LOCUS       V018961                 8441 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018961
VERSION     V018961
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..3097
                     /label="Ahr(NM_013464)"
                     /note="Ahr(NM_013464)"
                     /gene="Ahr"
     CDS             3104..3193
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    3217..3805
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3871..3919
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4121..4549
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4563..4892
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4959..5753
                     /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    5929..6050
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6099..6115)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6123..6139
                     /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(6147..6177)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6192..6213
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6501..7089)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7260..8120)
                     /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(8121..8225)
                     /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 ccatgagcag cggcgccaac atcacctatg ccagccgcaa
      721 gcggcgcaag ccggtgcaga aaacagtaaa gcccatcccc gctgaaggaa ttaagtcaaa
      781 tccttctaag cgacacagag accggctgaa cacagagtta gaccgcctgg ccagcctgct
      841 gcccttcccg caagatgtta ttaataagct ggacaaactc tctgttctta ggctcagcgt
      901 cagctacctg agggccaaga gcttctttga tgttgcatta aagtccaccc ctgctgacag
      961 aaatggaggc caggaccagt gtagagcaca aatcagagac tggcaggatt tgcaagaagg
     1021 agagttcttg ttacaggcgc tgaatggctt tgtgctggtt gtcacagcag atgccttggt
     1081 cttctatgct tcctccacta tccaagatta cctgggcttt cagcagtctg atgtcatcca
     1141 tcagagcgta tatgagctca tccatacaga agaccgggcg gaattccagc gccagcttca
     1201 ctgggctcta aacccagact ctgcacaagg agtggacgaa gcccatggcc ctccacaggc
     1261 agcagtctat tataccccag accagcttcc tccagagaac gcttctttca tggagaggtg
     1321 cttcaggtgc cggctgaggt gcctgctgga taattcatct ggttttctgg caatgaattt
     1381 ccaagggagg ttaaagtatc ttcatggaca gaacaagaaa gggaaggacg gagcgctgct
     1441 tcctccacaa ctggctttgt ttgcaatagc tactccactt cagccaccct ccatcctgga
     1501 aattcgaacc aaaaacttca tcttcaggac caaacacaag ctagacttca cacctattgg
     1561 ttgtgatgcc aaagggcagc ttattctggg ctatacagaa gtagagctgt gcacaagagg
     1621 atcggggtac cagttcatcc atgctgcaga catacttcac tgtgcagaat cccacatccg
     1681 catgattaag actggagaaa gtggcatgac agttttccgg cttcttgcaa aacacagtcg
     1741 ctggaggtgg gtccagtcca atgcacgctt gatttacaga aatggaagac cagattacat
     1801 catcgccact cagagaccac tgacggatga agaaggacga gagcatttac agaagcgaag
     1861 tacgtcgctg cccttcatgt ttgctaccgg agaggctgtg ttgtacgaga tctccagccc
     1921 tttctctccc ataatggatc ccctaccaat acgcaccaaa agcaacacta gcaggaaaga
     1981 ctgggctccc cagtcaaccc caagtaagga ttctttccac cccagttctc ttatgagtgc
     2041 cctcatccag caggatgagt ccatctatct gtgtcctcct tcaagccctg cgctgttaga
     2101 cagccatttt ctcatgggct ccgtgagcaa gtgcgggagt tggcaagaca gctttgcggc
     2161 cgcaggaagt gaggctgcgc tgaaacatga gcaaattggc catgctcagg acgtgaacct
     2221 tgcactctct ggcggcccct cagagctctt tccggataat aaaaataatg acttgtacag
     2281 catcatgagg aaccttggga ttgattttga agatatcaga agcatgcaga acgaggagtt
     2341 cttcagaact gactccaccg ctgctggtga ggttgacttc aaagacatcg acataacgga
     2401 cgaaatcctg acctacgtgc aggattccct gaacaattca actttgctga actcggcttg
     2461 ccagcagcag cctgtgactc agcacctaag ctgtatgctg caggagcgcc tgcaactaga
     2521 gcaacagcaa cagcttcagc agcccccgcc gcaggctctg gagccccagc agcagctgtg
     2581 tcagatggtg tgcccccagc aagatctggg tccgaagcac acgcaaatca acggcacgtt
     2641 tgcaagttgg aaccccaccc ctcccgtgtc tttcaactgt ccccagcagg aactaaagca
     2701 ctatcagctc ttttccagct tacaggggac tgctcaggaa tttccctaca aaccagaggt
     2761 ggacagtgtg ccttacacac agaactttgc tccctgtaat cagcctctgc ttccagaaca
     2821 ttccaagagt gtgcagttgg acttccctgg aagggatttt gaaccgtccc tgcatcccac
     2881 tacttctaat ttagattttg tcagttgttt acaagttcct gaaaaccaaa gtcatgggat
     2941 aaactcacag tccgccatgg tcagtcctca ggcatactat gctggggcca tgtccatgta
     3001 tcagtgccag ccagggccac agcgcacccc tgtggaccag acgcagtaca gctctgaaat
     3061 tccaggttct caggcattcc taagcaaggt gcagagtctc gaggaacaaa aactaatatc
     3121 agaggaagat ttggaacaga aactaatcag tgaagaagat ttagagcaaa agttaatttc
     3181 cgaggaggac ttatagtgag ggcccgatat ctcgacaatc aacctctgga ttacaaaatt
     3241 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     3301 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     3361 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     3421 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     3481 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     3541 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     3601 ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac ctggattctg
     3661 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     3721 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     3781 atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa acacggaagg
     3841 agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg
     3901 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     3961 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     4021 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     4081 atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     4141 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     4201 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4261 caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4321 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4381 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4441 acaacactca accctatctc ggtctattct tttgatttat aagggatttt ggggatttcg
     4501 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     4561 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4621 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     4681 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     4741 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     4801 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     4861 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     4921 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4981 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     5041 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     5101 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     5161 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     5221 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     5281 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     5341 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5401 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5461 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     5521 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5581 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5641 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5701 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5761 tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     5821 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     5881 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     5941 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     6001 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     6061 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     6121 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     6181 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     6241 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     6301 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     6361 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     6421 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     6481 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6541 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6601 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6661 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt
     6721 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6781 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6841 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6901 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     6961 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     7021 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     7081 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     7141 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     7201 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     7261 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     7321 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     7381 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     7441 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     7501 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     7561 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7621 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7681 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7741 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7801 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7861 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7921 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7981 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     8041 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     8101 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     8161 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     8221 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     8281 ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     8341 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     8401 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g
//
