LOCUS       V029197                 8416 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029197
VERSION     V029197
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             964..3906
                     /label="USP15(NM_001252078)"
                     /note="USP15(NM_001252078)"
                     /gene="USP15"
     CDS             3913..3978
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    4016..4240
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4286..4714
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4728..5057
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5124..5918
                     /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    6092..6213
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6262..6278)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6286..6302
                     /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(6310..6340)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6355..6376
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6664..7249)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7420..8280)
                     /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(8281..8385)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatggcgg aaggcggagc ggcggatctg gacacccagc ggtctgacat cgcgacgctg
     1021 ctcaaaacct cgctccggaa aggggacacc tggtacctag tcgatagtcg ctggttcaaa
     1081 cagtggaaaa aatatgttgg ctttgacagt tgggacaaat accagatggg agatcaaaat
     1141 gtgtatcctg gacccattga taactctgga cttctcaaag atggtgatgc ccagtcactt
     1201 aaggaacacc ttattgatga attggattac atactgttgc caactgaagg ttggaataaa
     1261 cttgtcagct ggtacacatt gatggaaggt caagagccaa tagcacgaaa ggtggttgaa
     1321 cagggtatgt ttgtaaagca ctgcaaagta gaagtatatc tcacagaatt gaagctatgt
     1381 gaaaatggaa acatgaataa tgttgtaact cgaagattta gcaaagctga cacaatagat
     1441 acaattgaaa aggaaataag aaaaatcttc agtattccag atgaaaagga gaccagattg
     1501 tggaacaaat acatgagtaa cacatttgaa ccactgaata aaccagacag caccattcag
     1561 gatgctggtt tataccaagg acaggtatta gtgatagaac agaaaaatga agatggaaca
     1621 tggccaaggg gtccttctac tcctaagtcc ccaggtgcat ccaatttttc aactttacca
     1681 aagatctctc cttcatctct atcaaataat tataacaaca tgaacaacag aaatgtgaaa
     1741 aactcaaatt actgtcttcc atcatatacc gcttataaga actatgatta ttcggaacct
     1801 ggaagaaaca atgaacagcc aggcctctgt ggcctaagta acttgggaaa tacgtgtttc
     1861 atgaactcag ctattcagtg tttgagcaac acacctccac ttactgagta tttcctcaat
     1921 gataagtatc aagaagaact gaattttgac aatcccttag gaatgagagg tgaaatagct
     1981 aaatcttatg ccgaactgat caagcaaatg tggtctggaa agtttagcta cgtcacccca
     2041 agagccttta agacacaggt aggacgtttt gcacctcagt tctctggata tcagcagcaa
     2101 gactgtcaag aactgttagc tttcctatta gatggattac atgaggattt gaatagaatt
     2161 aggaaaaaac catatataca attaaaagat gcagatggaa ggccagataa ggtggttgcc
     2221 gaagaagcct gggaaaacca tttaaaacga aatgattcta tcatagtaga tatatttcat
     2281 ggccttttca aatcaacttt agtttgtcct gagtgtgcta agatttcagt aacatttgat
     2341 cctttttgtt acttgacact tccattgccc atgaaaaaag aacgcacctt ggaagtttac
     2401 ttagttagaa tggatccact taccaaacct atgcagtaca aagtggttgt ccccaaaatt
     2461 ggaaacatat tagatctttg tacagcattg tctgctttgt caggaatacc tgcagataag
     2521 atgatagtta ctgatatata caatcataga tttcacagaa tattcgctat ggatgaaaac
     2581 cttagtagta ttatggaacg ggatgatatt tatgtgtttg aaattaacat caataggaca
     2641 gaagatacag agcacgtgat tattcctgtt tgcctaagag aaaaattcag acactcgagt
     2701 tatacccacc atactggttc ttcacttttt ggtcagccct ttcttatggc tgtaccacga
     2761 aacaatactg aagacaaact ttataatctc ctgctcttga gaatgtgccg atatgtcaaa
     2821 atatctactg aaactgaaga aactgaagga tccctacact gctgtaagga ccaaaatatt
     2881 aatgggaatg gcccaaatgg catacatgaa gaaggctcac caagtgaaat ggaaacagat
     2941 gagccagatg atgaatccag ccaggatcaa gaacttccct cagagaatga aaacagtcag
     3001 tctgaagatt cagttggagg agataatgat tctgaaaatg gattatgtac tgaggatact
     3061 tgcaaaggtc aactcacggg acacaaaaaa cgattgttta cattccagtt caacaactta
     3121 ggcaatactg atatcaacta catcaaagat gataccaggc atataagatt tgatgatagg
     3181 cagcttaggc tagatgaaag atcttttctt gctttggatt gggatcctga tttgaaaaaa
     3241 agatattttg atgaaaatgc tgctgaggac tttgaaaaac atgaaagtgt ggagtataaa
     3301 cctcctaaaa aaccctttgt gaaattaaaa gattgcattg aactttttac aacaaaagaa
     3361 aagctaggtg ctgaagatcc ctggtattgt ccgaattgta aagaacatca gcaagccaca
     3421 aagaaattgg atttatggtc cctgcctcca gtacttgtag tacatctcaa gcgattttct
     3481 tacagtcgat acatgagaga caagttggat accttagttg attttcctat caatgacttg
     3541 gatatgtcgg aattcttaat taatccaaat gcaggtcctt gccgctataa tctgattgct
     3601 gtttccaacc actatggagg gatgggagga ggacactata ctgcttttgc aaaaaataaa
     3661 gatgatggaa aatggtacta ttttgatgac agtagtgtct ccactgcatc tgaagaccaa
     3721 attgtgtcca aagcagcata tgtactcttc taccagagac aagacacttt cagtggaact
     3781 ggcttttttc ctcttgaccg agaaactaaa ggtgcttcag ctgccactgg catcccatta
     3841 gaaagtgatg aagatagcaa tgataatgac aatgatatag aaaatgaaaa ctgtatgcac
     3901 actaacctcg aggactacaa agaccatgac ggtgattata aagatcatga catcgactac
     3961 aaggatgacg atgacaagta gtgagggccc gtttaaaccc gctgatcagc ctcgactgtg
     4021 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     4081 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     4141 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     4201 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     4261 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     4321 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     4381 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     4441 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4501 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4561 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     4621 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     4681 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     4741 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4801 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4861 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4921 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4981 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     5041 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     5101 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     5161 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     5221 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     5281 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     5341 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     5401 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     5461 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     5521 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     5581 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     5641 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     5701 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5761 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     5821 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5881 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     5941 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     6001 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     6061 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     6121 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     6181 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     6241 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     6301 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     6361 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6421 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6481 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6541 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6601 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6661 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6721 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6781 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6841 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6901 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6961 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     7021 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     7081 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     7141 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7201 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     7261 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     7321 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7381 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7441 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     7501 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     7561 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     7621 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     7681 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     7741 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     7801 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     7861 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     7921 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     7981 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     8041 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     8101 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     8161 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     8221 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     8281 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     8341 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8401 agtgccacct gacgtc
//
