LOCUS       V045646                 7158 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045646
VERSION     V045646
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             756..1838
                     /label="Oas1d(NM_133893)"
                     /note="Oas1d(NM_133893)"
                     /gene="Oas1d"
     CDS             1845..1910
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1934..2522
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2588..2636
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2838..3266
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3280..3609
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3676..4470
                     /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    4646..4767
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4816..4832)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4840..4856
                     /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(4864..4894)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4909..4930
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5218..5806)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5977..6837)
                     /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(6838..6942)
                     /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 ccaccatggc gagggaactc ttcagaaccc
      781 caatctggag gctggataag ttcatagagg atcaactcct tcctgacacc accttcctta
      841 ctgagctcag agcagacatc gactccataa gtgctttcct gatggagaga tgcttccaag
      901 gtgccgccca tcctgtgagg gtctccaggg ttgtgatggg tggctgctac aatgaataca
      961 ctgtgctcaa gggcaggtct gaggccaaca tggtggtgtt ccttatcaat ctcacaagct
     1021 ttgaggatca gttcaacgga caggtagtgt tcattgagga aatttggaga cacctactcc
     1081 agttgcagca agagaaacta tgtaaactca agtttgaggt ccagagccca aaggagccca
     1141 actccaggtt tctgagcttc aagctgagct gccccgagcg ccagcatgag ttggaatttg
     1201 atgtgcagcc agcctatgat gccctgtatg aagtaagaca cttcaagccc tttgactcca
     1261 gtaactacaa caaagtctac gcccaactca cccatgagtg caccacactg gagaaggagg
     1321 gcgagttctc catctgcttc accgacctcc atcagagctt cctgaggtat cgtgcgccca
     1381 agctctggaa cctcatccgt ttggtcaagc actggtatca actgtgtaag gagaagctga
     1441 gggggccgct gcctccacag tacgccctgg agctgctcac agtctacgtc tgggaatacg
     1501 ggatccacga aaaccctgga ctccatacag cccagtgctt ccgcactgtc ttagaacttg
     1561 tcaccaagta caaacggctt cgaatctact ggacatggtg ttatgacttt caacacgaga
     1621 tctctgacta cctgcaggga cagatcaaaa aagccaggcc tctgatcctg gatccagcag
     1681 acccaacaag gaatgtggct ggttcagact tacaggcatg ggacctgctg gcaaaggagg
     1741 ctcagatctg gatagattcg actttcttta cgaaccatga tatgtccatt gtggaagcct
     1801 gggaagtgat gccagagaga caagaatgtg tcttcctggg atccgactac aaagaccatg
     1861 acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc
     1921 ccgatatctc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     1981 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     2041 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     2101 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     2161 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     2221 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     2281 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     2341 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     2401 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     2461 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     2521 gcctggaaac gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     2581 gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa
     2641 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     2701 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     2761 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg
     2821 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2881 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     2941 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc
     3001 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3061 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3121 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3181 ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct
     3241 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     3301 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3361 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3421 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3481 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3541 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3601 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     3661 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3721 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3781 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3841 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3901 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3961 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4021 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4081 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4141 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4201 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4261 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4321 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4381 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4441 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga
     4501 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     4561 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4621 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     4681 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4741 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     4801 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4861 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4921 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4981 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5041 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5101 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5161 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5221 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5281 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5341 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5401 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5461 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5521 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5581 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5641 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5701 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5761 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5821 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5881 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5941 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6001 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6061 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6121 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6181 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6241 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6301 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6361 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6421 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6481 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6541 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6601 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6661 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6721 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6781 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6841 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6901 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6961 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca
     7021 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     7081 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     7141 atgaagaatc tgcttagg
//
