LOCUS       V039346                 8290 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039346
VERSION     V039346
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..2949
                     /label="SMURF2(NM_022739)"
                     /note="SMURF2(NM_022739)"
                     /gene="SMURF2"
     CDS             2956..2982
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2986..3012
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3016..3042
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    3066..3654
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3720..3768
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3970..4398
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4412..4741
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4808..5602
                     /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    5778..5899
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5948..5964)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5972..5988
                     /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(5996..6026)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6041..6062
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6350..6938)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7109..7969)
                     /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(7970..8074)
                     /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 ccaccatgtc taaccccgga
      721 ggccggagga acgggcccgt caagctgcgc ctgacagtac tctgtgcaaa aaacctggtg
      781 aaaaaggatt ttttccgact tcctgatcca tttgctaagg tggtggttga tggatctggg
      841 caatgccatt ctacagatac tgtgaagaat acgcttgatc caaagtggaa tcagcattat
      901 gacctgtata ttggaaagtc tgattcagtt acgatcagtg tatggaatca caagaagatc
      961 cataagaaac aaggtgctgg atttctcggt tgtgttcgtc ttctttccaa tgccatcaac
     1021 cgcctcaaag acactggtta tcagaggttg gatttatgca aactcgggcc aaatgacaat
     1081 gatacagtta gaggacagat agtagtaagt cttcagtcca gagaccgaat aggcacagga
     1141 ggacaagttg tggactgcag tcgtttattt gataacgatt taccagacgg ctgggaagaa
     1201 aggagaaccg cctctggaag aatccagtat ctaaaccata taacaagaac tacgcaatgg
     1261 gagcgcccaa cacgaccggc atccgaatat tctagccctg gcagacctct tagctgcttt
     1321 gttgatgaga acactccaat tagtggaaca aatggtgcaa catgtggaca gtcttcagat
     1381 cccaggctgg cagagaggag agtcaggtca caacgacata gaaattacat gagcagaaca
     1441 catttacata ctcctccaga cctaccagaa ggctatgaac agaggacaac gcaacaaggc
     1501 caggtgtatt tcttacatac acagactggt gtgagcacat ggcatgatcc aagagtgccc
     1561 agggatctta gcaacatcaa ttgtgaagag cttggtccat tgcctcctgg atgggagatc
     1621 cgtaatacgg caacaggcag agtttatttc gttgaccata acaacagaac aacacaattt
     1681 acagatcctc ggctgtctgc taacttgcat ttagttttaa atcggcagaa ccaattgaaa
     1741 gaccaacagc aacagcaagt ggtatcgtta tgtcctgatg acacagaatg cctgacagtc
     1801 ccaaggtaca agcgagacct ggttcagaaa ctaaaaattt tgcggcaaga actttcccaa
     1861 caacagcctc aggcaggtca ttgccgcatt gaggtttcca gggaagagat ttttgaggaa
     1921 tcatatcgac aggtcatgaa aatgagacca aaagatctct ggaagcgatt aatgataaaa
     1981 tttcgtggag aagaaggcct tgactatgga ggcgttgcca gggaatggtt gtatctcttg
     2041 tcacatgaaa tgttgaatcc atactatggc ctcttccagt attcaagaga tgatatttat
     2101 acattgcaga tcaatcctga ttctgcagtt aatccggaac atttatccta tttccacttt
     2161 gttggacgaa taatgggaat ggctgtgttt catggacatt atattgatgg tggtttcaca
     2221 ttgccttttt ataagcaatt gcttgggaag tcaattacct tggatgacat ggagttagta
     2281 gatccggatc ttcacaacag tttagtgtgg atacttgaga atgatattac aggtgttttg
     2341 gaccatacct tctgtgttga acataatgca tatggtgaaa ttattcagca tgaacttaaa
     2401 ccaaatggca aaagtatccc tgttaatgaa gaaaataaaa aagaatatgt caggctctat
     2461 gtgaactgga gatttttacg aggcattgag gctcaattct tggctctgca gaaaggattt
     2521 aatgaagtaa ttccacaaca tctgctgaag acatttgatg agaaggagtt agagctcatt
     2581 atttgtggac ttggaaagat agatgttaat gactggaagg taaacacccg gttaaaacac
     2641 tgtacaccag acagcaacat tgtcaaatgg ttctggaaag ctgtggagtt ttttgatgaa
     2701 gagcgacgag caagattgct tcagtttgtg acaggatcct ctcgagtgcc tctgcagggc
     2761 ttcaaagcat tgcaaggtgc tgcaggcccg agactcttta ccatacacca gattgatgcc
     2821 tgcactaaca acctgccgaa agcccacact tgcttcaatc gaatagacat tccaccctat
     2881 gaaagctatg aaaagctata tgaaaagctg ctaacagcca ttgaagaaac atgtggattt
     2941 gctgtggaac tcgagtaccc atacgacgta cctgattatg caacttatcc ttatgatgta
     3001 cctgattatg ctagctatcc ttatgatgtc ccagactacg catagtgagg gcccgatatc
     3061 tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     3121 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     3181 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     3241 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     3301 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     3361 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     3421 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
     3481 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     3541 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     3601 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa
     3661 acgggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac
     3721 ggcaataaaa agacagaata aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg
     3781 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     3841 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     3901 cagccaacgt cggggcggca ggccctgcca tagcagatct gcgcagctgg ggctctaggg
     3961 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4021 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     4081 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt
     4141 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     4201 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4261 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     4321 ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac
     4381 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     4441 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     4501 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4561 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4621 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4681 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4741 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     4801 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4861 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4921 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4981 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     5041 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     5101 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     5161 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     5221 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     5281 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     5341 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     5401 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     5461 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     5521 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     5581 ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga
     5641 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5701 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5761 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5821 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5881 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     5941 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     6001 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     6061 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     6121 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     6181 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     6241 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6301 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6361 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6421 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6481 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6541 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6601 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6661 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6721 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6781 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6841 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6901 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6961 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     7021 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     7081 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     7141 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     7201 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     7261 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7321 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7381 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7441 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7501 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7561 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7621 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7681 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7741 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7801 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7861 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7921 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7981 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     8041 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     8101 acgtcgacgg atcgggagat ctcccgatcc cctatggtcg actctcagta caatctgctc
     8161 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
     8221 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
     8281 tctgcttagg
//
