LOCUS       V045194                 8050 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V045194
VERSION     V045194
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..2709
                     /label="Mta2(NM_011842)"
                     /note="Mta2(NM_011842)"
                     /gene="Mta2"
     CDS             2716..2742
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2746..2772
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2776..2802
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    2826..3414
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3480..3528
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3730..4158
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4172..4501
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4568..5362
                     /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    5538..5659
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5708..5724)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5732..5748
                     /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(5756..5786)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5801..5822
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6110..6698)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6869..7729)
                     /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(7730..7834)
                     /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 ggctaacatg
      721 taccgggtgg gagattacgt ctattttgag aactcctcca gcaatcctta cctggttaga
      781 cggattgagg agctcaataa gactgcaaat ggaaatgtgg aagcaaaggt tgtctgtctt
      841 ttccggcgaa gggatatttc tagtagcctc aacagcttgg ctgacagtaa tgctcgggag
      901 tttgaggagg aatcaaaaca gccaggggtc tccgagcagc agagacatca actgaagcac
      961 cgagagcttt ttctttctcg gcagtttgaa tcattaccag ccacccacat aagggggaag
     1021 tgcagcgtga ctctcttgaa cgagacagac atcttaaacc agtatctaga caaggaggac
     1081 tgcttttttt actcactggt gtttgatcct gtgcagaaga cacttctggc tgatcaaggg
     1141 gagatcagag ttggttgcaa attccaagct gagatcccag atcgtttggc agagggagaa
     1201 tcagataatc gaaaccaaca gaagatggag atgaaagtct gggacccaga caaccctctc
     1261 acagaccggc agattgatca gtttctcgtg gtggcccgag ctgtgggaac cttcgcgaga
     1321 gccctagact gcagcagctc aattcggcag ccaagcttgc acatgagtgc agctgctgcg
     1381 tcccgagata tcactctgtt ccatgccatg gataccttgc aaaggaatgg ctatgacttg
     1441 gccaaggcta tgtcaaccct agtaccacag ggaggcccgg tgctctgtcg ggatgagatg
     1501 gaggaatggt ctgcttctga agccatgctg tttgaagagg ctctggagaa gtacgggaag
     1561 gatttcaatg atatccgcca ggactttcta ccctggaagt cacttgcaag catagtccag
     1621 ttttattaca tgtggaaaac cacagaccgg tatattcagc agaaaagatt gaaagctgct
     1681 gaagcagaca gcaaactgaa acaggtctac atccccactt ataccaagcc aaaccctaac
     1741 cagatcattt ctgtgggttc aaaacctggc atgaatgggg ctggattcca gaagggcctg
     1801 acttgtgaga gctgccacac cacacagtct gcccagtggt atgcctgggg accccctaac
     1861 atgcagtgcc gactttgtgc ttcctgttgg atttattgga agaaatacgg gggactgaag
     1921 acccctaccc aacttgaagg ggctgctcgg ggcaccacag agccacactc aagaggtcat
     1981 ttatccagac ctgaagccca aagtctctcc ccctatacaa ccagtgctaa ccgggccaag
     2041 ttgctggcta agaacaggca aactttcttg ctccagacca caaagctaac tcgtctagcc
     2101 agacgaatgt gccgagatct attacagcca aggagggctg cccgaagacc ctatgcacct
     2161 atcaatgcca atgccatcaa ggcagagtgc tcgatccgac ttcctaaggc tgccaagacg
     2221 ccactgaaaa ttcaccctct ggtgcggctg cccttggcaa ctatcgtcaa agatctggtg
     2281 gctcaggcac ctctgaaacc aaaaacacct cggggtacca agacaccaat caacagaaac
     2341 cagctgaccc agaatcgagg cctggggggc attatggtga aacgttccta tgagactatg
     2401 gcaggggcag gggtcccctt ctctgccaat ggaaggcctc tggcttcagg gattcgttca
     2461 agctcacagc cagcagccaa gcgtcagaaa ctaaaccctg ctgatgctcc caatccagtg
     2521 gtatttgtgg ccacaaagga caccagggct ttaaggaaag ctctaactca cctggaaatg
     2581 cgccgagcgg cccgaaggcc caacctaccc ctgaaggtga aaccaacgct gatgacagtg
     2641 cggcccccag ttcctctgcc cgcttcttca catcctgcca gcaccaatga gcccattgtc
     2701 ctggaggatc tcgagtaccc atacgacgta cctgattatg caacttatcc ttatgatgta
     2761 cctgattatg ctagctatcc ttatgatgtc ccagactacg catagtgagg gcccgatatc
     2821 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     2881 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     2941 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     3001 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     3061 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     3121 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     3181 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
     3241 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     3301 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     3361 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa
     3421 acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     3481 ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg
     3541 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     3601 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     3661 cagccaacgt cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg
     3721 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3781 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3841 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt
     3901 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3961 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4021 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     4081 ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac
     4141 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     4201 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     4261 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4321 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4381 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4441 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4501 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     4561 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4621 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4681 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4741 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4801 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4861 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4921 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4981 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     5041 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     5101 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     5161 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     5221 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     5281 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     5341 ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga
     5401 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5461 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5521 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5581 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5641 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     5701 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5761 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5821 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5881 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5941 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     6001 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6061 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6121 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6181 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6241 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6301 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6361 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6421 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6481 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6541 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6601 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6661 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6721 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6781 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6841 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6901 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6961 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     7021 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7081 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7141 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7201 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7261 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7321 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7381 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7441 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7501 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7561 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7621 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7681 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7741 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7801 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7861 acgtcgacgg atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc
     7921 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
     7981 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
     8041 tctgcttagg
//
