LOCUS       V018426                 7762 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V018426
VERSION     V018426
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"
     regulatory      913..922
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             928..1017
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     CDS             1039..3282
                     /label="SMURF2(NM_022739)"
                     /note="SMURF2(NM_022739)"
                     /gene="SMURF2"
     polyA_signal    3362..3586
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3632..4060
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4074..4403
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4470..5264
                     /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    5438..5559
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5608..5624)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5632..5648
                     /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(5656..5686)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5701..5722
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6010..6595)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6766..7626)
                     /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(7627..7731)
                     /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 gtttaaactt aagccaccat ggcggcggaa caaaaactaa tatcagagga agatttggaa
      961 cagaaactaa tcagtgaaga agatttagag caaaagttaa tttccgagga ggacttatca
     1021 gcggaaaagc ttggtaccat gtctaacccc ggaggccgga ggaacgggcc cgtcaagctg
     1081 cgcctgacag tactctgtgc aaaaaacctg gtgaaaaagg attttttccg acttcctgat
     1141 ccatttgcta aggtggtggt tgatggatct gggcaatgcc attctacaga tactgtgaag
     1201 aatacgcttg atccaaagtg gaatcagcat tatgacctgt atattggaaa gtctgattca
     1261 gttacgatca gtgtatggaa tcacaagaag atccataaga aacaaggtgc tggatttctc
     1321 ggttgtgttc gtcttctttc caatgccatc aaccgcctca aagacactgg ttatcagagg
     1381 ttggatttat gcaaactcgg gccaaatgac aatgatacag ttagaggaca gatagtagta
     1441 agtcttcagt ccagagaccg aataggcaca ggaggacaag ttgtggactg cagtcgttta
     1501 tttgataacg atttaccaga cggctgggaa gaaaggagaa ccgcctctgg aagaatccag
     1561 tatctaaacc atataacaag aactacgcaa tgggagcgcc caacacgacc ggcatccgaa
     1621 tattctagcc ctggcagacc tcttagctgc tttgttgatg agaacactcc aattagtgga
     1681 acaaatggtg caacatgtgg acagtcttca gatcccaggc tggcagagag gagagtcagg
     1741 tcacaacgac atagaaatta catgagcaga acacatttac atactcctcc agacctacca
     1801 gaaggctatg aacagaggac aacgcaacaa ggccaggtgt atttcttaca tacacagact
     1861 ggtgtgagca catggcatga tccaagagtg cccagggatc ttagcaacat caattgtgaa
     1921 gagcttggtc cattgcctcc tggatgggag atccgtaata cggcaacagg cagagtttat
     1981 ttcgttgacc ataacaacag aacaacacaa tttacagatc ctcggctgtc tgctaacttg
     2041 catttagttt taaatcggca gaaccaattg aaagaccaac agcaacagca agtggtatcg
     2101 ttatgtcctg atgacacaga atgcctgaca gtcccaaggt acaagcgaga cctggttcag
     2161 aaactaaaaa ttttgcggca agaactttcc caacaacagc ctcaggcagg tcattgccgc
     2221 attgaggttt ccagggaaga gatttttgag gaatcatatc gacaggtcat gaaaatgaga
     2281 ccaaaagatc tctggaagcg attaatgata aaatttcgtg gagaagaagg ccttgactat
     2341 ggaggcgttg ccagggaatg gttgtatctc ttgtcacatg aaatgttgaa tccatactat
     2401 ggcctcttcc agtattcaag agatgatatt tatacattgc agatcaatcc tgattctgca
     2461 gttaatccgg aacatttatc ctatttccac tttgttggac gaataatggg aatggctgtg
     2521 tttcatggac attatattga tggtggtttc acattgcctt tttataagca attgcttggg
     2581 aagtcaatta ccttggatga catggagtta gtagatccgg atcttcacaa cagtttagtg
     2641 tggatacttg agaatgatat tacaggtgtt ttggaccata ccttctgtgt tgaacataat
     2701 gcatatggtg aaattattca gcatgaactt aaaccaaatg gcaaaagtat ccctgttaat
     2761 gaagaaaata aaaaagaata tgtcaggctc tatgtgaact ggagattttt acgaggcatt
     2821 gaggctcaat tcttggctct gcagaaagga tttaatgaag taattccaca acatctgctg
     2881 aagacatttg atgagaagga gttagagctc attatttgtg gacttggaaa gatagatgtt
     2941 aatgactgga aggtaaacac ccggttaaaa cactgtacac cagacagcaa cattgtcaaa
     3001 tggttctgga aagctgtgga gttttttgat gaagagcgac gagcaagatt gcttcagttt
     3061 gtgacaggat cctctcgagt gcctctgcag ggcttcaaag cattgcaagg tgctgcaggc
     3121 ccgagactct ttaccataca ccagattgat gcctgcacta acaacctgcc gaaagcccac
     3181 acttgcttca atcgaataga cattccaccc tatgaaagct atgaaaagct atatgaaaag
     3241 ctgctaacag ccattgaaga aacatgtgga tttgctgtgg aatgagaatt ctgcagatat
     3301 ccagcacagt ggcggccgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg
     3361 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     3421 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     3481 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     3541 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     3601 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     3661 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3721 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3781 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3841 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3901 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3961 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4021 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     4081 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4141 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4201 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4261 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4321 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     4381 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     4441 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4501 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4561 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4621 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     4681 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     4741 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     4801 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     4861 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     4921 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     4981 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     5041 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     5101 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     5161 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     5221 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     5281 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5341 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5401 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     5461 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5521 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5581 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5641 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     5701 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     5761 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5821 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5881 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5941 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6001 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6061 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6121 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6181 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6241 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6301 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6361 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6421 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6481 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6541 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6601 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6661 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6721 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6781 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6841 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6901 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6961 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7021 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7081 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7141 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7201 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7261 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7321 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7381 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7441 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7501 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7561 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7621 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7681 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7741 ccgaaaagtg ccacctgacg tc
//
