LOCUS       V030331                 7308 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030331
VERSION     V030331
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..1988
                     /label="Zbp1(NM_021394)"
                     /note="Zbp1(NM_021394)"
                     /gene="Zbp1"
     CDS             1995..2060
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2084..2672
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2738..2786
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2988..3416
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3430..3759
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3826..4620
                     /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    4796..4917
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4966..4982)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4990..5006
                     /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(5014..5044)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5059..5080
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5368..5956)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6127..6987)
                     /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(6988..7092)
                     /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 agaagctcct gttgacttga
      781 gcacaggaga caatctggag caaaagatcc tgcaggtgtt gagcgatgac ggcggccctg
      841 tgaagattgg ccagctggtg aagaaatgcc aagtgcccaa gaaaaccctc aatcaagtcc
      901 tttaccgcct gaagaaggag gacagagtgt cctccccaga gcctgcaaca tggagcatag
      961 gcggggctgc ttctggagat ggggctcctg caatccctga gaactccagt gcccagccta
     1021 gccttgatga aagaatatta agattcctgg aagccaatgg gcctcacagg gccctgcaca
     1081 tcgccaaggc tctgggaatg acgacagcca aagaagtgaa cccactcctg tattccatga
     1141 gaaataagca ccttctgagc tatgacggac agacgtggaa gatctaccac tcacgtcagg
     1201 aaggccaaga catagctcat tctggagtca cacaagagtc ccctgcgatt atttgtcagc
     1261 acaatccggt caacatgatc tgccaacaag gagccaacag ccacatctcc attgccaatt
     1321 caaacgccat ccagattggt cacgggaatg tcatagtaag agagaaagcc tgtggtgagc
     1381 caggtcccag gacctctcac cctctaccct tggcatggga cgcctctgct caggacatgc
     1441 cccctgttgc ccacggagct cagtacatct acatggacaa gtccttgctc caacaagtgc
     1501 agcttggcca ccacaacgag atgagcctcg tgggggatgc agggaagcac ccctcttata
     1561 gcttttctga cagcccccca gaagtgtcaa ccaccactgc tgacccagga gcttcattca
     1621 acatgcaaac atctgagcca ggccctcacc ctgagggaga cacagtccag acagtccaca
     1681 tcaaatcctg ctttctagag gacgccacca ttggtaacgg caacaagatg accatccacc
     1741 ttaggtcaaa gggtgaagtc atggagtctg gagacagtga agagccgaag aaggaagaca
     1801 caggtacaag ctctgaagcc actccaccca gaagctgcca gcacacgccc agtgactcca
     1861 tgctgcccac ctctgagctg agagctatgg ctctgggaga cagtagcccc cagaccacag
     1921 aacctgtgct tagagagcat gaagtccagg acatagaaag ctctcaagac acaggactga
     1981 gcaagcaagg atccgactac aaagaccatg acggtgatta taaagatcat gacatcgact
     2041 acaaggatga cgatgacaag tagtgagggc ccgatatctc gacaatcaac ctctggatta
     2101 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     2161 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     2221 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     2281 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     2341 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     2401 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     2461 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     2521 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     2581 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     2641 gagtcggatc tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca
     2701 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     2761 acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     2821 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     2881 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     2941 ccctgccata gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag
     3001 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3061 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3121 tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca
     3181 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3241 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3301 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg
     3361 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3421 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3481 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3541 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3601 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3661 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     3721 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     3781 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3841 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3901 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3961 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4021 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4081 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4141 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4201 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4261 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4321 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4381 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4441 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4501 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4561 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4621 gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     4681 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4741 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     4801 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     4861 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     4921 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     4981 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5041 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5101 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5161 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5221 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5281 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5341 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5401 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5461 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5521 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     5581 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5641 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5701 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5761 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     5821 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5881 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5941 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6001 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6061 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6121 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6181 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6241 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6301 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6361 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6421 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6481 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6541 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6601 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6661 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6721 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6781 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6841 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6901 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6961 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7021 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7081 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     7141 cccgatcccc tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag
     7201 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     7261 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
