LOCUS       V019760                 7535 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019760
VERSION     V019760
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             908..2971
                     /label="USP49(NM_001286554)"
                     /note="USP49(NM_001286554)"
                     /gene="USP49"
     CDS             2978..3067
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     polyA_signal    3102..3326
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3372..3800
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3814..4143
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4210..5004
                     /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    5178..5299
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5348..5364)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5372..5388
                     /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(5396..5426)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5441..5462
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5750..6335)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6506..7366)
                     /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(7367..7471)
                     /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 cgccaccatg gatagatgca aacatgtagg gcggttacgg ctcgcccagg accactccat
      961 cctgaaccct cagaagtggt gctgcttaga gtgtgccacc accgagtccg tgtgggcctg
     1021 cctcaagtgc tcccacgtgg cctgcggccg ctatattgag gaccacgccc tgaaacactt
     1081 tgaggagacg ggacacccgc tagccatgga agtccgggat ctctacgtgt tctgttacct
     1141 gtgcaaggac tacgtgctca atgataaccc agagggggac ctgaagctgc taagaagctc
     1201 cctcctggcg gtccggggcc agaaacagga cacgccggtg agacgtgggc ggacgctgcg
     1261 gtccatggct tcgggtgagg acgtggtcct gccgcagcgc gctcctcagg gacagccgca
     1321 gatgctcacg gctctgtggt accggcgtca gcgcctgctg gccaggacgc tgcggctgtg
     1381 gttcgagaag agctcccggg gccaggcgaa gctggagcag cggcggcagg aggaggccct
     1441 ggagcgcaag aaggaggagg cgcggaggcg gcggcgcgag gtgaaacggc ggctgctgga
     1501 ggagctggcc agcacccctc cgcgcaagag tgcacggctg ctcctgcaca cgccccgcga
     1561 cgcgggcccg gctgcctcgc gccccgccgc cctccctacc tcacgcagag tgcccgccgc
     1621 cacactcaag ctgcgtcgcc agccggccat ggccccaggc gtcacgggcc tgcgcaacct
     1681 gggcaacacc tgctacatga actccatcct ccaggtgctc agccacctcc agaagttccg
     1741 agaatgtttc ctcaaccttg acccttccaa aacggaacat ctgtttccca aagccaccaa
     1801 cgggaagact cagctttctg gcaagccaac caacagctcg gccacggagc tgtccttgag
     1861 aaatgacagg gccgaggcat gcgagcggga gggcttctgc tggaacggca gggcctccat
     1921 tagtcggagt ctggagctca tccagaacaa ggagccgagt tcaaagcaca tttccctctg
     1981 ccgtgaactg cacaccctct tccgagtcat gtggtccggg aagtgggccc tagtgtcgcc
     2041 cttcgccatg ctccactcag tgtggagcct gatccctgcc ttccgcggct acgaccaaca
     2101 ggacgcgcag gaatttctct gcgagctgct gcacaaggtg cagcaggaac tcgagtctga
     2161 gggcaccaca cgccggatcc tcatcccctt ctcccagagg aagctcacca aacaggtctt
     2221 aaaggtggtg aataccatat ttcatgggca gctgctcagt caggtcacat gtatatcatg
     2281 caattacaaa tccaatacca ttgagccctt ttgggaccta tccctggaat tccctgaacg
     2341 ctatcactgc atagaaaagg ggtttgtccc tttgaatcaa acagagtgct tgctcactga
     2401 gatgctggcc aaattcacag agacagaggc cctggaaggg agaatctacg cttgtgacca
     2461 gtgtaacagc aaacgacgaa aatccaatcc caaacccctt gttctgagtg aagctagaaa
     2521 gcagttaatg atctacagac tacctcaggt tctccggctg caccttaaaa gattcaggtg
     2581 gtctggccgt aatcatcgag agaagattgg ggtccatgtc gtctttgacc aggtattaac
     2641 catggaacct tactgctgca gggacatgct ctcctctctt gacaaagaga cctttgccta
     2701 tgatctctcc gcagtggtca tgcatcacgg gaaagggttt ggctcaggac actacacagc
     2761 ctattgctac aacacagagg gaggtttttg ggtccactgc aatgactcaa agctgaatgt
     2821 atgcagtgtc gaggaagtgt gcaaaaccca ggcctacatc cttttttaca ctcaaagaac
     2881 agtgcagggc aatgcaagaa tctcagaaac ccatctccaa gctcaggtgc agtccagcaa
     2941 caatgatgaa ggcagaccac agacattttc cctcgaggaa caaaaactaa tatcagagga
     3001 agatttggaa cagaaactaa tcagtgaaga agatttagag caaaagttaa tttccgagga
     3061 ggacttatga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     3121 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     3181 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     3241 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     3301 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     3361 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3421 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3481 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3541 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3601 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3661 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3721 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     3781 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3841 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3901 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3961 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4021 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4081 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4141 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     4201 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4261 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4321 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4381 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4441 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4501 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4561 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4621 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4681 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4741 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4801 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4861 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4921 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4981 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     5041 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5101 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5161 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5221 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5281 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     5341 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5401 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5461 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5521 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5581 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5641 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5701 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5761 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5821 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5881 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5941 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6001 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6061 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6121 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6181 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6241 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca
     6301 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6361 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6421 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6481 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6541 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6601 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6661 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6721 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6781 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6841 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6901 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6961 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7021 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7081 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7141 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7201 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7261 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7321 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7381 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7441 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7501 tcgacggatc gggagatctc ccgatcccct atggt
//
