LOCUS       V027481                 7654 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027481
VERSION     V027481
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"
     regulatory      925..934
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             934..975
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /transl_table=1
                     /translation="GKPIPNPLLGLDST"
                     /label="V5 tag"
     CDS             982..3174
                     /label="Smurf1(NM_001038627)"
                     /note="Smurf1(NM_001038627)"
                     /gene="Smurf1"
     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             4329..5123
                     /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    5297..5418
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5467..5483)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5491..5507
                     /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(5515..5545)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5560..5581
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5869..6454)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6625..7485)
                     /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(7486..7590)
                     /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 atgggtaagc ctatccctaa ccctctcctc
      961 ggtctcgatt ctacagatat catgtcgaac cccgggaccc gtaggaacgg ctccagcatc
     1021 aagatccgtc tgacagtatt gtgtgccaag aaccttgcaa agaaagactt cttcagactc
     1081 cccgacccct ttgccaagat tgttgtggac ggctctgggc agtgccactc aaccgacact
     1141 gtgaaaaaca ccctggaccc aaagtggaac cagcactatg acctgtatgt tgggaaaacg
     1201 gactcgataa ccatcagtgt gtggaaccac aagaagatcc acaagaagca gggggctggc
     1261 ttcctgggct gtgtgcggct gctctccaat gccatcagca gattgaaaga cactggctac
     1321 cagcgtttgg atctatgcaa actaaatccc tcagatactg atgcagttcg tggccaaata
     1381 gtggtcagtt tacagacccg agacagaata ggcggtggag ggtcagtggt ggactgcaga
     1441 gggctgctgg agaacgaagg aacagtgtat gaagactcag gccctggaag gccgctcagc
     1501 tgcctcatgg aggaacctgc cccatataca gatggtactg gtgcagcagc aggaggcggg
     1561 aactgcaggt ttgtggagtc tccaagccaa gatcagagac tcctggtaca gcgactccga
     1621 aatcctgagg ttcgagggcc cttacagaca ccccagaacc gaccacatgg ccaccagtcg
     1681 ccagagctgc ctgaaggcta tgagcaaagg acaacagtgc agggacaagt ttactttttg
     1741 cacacgcaga ctggagtcag tacatggcat gaccccagga tccccagaga ccttaacagt
     1801 gtgaactgcg atgaacttgg gccactgcct ccaggctggg aagtccgaag cacagtgtcg
     1861 ggaagaatct attttgtaga tcacaataat aggacaaccc agtttacaga tccacggctt
     1921 caccacatca tgaatcacca gtgccaactc aaggagccca gccagccgct gcagctgccc
     1981 agtgagggct ccgtggagga cgaggagctt cctgcccaga gatatgagag ggacttagtc
     2041 cagaagctca aagttctcag gcacgagctc tctcttcagc agccccaggc tggtcactgt
     2101 cgaatagaag tctccagaga agagatattt gaggagtcgt atcgccagat catgaagatg
     2161 cggccaaaag acctaaagaa gcgcctgatg gtgaagttcc gaggggagga aggtttggac
     2221 tatggtggag tggctcggga gtggctgtat ttgttgtgcc atgaaatgtt gaacccgtac
     2281 tatggactct tccagtattc cacggacaat atttacacac tgcagatcaa cccagattct
     2341 tctatcaacc ctgaccatct gtcatacttc cactttgtgg gtcgcatcat gggtctggct
     2401 gtgttccacg gacactacat aaatgggggt ttcacagttc cgttctacaa gcagctcctg
     2461 gggaagccaa tccagctgtc ggacctggag tccgtggacc cagaactgca taagagcttg
     2521 gtgtggattc tagagaatga catcacgcct gtgttggatc ataccttctg cgtggagcac
     2581 aacgctttcg ggcggattct ccagcatgaa ctgaaaccca atggcagaaa tgtgcctgtc
     2641 actgaggaga acaagaagga atacgtccgg ctgtatgtga actggaggtt tatgagagga
     2701 atcgaagccc agttcttagc acttcagaag gggtttaacg aactcattcc ccaacacttg
     2761 ctgaagccct ttgaccagaa ggaactagag ctgataatag gtgggctgga taagatagac
     2821 ctgaacgact ggaagtccaa cacccggctg aaacactgtg tggcagacag caacatcgtc
     2881 aggtggttct ggcaggcggt ggagaccttc gatgaggaga ggagagccag actcctgcag
     2941 tttgtgacag gatccacaag agttcctctc caaggcttca aggctctgca aggctctaca
     3001 ggcgcggcag ggccccggct gttcaccatt cacctgatag acgccaatac agacaacctg
     3061 cccaaggccc atacctgctt taatcggatc gacatcccac cctatgagtc ctatgagaag
     3121 ctctatgaga agctgctgac agcggtggag gaaacctgtg gctttgcagt ggagtgactc
     3181 gagtctagag ggcccgttta aacccgctga 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 gccgatttcg 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 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     4321 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4381 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4441 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4501 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4561 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4621 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4681 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4741 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4801 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     4861 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4921 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4981 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     5041 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     5101 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     5161 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     5221 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5281 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5341 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5401 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     5461 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     5521 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5581 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5641 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5701 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5761 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5821 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5881 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5941 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6001 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     6061 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6121 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6181 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6241 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6301 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6361 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt ttgtttgcaa
     6421 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     6481 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6541 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6601 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6661 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6721 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6781 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6841 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6901 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6961 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7021 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     7081 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     7141 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7201 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7261 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7321 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7381 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7441 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7501 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7561 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     7621 cgacggatcg ggagatctcc cgatccccta tggt
//
