LOCUS       V034249                 7510 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034249
VERSION     V034249
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             706..2166
                     /label="TRIM50(NM_178125)"
                     /note="TRIM50(NM_178125)"
                     /gene="TRIM50"
     CDS             2173..2262
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    2286..2874
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2940..2988
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3190..3618
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3632..3961
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4028..4822
                     /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    4998..5119
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5168..5184)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5192..5208
                     /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(5216..5246)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5261..5282
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5570..6158)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6329..7189)
                     /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(7190..7294)
                     /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 ggatccacta gtccagtgtg gtggaattcg ccaccatggc ttggcaggtg
      721 agcctgccag agctggagga ccggcttcag tgtcccatct gcctggaggt cttcaaggag
      781 cccctgatgc tgcagtgtgg ccactcttac tgcaagggct gcctggtttc cctgtcctgc
      841 cacctggatg ccgagctgcg ctgccccgtg tgccggcagg cggtggacgg cagcagctcc
      901 ctgcccaacg tctccctggc cagggtgatc gaagccctga ggctccctgg ggacccggag
      961 cccaaggtct gcgtgcacca ccggaacccg ctcagccttt tctgcgagaa ggaccaggag
     1021 ctcatctgtg gcctctgcgg tctgctgggc tcccaccaac accacccggt cacgcccgtc
     1081 tccaccgtct acagccgcat gaaggaggag ctcgcagccc tcatctctga gctgaagcag
     1141 gagcagaaaa aggtggatga gctcatcgcc aaactggtga acaaccggac ccgaatcgtc
     1201 aatgagtcgg atgtcttcag ctgggtgatc cgccgcgagt tccaggagct gcaccacctg
     1261 gtggatgagg agaaggcccg ctgcctggag gggatagggg gtcacacccg tggcctggtg
     1321 gcctccctgg acatgcagct ggagcaggcc cagggaaccc gggagcggct ggcccaagcc
     1381 gagtgtgtgc tggaacagtt cggcaatgag gaccaccaca agttcatccg gaagttccac
     1441 tccatggcct ccagagcaga gatgccgcag gcccggccct tagaaggcgc attcagcccc
     1501 atctccttca agccaggcct ccaccaggct gacatcaagc tgaccgtgtg gaaaaggctc
     1561 ttccggaaag ttttgccagc cccggagcct ctcaagttgg accctgccac tgcccaccca
     1621 ctcctggagc tctccaaggg caacacggtg gtgcagtgcg ggcttctggc ccagcggcga
     1681 gccagccagc ctgagcgctt cgactacagc acctgcgtcc tggccagccg cggcttctcc
     1741 tgcggccgcc actactggga ggtggtggtg ggcagcaaga gcgactggcg cctgggggtc
     1801 atcaagggca cagccagccg taagggcaag ctgaacaggt cccccgagca cggcgtgtgg
     1861 ctgatcggcc tgaaggaggg ccgggtgtac gaagcctttg cctgcccccg ggtacccctg
     1921 cccgtggccg gccaccccca ccgcatcggg ctctacctgc actatgagca gggcgaactc
     1981 accttcttcg atgccgaccg ccccgatgac ctgcggccgc tctacacctt ccaggccgac
     2041 ttccagggca agctctaccc catcctggac acctgctggc acgagagggg cagcaactcg
     2101 ctgcccatgg tgctgccccc gcccagcggg cctggccccc tcagccccga gcagcccacc
     2161 aagctgctcg aggaacaaaa actaatatca gaggaagatt tggaacagaa actaatcagt
     2221 gaagaagatt tagagcaaaa gttaatttcc gaggaggact tatagtgagg gcccgatatc
     2281 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     2341 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     2401 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     2461 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     2521 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     2581 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     2641 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
     2701 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     2761 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     2821 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa
     2881 acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     2941 ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg
     3001 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     3061 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     3121 cagccaacgt cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg
     3181 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3241 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3301 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt
     3361 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3421 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3481 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     3541 ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac
     3601 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3661 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3721 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3781 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3841 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3901 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3961 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     4021 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4081 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4141 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4201 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4261 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4321 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4381 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4441 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4501 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     4561 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4621 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4681 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4741 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4801 ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga
     4861 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4921 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4981 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5041 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5101 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     5161 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5221 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5281 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5341 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5401 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5461 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5521 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5581 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5641 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5701 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5761 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5821 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5881 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5941 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6001 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6061 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6121 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6181 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6241 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6301 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6361 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6421 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6481 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6541 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6601 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6661 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6721 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6781 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6841 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6901 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6961 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7021 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7081 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7141 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7201 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7261 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7321 acgtcgacgg atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc
     7381 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
     7441 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
     7501 tctgcttagg
//
