LOCUS       V043919                 7357 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043919
VERSION     V043919
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             925..3108
                     /label="Smurf1(NM_001109598)"
                     /note="Smurf1(NM_001109598)"
                     /gene="Smurf1"
     CDS             3121..3186
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3221..3445
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3491..3919
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3933..4262
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4274..4873
                     /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    5033..5154
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5203..5219)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5227..5243
                     /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(5251..5281)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5296..5317
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5605..6190)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6361..7221)
                     /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(7222..7326)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 aagctttcta gagaattcgc caccatgtcg aaccccggga cccgtaggaa cggctccagc
      961 atcaagatcc gtctgacagt attgtgtgcc aagaaccttg caaagaaaga cttcttcaga
     1021 ctccccgacc cctttgccaa gattgttgtg gacggctctg ggcagtgcca ctcaaccgac
     1081 actgtgaaaa acaccctgga cccaaagtgg aaccagcact atgacctgta cgttgggaaa
     1141 accgactcga taaccatcag tgtgtggaac cacaagaaga tccacaagaa gcagggggct
     1201 ggcttcctgg gctgcgtgcg gctgctctcc aatgccatca gcagattgaa agataccggc
     1261 taccagcgtt tggatctatg caaactaaat ccctcagata ctgatgcagt tcgtggccaa
     1321 atagtggtca gtttacagac ccgagacaga ataggtggtg gagggtcggt ggtggactgc
     1381 aggggactgc tggagaacga aggaacggta tatgaagact caggccctgg aaggccgctc
     1441 agctgcctca tggaggaacc tgccccgtat acagatggca ccggtgcagc ggcaggaggt
     1501 gggaactgca ggtttgtgga gtctccaagc caagatcaga gactcctggt acagcgactc
     1561 cgaaatcctg aggttcgagg atccctacag acaccgcaga atcgaccaca tggccaccag
     1621 tcaccggagc tgcctgaagg ctacgagcaa aggacaacag tgcagggaca agtttacttt
     1681 ttgcacacgc agactggagt cagcacatgg cacgacccca ggatccccag agaccttaac
     1741 agtgtgaact gcgatgaact tgggccactg cctccaggct gggaagtcag aagcacagtg
     1801 tcgggaagaa tctattttgt agatcacaat aataggacaa cccagtttac agacccacgg
     1861 cttcaccaca tcatgaatca ccagtgccaa ctcaaggagc ccagccagcc gctgcagttg
     1921 cccagtgagg gctccgtgga ggacgaggag cttcctgccc agagatatga gagagattta
     1981 gtccagaagc ttaaagttct cagacatgag ctctctcttc agcagcccca agctggtcat
     2041 tgtcgcatag aagtttccag agaagaaata tttgaggagt catatcgcca gatcatgaag
     2101 atgcgaccaa aagacctgaa gaagcgcctg atggtgaagt tccgagggga ggaaggcttg
     2161 gactatggtg gggtggctcg ggagtggctg tatttgttgt gccatgaaat gttgaatccg
     2221 tattatggac tcttccagta ttccacggac aatatttaca cactgcaaat caacccagat
     2281 tcttctatca accctgacca tctgtcttac ttccactttg tgggtcgcat catgggtctg
     2341 gctgtgttcc atggacacta cataaacggg ggcttcacag ttcctttcta caagcagctc
     2401 ctgggaaaac ccatccagct gtcggatctg gagtcagtgg acccagaact tcacaagagc
     2461 ttggtgtgga ttctagagaa tgacatcacg ccagtgctgg atcacacctt ctgcgtggag
     2521 cacaatgctt ttgggcggat ccttcagcat gaactgaaac ccaatggcag aaatgtgcct
     2581 gtcactgagg agaacaagaa ggagtatgtg cggttgtatg tgaactggag gtttatgaga
     2641 ggaatcgaag cccagttcct agcgcttcag aaagggttta acgaactcat tccccaacac
     2701 ttactgaagc cctttgacca gaaggaacta gagctgataa taggcggcct ggataagata
     2761 gacctgaatg actggaagtc caacactcgg ctgaagcact gtgtggcaga cagcaacatc
     2821 gtcaggtggt tctggcaggc ggtggagacc ttcgatgagg agaggagagc caggctcctg
     2881 cagtttgtga caggatccac acgagttcct ctccaaggct tcaaggctct gcaaggcgcg
     2941 gcagggcccc ggctcttcac catccacctg attgacgcca acaccgacaa cctgcccaag
     3001 gcccacacct gctttaatcg gattgacatc ccaccctatg agtcctatga gaagctctat
     3061 gagaagctgc tgacagcggt agaggagacc tgtggctttg ccgtggagct cgagaccggt
     3121 gactacaaag accatgacgg tgattataaa gatcatgaca tcgattacaa ggatgacgat
     3181 gacaagtgac cggtaagttt aaaccgctga tcagcctcga ctgtgccttc tagttgccag
     3241 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     3301 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     3361 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     3421 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     3481 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3541 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     3601 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     3661 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     3721 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3781 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     3841 tttgatttat aagggatttt ggccatttcg gcctattggt taaaaaatga gctgatttaa
     3901 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3961 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     4021 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4081 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4141 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4201 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4261 aaaagctccc gggatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     4321 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     4381 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     4441 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     4501 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     4561 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     4621 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     4681 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     4741 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     4801 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     4861 gcccggtgcc tgattcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     4921 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4981 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     5041 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5101 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     5161 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     5221 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5281 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5341 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5401 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5461 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5521 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5581 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5641 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5701 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5761 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5821 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5881 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5941 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6001 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6061 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6121 acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6181 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6241 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6301 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6361 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6421 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6481 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6541 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6601 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6661 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6721 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6781 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6841 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6901 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6961 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7021 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7081 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7141 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7201 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7261 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7321 tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
