LOCUS       V034250                 7218 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034250
VERSION     V034250
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..1898
                     /label="Ptx3(NM_008987)"
                     /note="Ptx3(NM_008987)"
                     /gene="Ptx3"
     CDS             1905..1970
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1994..2582
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2648..2696
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2898..3326
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3340..3669
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3736..4530
                     /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    4706..4827
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4876..4892)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4900..4916
                     /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(4924..4954)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4969..4990
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5278..5866)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6037..6897)
                     /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(6898..7002)
                     /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 ccaccatgca cctccctgcg atcctgcttt
      781 gtgctctctg gtctgcagta gtggctgaga cctcggatga ctacgagctc atgtatgtga
      841 atttggacaa cgaaatagac aatggacttc atcccaccga ggaccccacg ccatgcgact
      901 gccgccagga gcactcggag tgggacaagc tgttcatcat gctggagaac tcgcagatgc
      961 gggagggcat gctgttgcag gccaccgacg acgtcctccg tggagagctg cagcggctgc
     1021 gggcagagct ggggcggctg gcgggcggca tggcgaggcc gtgcgcagcc ggtggccccg
     1081 cagacgccag gctggtgcgg gcgctggagc cgctgctgca ggagagccgt gacgcgagcc
     1141 tcaggctggc gcgcctggag gacgcggagg cgcggcgacc cgaggcgaca gtgcctggcc
     1201 taggcgctgt gctggaggaa ctgcggcgga cgcgcgccga cctgagcgcc gtgcagagct
     1261 gggtcgcccg ccactggctg cccgcaggtt gtgaaacagc aattttcttc ccaatgcgtt
     1321 cgaagaagat ttttggaagc gtgcatcctg tgagaccaat gaagcttgaa tcttttagta
     1381 cttgcatttg ggtcaaagcc acagatgtat taaacaaaac catcctgttt tcttatggca
     1441 caaagtggaa cccctatgag attcagctgt acctcagttc ccagtcccta gtgttggtgg
     1501 tgggtggaaa ggagaacaag ctggctgcag acactgtggt gtccctgggg aggtggtccc
     1561 acctgtgtgg cacctggagt tcagagcagg ggagcatgtc cctgtgggca aacggggagc
     1621 tggtggctac cactgtagag atggccaaaa gtcactctgt tcctgagggt ggactcctac
     1681 agattggcca agaaaagaat ggttgctgtg taggtggggg ctttgacgaa tcattagcat
     1741 tttctggaag aatcacaggc ttcaatatct gggatcgggt tctcagcgag gaggagatac
     1801 gggccagtgg aggagtcgaa tcctgtcaca tccggggaaa tgtcgtcggg tggggagtca
     1861 cagagattca ggcgcacgga ggagcccagt atgtttctgg atccgactac aaagaccatg
     1921 acggtgatta taaagatcat gacatcgact acaaggatga cgatgacaag tagtgagggc
     1981 ccgatatctc gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     2041 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     2101 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     2161 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     2221 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     2281 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     2341 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     2401 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     2461 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     2521 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     2581 gcctggaaac gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     2641 gctatgacgg caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa
     2701 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     2761 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     2821 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg
     2881 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2941 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3001 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc
     3061 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3121 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3181 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3241 ctattctttt gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct
     3301 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     3361 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3421 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3481 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3541 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3601 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3661 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     3721 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3781 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3841 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3901 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3961 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     4021 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     4081 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4141 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4201 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4261 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4321 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4381 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4441 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4501 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga
     4561 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     4621 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     4681 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     4741 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4801 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     4861 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4921 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4981 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5041 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5101 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5161 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     5221 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     5281 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     5341 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     5401 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     5461 ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     5521 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     5581 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5641 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5701 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5761 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5821 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5881 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5941 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6001 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6061 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6121 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6181 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     6241 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     6301 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     6361 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     6421 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     6481 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     6541 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6601 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6661 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6721 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6781 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6841 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6901 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6961 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7021 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca
     7081 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     7141 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     7201 atgaagaatc tgcttagg
//
