LOCUS       V034608                 7655 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034608
VERSION     V034608
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             893..919
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             932..3175
                     /label="Smurf2(NM_025481)"
                     /note="Smurf2(NM_025481)"
                     /gene="Smurf2"
     polyA_signal    3222..3446
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3492..3920
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3934..4263
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4330..5124
                     /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    5298..5419
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5468..5484)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5492..5508
                     /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(5516..5546)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5561..5582
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5870..6455)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6626..7486)
                     /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(7487..7591)
                     /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 cttgccacca tgtaccctta
      901 tgacgtcccc gattacgccg gtaccgccac catgtctaac cccggaggcc ggaggaacgg
      961 gcccgtcaag ctgcgcctga cagtactctg tgcaaaaaac ttggtgaaaa aggatttttt
     1021 ccgtcttcct gatccatttg ctaaggtggt agttgatggt tctgggcagt gccattctac
     1081 agatactgtg aagaacacac ttgatccaaa gtggaatcag cattatgacc tgtatatcgg
     1141 aaagtctgat tcagttacga tcagcgtatg gaatcataag aagatccata agaagcaagg
     1201 tgctggattt cttggttgtg ttcgtcttct ttccaatgcc atcaaccgcc tcaaggacac
     1261 tggttatcag aggttggatt tatgcaaact ggggccaaat gacaatgata cagttcgagg
     1321 acaaattgta gtaagtcttc agtccagaga ccgaataggc acgggaggac aagttgtgga
     1381 ctgtagtcgt ttgtttgaca atgatttgcc agatggttgg gaagaaaggc ggactgcctc
     1441 gggaagaatc cagtatctaa accacatcac aagaactaca cagtgggaac gcccaacacg
     1501 accggcgtca gaatattcta gtcctggcag acccctcagc tgctttgtgg atgagaatac
     1561 tccaattact ggaacaaatg gtgcaacgtg tggacattct tcagacccca gactagcaga
     1621 gagaagagtc aggtcccagc gacatagaaa ttacatgagc aggacacact tacacactcc
     1681 tccagaccta ccggaaggct atgaacaaag gacaacgcag cagggtcagg tatatttctt
     1741 acatactcag actggtgtga gcacatggca tgatccacga gtgcctaggg atcttagcaa
     1801 catcaattgt gaagagctcg gtcctttgcc tcctggatgg gagatccgca ataccgcaac
     1861 aggaagagtt tatttcgttg accataacaa cagaacaaca caatttacag atccccggct
     1921 ctctgctaac ttgcatttag ttttaaatcg tcagaaccag ttgaaagacc aacagcaaca
     1981 gcaagtggtg ccattgtgtc ccgatgacac tgagtgtctg acagtgccaa gatacaagcg
     2041 agatttggtt caaaaactaa aaatcttgcg gcaagaactt tcccaacagc agcctcaagc
     2101 tggccactgc cgtattgagg tgtctaggga agagattttt gaggaatcat atagacaggt
     2161 catgaaaatg aggccaaaag atctatggaa gcgattaatg ataaaatttc gtggagaaga
     2221 aggtcttgac tatggaggag ttgccaggga atggttgtat ctcctgtcac atgaaatgtt
     2281 gaatccttac tatggtctct tccagtattc tagagatgat atctacacat tgcagatcaa
     2341 tcctgattct gcagttaatc cggaacattt atcctatttc cactttgttg gacggataat
     2401 gggaatggct gtattccatg gacattatat tgacggtggc ttcacattac ctttttataa
     2461 acagttgctt gggaagtcaa ttactttgga tgacatggag ctagttgacc cagaccttca
     2521 taatagttta gtgtggatac ttgaaaacga tatcacaggt gttctggacc ataccttctg
     2581 tgttgagcat aatgcatatg gtgaaataat ccagcatgaa ctcaaaccaa atggcaaaag
     2641 tatccctgtt actgaagaaa ataaaaaaga atatgtcagg ctttatgtca actggagatt
     2701 tctccgaggc atcgaagctc agttcctggc gctgcagaag ggctttaatg aagtcattcc
     2761 ccagcacctg ctgaagacat ttgatgagaa ggagctggag ctcattattt gtggacttgg
     2821 caaaattgat gtgagtgact ggaaggtcaa cacccggtta aaacactgta ccccagacag
     2881 caatgtcgtc aagtggttct ggaaagcggt ggagtttttt gatgaagaga gacgagcacg
     2941 gttgctccag tttgtgacag ggtcctctcg agtgcctctg cagggcttca aagcactaca
     3001 aggtgctgca ggcccacggc tctttaccat acaccagatt gatgcctgca cgaacaactt
     3061 gccaaaagcc cacacttgct tcaatcgaat agacattcca ccctatgaaa gctatgagaa
     3121 gctctatgaa aagctgctaa cggccatcga ggagacatgt ggctttgctg tggaatagct
     3181 cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     3241 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     3301 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     3361 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     3421 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     3481 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3541 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3601 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3661 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3721 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3781 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3841 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     3901 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3961 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4021 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4081 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4141 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4201 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4261 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     4321 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4381 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4441 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4501 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4561 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4621 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4681 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4741 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4801 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4861 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4921 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4981 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5041 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5101 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     5161 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5221 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5281 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5341 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5401 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     5461 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5521 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5581 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5641 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5701 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5761 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5821 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5881 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5941 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6001 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6061 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6121 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6181 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6241 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6301 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6361 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca
     6421 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6481 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6541 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6601 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6661 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6721 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6781 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6841 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6901 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6961 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7021 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7081 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7141 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7201 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7261 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7321 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7381 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7441 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7501 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7561 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7621 tcgacggatc gggagatctc ccgatcccct atggt
//
