LOCUS       V019502                 9332 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019502
VERSION     V019502
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..3988
                     /label="USP7(NM_003470)"
                     /note="USP7(NM_003470)"
                     /gene="USP7"
     CDS             3995..4084
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    4108..4696
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4762..4810
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5012..5440
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5454..5783
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5850..6644
                     /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    6820..6941
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6990..7006)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7014..7030
                     /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(7038..7068)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7083..7104
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7392..7980)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8151..9011)
                     /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(9012..9116)
                     /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 ccatgaacca ccagcagcag cagcagcagc agaaagcggg
      721 cgagcagcag ttgagcgagc ccgaggacat ggagatggaa gcgggagata cagatgaccc
      781 accaagaatt actcagaacc ctgtgatcaa tgggaatgtg gccctgagtg atggacacaa
      841 caccgcggag gaggacatgg aggatgacac cagttggcgc tccgaggcaa cctttcagtt
      901 cactgtggag cgcttcagca gactgagtga gtcggtcctt agccctccgt gttttgtgcg
      961 aaatctgcca tggaagatta tggtgatgcc acgcttttat ccagacagac cacaccaaaa
     1021 aagcgtagga ttctttctcc agtgcaatgc tgaatctgat tccacgtcat ggtcttgcca
     1081 tgcacaagca gtgctgaaga taataaatta cagagatgat gaaaagtcgt tcagtcgtcg
     1141 tattagtcat ttgttcttcc ataaagaaaa tgattgggga ttttccaatt ttatggcctg
     1201 gagtgaagtg accgatcctg agaaaggatt tatagatgat gacaaagtta cctttgaagt
     1261 ctttgtacag gcggatgctc cccatggagt tgcgtgggat tcaaagaagc acacaggcta
     1321 cgtcggctta aagaatcagg gagcgacttg ttacatgaac agcctgctac agacgttatt
     1381 tttcacgaat cagctacgaa aggctgtgta catgatgcca accgaggggg atgattcgtc
     1441 taaaagcgtc cctttagcat tacaaagagt gttctatgaa ttacagcata gtgataaacc
     1501 tgtaggaaca aaaaagttaa caaagtcatt tgggtgggaa actttagata gcttcatgca
     1561 acatgatgtt caggagcttt gtcgagtgtt gctcgataat gtggaaaata agatgaaagg
     1621 cacctgtgta gagggcacca tacccaaatt attccgcggc aaaatggtgt cctatatcca
     1681 gtgtaaagaa gtagactatc ggtctgatag aagagaagat tattatgata tccagctaag
     1741 tatcaaagga aagaaaaata tatttgaatc atttgtggat tatgtggcag tagaacagct
     1801 cgatggggac aataaatacg acgctgggga acatggctta caggaagcag agaaaggtgt
     1861 gaaattccta acattgccac cagtgttaca tctacaactg atgagattta tgtatgaccc
     1921 tcagacggac caaaatatca agatcaatga taggtttgaa ttcccagagc agttaccact
     1981 tgatgaattt ttgcaaaaaa cagatcctaa ggaccctgca aattatattc ttcatgcagt
     2041 cctggttcat agtggagata atcatggtgg acattatgtg gtttatctaa accccaaagg
     2101 ggatggcaaa tggtgtaaat ttgatgacga cgtggtgtca aggtgtacta aagaggaagc
     2161 aattgagcac aattatgggg gtcacgatga cgacctgtct gttcgacact gcactaatgc
     2221 ttacatgtta gtctacatca gggaatcaaa actgagtgaa gttttacagg cggtcaccga
     2281 ccatgatatt cctcagcagt tggtggagcg attacaagaa gagaaaagga tcgaggctca
     2341 gaagcggaag gagcggcagg aagcccatct ctatatgcaa gtgcagatag tcgcagagga
     2401 ccagttttgt ggccaccaag ggaatgacat gtacgatgaa gaaaaagtga aatacactgt
     2461 gttcaaagta ttgaagaact cctcgcttgc tgagtttgtt cagagcctct ctcagaccat
     2521 gggatttcca caagatcaaa ttcgattgtg gcccatgcaa gcaaggagta atggaacaaa
     2581 acgaccagca atgttagata atgaagccga cggcaataaa acaatgattg agctcagtga
     2641 taatgaaaac ccttggacaa tattcctgga aacagttgat cccgagctgg ctgctagtgg
     2701 agcgacctta cccaagtttg ataaagatca tgatgtaatg ttatttttga agatgtatga
     2761 tcccaaaacg cggagcttga attactgtgg gcatatctac acaccaatat cctgtaaaat
     2821 acgtgacttg ctcccagtta tgtgtgacag agcaggattt attcaagata ctagccttat
     2881 cctctatgag gaagttaaac cgaatttaac agagagaatt caggactatg acgtgtctct
     2941 tgataaagcc cttgatgaac taatggatgg tgacatcata gtatttcaga aggatgaccc
     3001 tgaaaatgat aacagtgaat tacccaccgc aaaggagtat ttccgagatc tctaccaccg
     3061 cgttgatgtc attttctgtg ataaaacaat ccctaatgat cctggatttg tggttacgtt
     3121 atcaaataga atgaattatt ttcaggttgc aaagacagtt gcacagaggc tcaacacaga
     3181 tccaatgttg ctgcagtttt tcaagtctca aggttatagg gatggcccag gtaatcctct
     3241 tagacataat tatgaaggta ctttaagaga tcttctacag ttcttcaagc ctagacaacc
     3301 taagaaactt tactatcagc agcttaagat gaaaatcaca gactttgaga acaggcgaag
     3361 ttttaaatgt atatggttaa acagccaatt tagggaagag gaaataacac tatatccaga
     3421 caagcatggg tgtgtccggg acctgttaga agaatgtaaa aaggccgtgg agcttgggga
     3481 gaaagcatca gggaaactta ggctgctaga aattgtaagc tacaaaatca ttggtgttca
     3541 tcaagaagat gaactattag aatgtttatc tcctgcaacg agccggacgt ttcgaataga
     3601 ggaaatccct ttggaccagg tggacataga caaagagaat gagatgcttg tcacagtggc
     3661 gcatttccac aaagaggtct tcggaacgtt cggaatcccg tttttgctga ggatacacca
     3721 gggcgagcat tttcgagaag tgatgaagcg aatccagagc ctgctggaca tccaggagaa
     3781 ggagtttgag aagtttaaat ttgcaattgt aatgatgggc cgacaccagt acataaatga
     3841 agacgagtat gaagtaaatt tgaaagactt tgagccacag cccggtaata tgtctcatcc
     3901 tcggccttgg ctagggctcg accacttcaa caaagcccca aagaggagtc gctacactta
     3961 ccttgaaaag gccattaaaa tccataacct cgaggaacaa aaactaatat cagaggaaga
     4021 tttggaacag aaactaatca gtgaagaaga tttagagcaa aagttaattt ccgaggagga
     4081 cttatagtga gggcccgata tctcgacaat caacctctgg attacaaaat ttgtgaaaga
     4141 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     4201 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     4261 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     4321 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     4381 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     4441 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     4501 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     4561 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     4621 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     4681 tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag gagacaatac
     4741 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac gggtgttggg
     4801 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     4861 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     4921 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcagat
     4981 ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     5041 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     5101 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     5161 aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     5221 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     5281 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     5341 aaccctatct cggtctattc ttttgattta taagggattt tggggatttc ggcctattgg
     5401 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     5461 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     5521 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5581 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     5641 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     5701 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     5761 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     5821 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     5881 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     5941 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     6001 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     6061 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     6121 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     6181 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     6241 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     6301 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     6361 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     6421 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     6481 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     6541 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     6601 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     6661 cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     6721 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     6781 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     6841 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     6901 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     6961 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     7021 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     7081 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     7141 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     7201 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     7261 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     7321 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     7381 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     7441 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     7501 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     7561 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     7621 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     7681 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     7741 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     7801 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     7861 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     7921 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7981 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     8041 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     8101 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     8161 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     8221 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     8281 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     8341 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     8401 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     8461 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     8521 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     8581 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     8641 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     8701 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     8761 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     8821 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     8881 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     8941 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     9001 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     9061 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     9121 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     9181 cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     9241 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     9301 tgaccgacaa ttgcatgaag aatctgctta gg
//
