LOCUS       V019767                 8915 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019767
VERSION     V019767
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             683..3571
                     /label="USP4(NM_003363)"
                     /note="USP4(NM_003363)"
                     /gene="USP4"
     CDS             3578..3667
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    3691..4279
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4345..4393
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4595..5023
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5037..5366
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5433..6227
                     /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    6403..6524
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6573..6589)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6597..6613
                     /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(6621..6651)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6666..6687
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6975..7563)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7734..8594)
                     /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(8595..8699)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatggcgga aggtggaggc tgccgtgagc gaccggatgc
      721 ggagactcag aagtccgagc ttggaccctt aatgaggacc acactccaac gcggggcgca
      781 gtggtatctt attgacagcc ggtggttcaa gcagtggaag aagtatgtgg gctttgacag
      841 ctgggacatg tacaatgtgg gtgaacataa cctatttcct ggcccaatag acaactctgg
      901 gctattttca gatcctgaga gtcagacctt gaaagaacac ttaattgatg aattggacta
      961 tgtattggtc cctaccgagg cgtggaataa actactaaac tggtacggct gtgtagaagg
     1021 ccagcaaccc atcgtcagaa aagttgtgga gcatggcctg tttgtcaagc actgcaaagt
     1081 cgaggtgtat ttgctggaac tgaagctctg tgagaacagt gaccccacca atgtgctgag
     1141 ttgccatttc agcaaggcag acaccattgc aaccatcgag aaagagatgc ggaagctatt
     1201 caacatccct gcggagcgtg aaacacggct ctggaacaaa tacatgagca acacctacga
     1261 gcagttgagc aagctagaca acactgtcca ggatgctggg ctataccagg gtcaggtgct
     1321 agtaattgag cctcaaaatg aagatggcac atggcccagg cagaccttgc agtcaaaatc
     1381 aagcactgcg cctagcagaa attttactac ctctccaaaa tcatcagcaa gtccctattc
     1441 ctcagtgtct gcctctctca ttgcaaatgg tgatagcact agcacctgtg ggatgcacag
     1501 ttccggtgtc agcaggggtg gatctggctt ttctgcttcg tataattgtc aggagccacc
     1561 atcctctcat atacaacctg ggctctgtgg acttggaaac ctgggaaaca cctgcttcat
     1621 gaactccgct ttgcagtgtt tgagcaacac tgcaccactg actgactact ttctcaaaga
     1681 tgagtatgaa gccgaaatca acagagacaa ccctctgggg atgaaagggg aaattgcaga
     1741 agcctatgct gaactcatta agcagatgtg gtctggaagg gacgcccatg tggcacctcg
     1801 catgttcaaa actcaagtag gacgttttgc tcctcaattt tctggctacc agcaacaaga
     1861 ttctcaggag ctgctggcct ttcttctaga tggattgcat gaagatctga accgggtaaa
     1921 gaaaaagccc tacttggagc tgaaggatgc caatgggcgg ccagatgcgg tggtggcaaa
     1981 ggaagcctgg gagaatcaca ggttgaggaa tgattctgtg attgtggata ctttccatgg
     2041 cctcttcaaa tctactttgg tttgcccaga atgtgctaag gtttctgtga cctttgaccc
     2101 attttgctat ctaacgctgc cactgccctt gaagaaagat cgagttatgg aggttttcct
     2161 ggttcctgct gaccctcact gcagacctac tcagtaccgt gtgactgtgc cgctgatggg
     2221 ggctgtgtcc gacctgtgcg aggctctctc caggctgtct ggcattgctg cagaaaatat
     2281 ggtggtcgca gatgtgtata atcaccgatt ccacaaaatt ttccaaatgg atgaaggttt
     2341 aaaccacatc atgcctcggg atgacatttt cgtgtacgag gtctgcagca cttccgtgga
     2401 tggctcggaa tgtgtcacgc ttccagtcta cttcagggag aggaagtcca ggccatcaag
     2461 cacttcctcc gcatcagcgc tatatgggca gccactattg ctttctgtcc ccaagcacaa
     2521 gttaaccctt gagtctttgt accaggctgt ttgtgatcgt atcagccgct atgtgaaaca
     2581 gcctttacct gatgagtttg gcagctcacc cttggagcca ggggcctgca atggctccag
     2641 gaacagctgt gaaggagaag atgaggaaga aatggagcat caggaagaag gcaaagagca
     2701 gctttcagaa acagaaggca gtggggaaga tgagccagga aatgacccca gtgagaccac
     2761 ccaaaagaag atcaaaggcc agccctgccc aaaaaggctt tttaccttca gtcttgtgaa
     2821 ctcctatgga acagctgaca taaattcact tgcagctgat ggaaaactac ttaaactcaa
     2881 ctctcgatct acactggcca tggattggga cagtgaaact cggagacttt actatgatga
     2941 gcaagaatct gaggcctacg agaagcatgt gagcatgttg cagcctcaga agaagaagaa
     3001 gaccacagtg gccctgagag actgcatcga gctcttcacc accatggaga cccttgggga
     3061 gcatgacccc tggtactgtc ccaactgtaa gaagcatcaa caggccacaa aaaagtttga
     3121 cctatggtcc ttgcccaaga tcctggtggt ccacctcaaa cgtttctcct acaacagata
     3181 ctggagggat aagctcgaca cagtcgtaga attcccaatc agagggctga acatgtccga
     3241 gtttgtctgt aacctgtcag caaggcctta tgtgtacgac ctcattgccg tgtccaatca
     3301 ttatggagcc atgggggttg gccactacac tgcatatgcg aagaacaaac tgaatggtaa
     3361 atggtattac tttgatgata gcaacgtgtc cctggcctct gaggatcaga tagtgactaa
     3421 agcagcttat gtgctatttt accaacgtcg agatgatgaa ttttataaga caccttcact
     3481 tagcagttct ggttcctctg atggagggac acgaccaagc agctctcagc agggctttgg
     3541 ggatgatgag gcttgcagca tggacaccaa cctcgaggaa caaaaactaa tatcagagga
     3601 agatttggaa cagaaactaa tcagtgaaga agatttagag caaaagttaa tttccgagga
     3661 ggacttatag tgagggcccg atatctcgac aatcaacctc tggattacaa aatttgtgaa
     3721 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     3781 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     3841 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     3901 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     3961 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     4021 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     4081 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     4141 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     4201 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     4261 ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg aaggagacaa
     4321 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt
     4381 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     4441 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     4501 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagca
     4561 gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     4621 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4681 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4741 ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4801 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4861 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4921 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat
     4981 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     5041 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5101 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     5161 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     5221 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5281 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     5341 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     5401 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     5461 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     5521 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     5581 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     5641 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5701 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5761 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5821 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     5881 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5941 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     6001 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     6061 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     6121 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     6181 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     6241 gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     6301 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     6361 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     6421 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     6481 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     6541 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     6601 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     6661 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     6721 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     6781 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     6841 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     6901 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6961 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     7021 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     7081 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     7141 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     7201 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     7261 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     7321 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     7381 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     7441 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7501 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7561 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7621 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7681 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7741 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7801 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7861 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7921 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7981 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     8041 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     8101 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     8161 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     8221 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     8281 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     8341 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     8401 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     8461 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     8521 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     8581 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8641 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8701 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
     8761 ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
     8821 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
     8881 gcttgaccga caattgcatg aagaatctgc ttagg
//
