601 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6661 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6721 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6781 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6841 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6901 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6961 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7021 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7081 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7141 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7201 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7261 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7321 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7381 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7441 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     7501 ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     7561 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     7621 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g
//
