LOCUS       V020300                 8159 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020300
VERSION     V020300
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..2815
                     /label="USP51(NM_201286)"
                     /note="USP51(NM_201286)"
                     /gene="USP51"
     CDS             2822..2911
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    2935..3523
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3589..3637
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3839..4267
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4281..4610
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4677..5471
                     /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    5647..5768
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5817..5833)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5841..5857
                     /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(5865..5895)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5910..5931
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6219..6807)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6978..7838)
                     /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(7839..7943)
                     /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 ccatggccca ggttcgagaa acttctttgc cctccggctc
      721 tggggtccgc tggatctccg gaggtggggg aggagcctct cctgaggagg cggttgagaa
      781 ggcggggaaa atggaggagg cggcggcggg ggctacgaag gcgtcttcga gacgtgaagc
      841 cgaggagatg aagctggagc cattacaaga gcgtgagccc gcgccggagg agaacttgac
      901 gtggagcagc agcggcggcg acgagaaggt gctcccttca atcccccttc gctgtcacag
      961 cagctcctcg cccgtttgcc cgcgccgcaa gccccgccct cggccccagc cccgggcccg
     1021 ctcccgcagc cagcctgggc tctcggcccc acccccgcct ccagcccggc ccccgccccc
     1081 gccgccaccc ccgcccccac ccgcaccgcg gcccagggcc tggcgtggat cccggcgcag
     1141 atcccggcct gggtccaggc ctcagacacg gagaagctgc tctggtgacc tagacgggtc
     1201 gggggatcct ggcggcttag gggactggtt gctggaggtc gagtttggtc agggtcccac
     1261 aggctgctct catgtggaga gctttaaagt aggtaagaac tggcagaaga acctgaggtt
     1321 gatctaccag cgtttcgttt ggagtgggac cccagagact aggaaacgta aagcaaagtc
     1381 atgcatctgt cacgtatgta gtacccatat gaacagactc cactcttgtc tctcctgtgt
     1441 cttttttggc tgcttcactg agaaacatat tcacaaacat gcagaaacaa agcagcacca
     1501 tttagctgta gacctttatc atggggtcat atattgcttc atgtgtaagg attatgtata
     1561 tgacaaagac atagaacaga ttgccaaaga aacaaaagaa aaaattttga gattattaac
     1621 ttccacctca acagatgttt ctcatcaaca gtttatgaca tcagggtttg aagacaagca
     1681 atcaacctgt gagacaaagg aacaggagcc aaaattggtg aaacccaaga aaaagagaag
     1741 aaaaaagtca gtctatactg taggcctgag agggctaatc aatcttggga acacttgttt
     1801 tatgaattgt attgtccagg cacttaccca tattcctcta ctgaaagatt tcttcctctc
     1861 tgacaagcac aaatgtataa tgacaagccc cagcttgtgt ctggtctgtg aaatgtcttc
     1921 gctttttcat gctatgtact ctgggagccg aactcctcac attccctata agttactgca
     1981 tctgatatgg atccatgcag aacatttagc agggtacagg cagcaggatg cccatgagtt
     2041 ccttattgca atattagacg tgctacatag acacagcaaa gatgatagtg gtgggcagga
     2101 ggccaataac cccaactgct gtaactgcat catagaccaa atctttacag gtggcctgca
     2161 atcagatgtc acatgtcaag cctgccatag tgtttctacc accatagacc catgctggga
     2221 catcagtttg gacttgcctg gctcttgtgc cacattcgat tcccagaacc cagagagggc
     2281 tgacagcaca gtgagcaggg atgaccacat accaggaatc ccctcactta cagactgtct
     2341 acagtggttt acaaggccag agcacctagg aagcagtgcc aaaatcaaat gcaatagttg
     2401 ccaaagctac caggagtcta ctaaacagct cacaatgaaa aaattaccca ttgtggcttg
     2461 ttttcatctc aagcggtttg agcatgtagg caaacagagg cgaaagatta atacctttat
     2521 ctcctttccc ttggagctgg acatgactcc gtttttggcc tctactaaag agagcagaat
     2581 gaaagaaggc cagccaccaa cagattgtgt gcccaatgag aataagtatt ccttgtttgc
     2641 agtgattaat caccatggaa ctttggaaag tggccactat accagcttca tccggcaaca
     2701 aaaggaccag tggttcagct gtgatgatgc catcatcacc aaggctacca ttgaggactt
     2761 actctacagt gaagggtatt tactgttcta tcacaaacag ggtctagaga aagacctcga
     2821 ggaacaaaaa ctaatatcag aggaagattt ggaacagaaa ctaatcagtg aagaagattt
     2881 agagcaaaag ttaatttccg aggaggactt atagtgaggg cccgatatct cgacaatcaa
     2941 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     3001 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     3061 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     3121 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     3181 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     3241 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     3301 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt
     3361 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     3421 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     3481 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa cgggggaggc
     3541 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     3601 gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     3661 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     3721 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     3781 ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac
     3841 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     3901 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3961 ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt
     4021 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     4081 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4141 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     4201 gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4261 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     4321 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     4381 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     4441 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4501 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     4561 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     4621 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     4681 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     4741 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     4801 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     4861 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4921 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4981 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     5041 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     5101 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     5161 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     5221 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     5281 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     5341 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     5401 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     5461 agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg
     5521 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     5581 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5641 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5701 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5761 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     5821 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     5881 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     5941 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     6001 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     6061 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     6121 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6181 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6241 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6301 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6361 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     6421 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6481 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6541 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6601 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     6661 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6721 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6781 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6841 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6901 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6961 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     7021 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     7081 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     7141 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     7201 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     7261 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     7321 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     7381 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     7441 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     7501 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     7561 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     7621 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     7681 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     7741 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     7801 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     7861 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     7921 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     7981 tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct gatgccgcat
     8041 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
     8101 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttagg
//
