LOCUS       V031575                 9285 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031575
VERSION     V031575
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             706..4011
                     /label="USP7(NM_003470)"
                     /note="USP7(NM_003470)"
                     /gene="USP7"
     CDS             4018..4041
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHHHH"
                     /label="8xHis"
     misc_feature    4061..4649
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4715..4763
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4965..5393
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5407..5736
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5803..6597
                     /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    6773..6894
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6943..6959)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6967..6983
                     /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(6991..7021)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7036..7057
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7345..7933)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8104..8964)
                     /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(8965..9069)
                     /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 ggatccacta gtccagtgtg gtggaattcg ccaccatgaa ccaccagcag
      721 cagcagcagc agcagaaagc gggcgagcag cagttgagcg agcccgagga catggagatg
      781 gaagcgggag atacagatga cccaccaaga attactcaga accctgtgat caatgggaat
      841 gtggccctga gtgatggaca caacaccgcg gaggaggaca tggaggatga caccagttgg
      901 cgctccgagg caacctttca gttcactgtg gagcgcttca gcagactgag tgagtcggtc
      961 cttagccctc cgtgttttgt gcgaaatctg ccatggaaga ttatggtgat gccacgcttt
     1021 tatccagaca gaccacacca aaaaagcgta ggattctttc tccagtgcaa tgctgaatct
     1081 gattccacgt catggtcttg ccatgcacaa gcagtgctga agataataaa ttacagagat
     1141 gatgaaaagt cgttcagtcg tcgtattagt catttgttct tccataaaga aaatgattgg
     1201 ggattttcca attttatggc ctggagtgaa gtgaccgatc ctgagaaagg atttatagat
     1261 gatgacaaag ttacctttga agtctttgta caggcggatg ctccccatgg agttgcgtgg
     1321 gattcaaaga agcacacagg ctacgtcggc ttaaagaatc agggagcgac ttgttacatg
     1381 aacagcctgc tacagacgtt atttttcacg aatcagctac gaaaggctgt gtacatgatg
     1441 ccaaccgagg gggatgattc gtctaaaagc gtccctttag cattacaaag agtgttctat
     1501 gaattacagc atagtgataa acctgtagga acaaaaaagt taacaaagtc atttgggtgg
     1561 gaaactttag atagcttcat gcaacatgat gttcaggagc tttgtcgagt gttgctcgat
     1621 aatgtggaaa ataagatgaa aggcacctgt gtagagggca ccatacccaa attattccgc
     1681 ggcaaaatgg tgtcctatat ccagtgtaaa gaagtagact atcggtctga tagaagagaa
     1741 gattattatg atatccagct aagtatcaaa ggaaagaaaa atatatttga atcatttgtg
     1801 gattatgtgg cagtagaaca gctcgatggg gacaataaat acgacgctgg ggaacatggc
     1861 ttacaggaag cagagaaagg tgtgaaattc ctaacattgc caccagtgtt acatctacaa
     1921 ctgatgagat ttatgtatga ccctcagacg gaccaaaata tcaagatcaa tgataggttt
     1981 gaattcccag agcagttacc acttgatgaa tttttgcaaa aaacagatcc taaggaccct
     2041 gcaaattata ttcttcatgc agtcctggtt catagtggag ataatcatgg tggacattat
     2101 gtggtttatc taaaccccaa aggggatggc aaatggtgta aatttgatga cgacgtggtg
     2161 tcaaggtgta ctaaagagga agcaattgag cacaattatg ggggtcacga tgacgacctg
     2221 tctgttcgac actgcactaa tgcttacatg ttagtctaca tcagggaatc aaaactgagt
     2281 gaagttttac aggcggtcac cgaccatgat attcctcagc agttggtgga gcgattacaa
     2341 gaagagaaaa ggatcgaggc tcagaagcgg aaggagcggc aggaagccca tctctatatg
     2401 caagtgcaga tagtcgcaga ggaccagttt tgtggccacc aagggaatga catgtacgat
     2461 gaagaaaaag tgaaatacac tgtgttcaaa gtattgaaga actcctcgct tgctgagttt
     2521 gttcagagcc tctctcagac catgggattt ccacaagatc aaattcgatt gtggcccatg
     2581 caagcaagga gtaatggaac aaaacgacca gcaatgttag ataatgaagc cgacggcaat
     2641 aaaacaatga ttgagctcag tgataatgaa aacccttgga caatattcct ggaaacagtt
     2701 gatcccgagc tggctgctag tggagcgacc ttacccaagt ttgataaaga tcatgatgta
     2761 atgttatttt tgaagatgta tgatcccaaa acgcggagct tgaattactg tgggcatatc
     2821 tacacaccaa tatcctgtaa aatacgtgac ttgctcccag ttatgtgtga cagagcagga
     2881 tttattcaag atactagcct tatcctctat gaggaagtta aaccgaattt aacagagaga
     2941 attcaggact atgacgtgtc tcttgataaa gcccttgatg aactaatgga tggtgacatc
     3001 atagtatttc agaaggatga ccctgaaaat gataacagtg aattacccac cgcaaaggag
     3061 tatttccgag atctctacca ccgcgttgat gtcattttct gtgataaaac aatccctaat
     3121 gatcctggat ttgtggttac gttatcaaat agaatgaatt attttcaggt tgcaaagaca
     3181 gttgcacaga ggctcaacac agatccaatg ttgctgcagt ttttcaagtc tcaaggttat
     3241 agggatggcc caggtaatcc tcttagacat aattatgaag gtactttaag agatcttcta
     3301 cagttcttca agcctagaca acctaagaaa ctttactatc agcagcttaa gatgaaaatc
     3361 acagactttg agaacaggcg aagttttaaa tgtatatggt taaacagcca atttagggaa
     3421 gaggaaataa cactatatcc agacaagcat gggtgtgtcc gggacctgtt agaagaatgt
     3481 aaaaaggccg tggagcttgg ggagaaagca tcagggaaac ttaggctgct agaaattgta
     3541 agctacaaaa tcattggtgt tcatcaagaa gatgaactat tagaatgttt atctcctgca
     3601 acgagccgga cgtttcgaat agaggaaatc cctttggacc aggtggacat agacaaagag
     3661 aatgagatgc ttgtcacagt ggcgcatttc cacaaagagg tcttcggaac gttcggaatc
     3721 ccgtttttgc tgaggataca ccagggcgag cattttcgag aagtgatgaa gcgaatccag
     3781 agcctgctgg acatccagga gaaggagttt gagaagttta aatttgcaat tgtaatgatg
     3841 ggccgacacc agtacataaa tgaagacgag tatgaagtaa atttgaaaga ctttgagcca
     3901 cagcccggta atatgtctca tcctcggcct tggctagggc tcgaccactt caacaaagcc
     3961 ccaaagagga gtcgctacac ttaccttgaa aaggccatta aaatccataa cctcgagcac
     4021 catcaccatc accatcacca ttagggcccg atatctcgac aatcaacctc tggattacaa
     4081 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     4141 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     4201 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     4261 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     4321 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     4381 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     4441 ggtgttgtcg gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat
     4501 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     4561 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     4621 tcggatctcc ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg
     4681 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     4741 cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     4801 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     4861 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     4921 tgccatagca gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     4981 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     5041 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     5101 ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct
     5161 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     5221 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     5281 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat
     5341 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     5401 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5461 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     5521 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     5581 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5641 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     5701 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     5761 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     5821 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     5881 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     5941 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     6001 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     6061 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     6121 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     6181 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     6241 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     6301 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     6361 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     6421 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     6481 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     6541 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     6601 ggactctggg gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     6661 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     6721 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     6781 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     6841 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     6901 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     6961 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     7021 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     7081 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     7141 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     7201 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     7261 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     7321 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     7381 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     7441 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     7501 gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct
     7561 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     7621 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     7681 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7741 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     7801 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     7861 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     7921 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     7981 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     8041 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     8101 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     8161 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     8221 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     8281 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     8341 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     8401 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     8461 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     8521 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     8581 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     8641 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     8701 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     8761 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     8821 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     8881 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     8941 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     9001 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     9061 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     9121 gatcccctat ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     9181 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     9241 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagg
//
