LOCUS       V043615                 7495 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043615
VERSION     V043615
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             931..3123
                     /label="Smurf1(NM_001038627)"
                     /note="Smurf1(NM_001038627)"
                     /gene="Smurf1"
     CDS             3130..3156
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3160..3186
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3190..3216
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    3257..3481
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3527..3955
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3969..4298
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4365..4964
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    5138..5259
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5308..5324)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5332..5348
                     /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(5356..5386)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5401..5422
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5710..6295)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6466..7326)
                     /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(7327..7431)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cactagtcca gtgtggtgga attcgccacc atgtcgaacc ccgggacccg taggaacggc
      961 tccagcatca agatccgtct gacagtattg tgtgccaaga accttgcaaa gaaagacttc
     1021 ttcagactcc ccgacccctt tgccaagatt gttgtggacg gctctgggca gtgccactca
     1081 accgacactg tgaaaaacac cctggaccca aagtggaacc agcactatga cctgtatgtt
     1141 gggaaaacgg actcgataac catcagtgtg tggaaccaca agaagatcca caagaagcag
     1201 ggggctggct tcctgggctg tgtgcggctg ctctccaatg ccatcagcag attgaaagac
     1261 actggctacc agcgtttgga tctatgcaaa ctaaatccct cagatactga tgcagttcgt
     1321 ggccaaatag tggtcagttt acagacccga gacagaatag gcggtggagg gtcagtggtg
     1381 gactgcagag ggctgctgga gaacgaagga acagtgtatg aagactcagg ccctggaagg
     1441 ccgctcagct gcctcatgga ggaacctgcc ccatatacag atggtactgg tgcagcagca
     1501 ggaggcggga actgcaggtt tgtggagtct ccaagccaag atcagagact cctggtacag
     1561 cgactccgaa atcctgaggt tcgagggccc ttacagacac cccagaaccg accacatggc
     1621 caccagtcgc cagagctgcc tgaaggctat gagcaaagga caacagtgca gggacaagtt
     1681 tactttttgc acacgcagac tggagtcagt acatggcatg accccaggat ccccagagac
     1741 cttaacagtg tgaactgcga tgaacttggg ccactgcctc caggctggga agtccgaagc
     1801 acagtgtcgg gaagaatcta ttttgtagat cacaataata ggacaaccca gtttacagat
     1861 ccacggcttc accacatcat gaatcaccag tgccaactca aggagcccag ccagccgctg
     1921 cagctgccca gtgagggctc cgtggaggac gaggagcttc ctgcccagag atatgagagg
     1981 gacttagtcc agaagctcaa agttctcagg cacgagctct ctcttcagca gccccaggct
     2041 ggtcactgtc gaatagaagt ctccagagaa gagatatttg aggagtcgta tcgccagatc
     2101 atgaagatgc ggccaaaaga cctaaagaag cgcctgatgg tgaagttccg aggggaggaa
     2161 ggtttggact atggtggagt ggctcgggag tggctgtatt tgttgtgcca tgaaatgttg
     2221 aacccgtact atggactctt ccagtattcc acggacaata tttacacact gcagatcaac
     2281 ccagattctt ctatcaaccc tgaccatctg tcatacttcc actttgtggg tcgcatcatg
     2341 ggtctggctg tgttccacgg acactacata aatgggggtt tcacagttcc gttctacaag
     2401 cagctcctgg ggaagccaat ccagctgtcg gacctggagt ccgtggaccc agaactgcat
     2461 aagagcttgg tgtggattct agagaatgac atcacgcctg tgttggatca taccttctgc
     2521 gtggagcaca acgctttcgg gcggattctc cagcatgaac tgaaacccaa tggcagaaat
     2581 gtgcctgtca ctgaggagaa caagaaggaa tacgtccggc tgtatgtgaa ctggaggttt
     2641 atgagaggaa tcgaagccca gttcttagca cttcagaagg ggtttaacga actcattccc
     2701 caacacttgc tgaagccctt tgaccagaag gaactagagc tgataatagg tgggctggat
     2761 aagatagacc tgaacgactg gaagtccaac acccggctga aacactgtgt ggcagacagc
     2821 aacatcgtca ggtggttctg gcaggcggtg gagaccttcg atgaggagag gagagccaga
     2881 ctcctgcagt ttgtgacagg atccacaaga gttcctctcc aaggcttcaa ggctctgcaa
     2941 ggctctacag gcgcggcagg gccccggctg ttcaccattc acctgataga cgccaataca
     3001 gacaacctgc ccaaggccca tacctgcttt aatcggatcg acatcccacc ctatgagtcc
     3061 tatgagaagc tctatgagaa gctgctgaca gcggtggagg aaacctgtgg ctttgcagtg
     3121 gagctcgagt acccatacga cgtacctgat tatgcaactt atccttatga tgtacctgat
     3181 tatgctagct atccttatga tgtcccagac tacgcatagt ctagagggcc cgtttaaacc
     3241 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     3301 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     3361 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     3421 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     3481 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     3541 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3601 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3661 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3721 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3781 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3841 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3901 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     3961 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4021 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4081 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4141 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4201 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     4261 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     4321 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgacc gagtacaagc
     4381 ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg
     4441 cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg
     4501 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg
     4561 tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg
     4621 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc
     4681 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg
     4741 ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc
     4801 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc
     4861 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg
     4921 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgagcggga ctctggggtt
     4981 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5041 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5101 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     5161 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5221 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5281 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5341 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5401 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5461 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5521 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5581 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5641 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5701 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5761 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5821 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5881 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5941 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6001 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6061 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6121 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6181 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6241 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6301 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6361 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6421 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6481 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6541 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6601 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6661 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6721 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6781 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6841 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6901 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6961 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7021 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7081 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7141 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7201 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7261 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7321 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7381 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7441 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggt
//
