LOCUS       V019479                 7586 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019479
VERSION     V019479
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..2242
                     /label="USP3(NM_006537)"
                     /note="USP3(NM_006537)"
                     /gene="USP3"
     CDS             2249..2338
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    2362..2950
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3016..3064
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3266..3694
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3708..4037
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4104..4898
                     /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    5074..5195
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5244..5260)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5268..5284
                     /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(5292..5322)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5337..5358
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5646..6234)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6405..7265)
                     /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(7266..7370)
                     /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 ccatggagtg tccacacctg agctccagcg tctgcattgc
      721 tccggactca gccaagttcc ccaacggctc cccgtcgtcc tggtgctgca gcgtgtgccg
      781 gtccaacaaa agcccttggg tctgtttgac ttgttcaagt gtccactgtg gaaggtatgt
      841 gaatggccat gcaaaaaaac attatgaaga tgcacaagta cctttaacca accataagaa
      901 atcagaaaag caagataaag ttcagcacac agtatgtatg gattgcagta gctacagtac
      961 atactgttat cgctgtgatg attttgtggt taatgacacc aagctgggac tggtacagaa
     1021 agtcagagaa cacttacaga acttggaaaa ctcagctttc acagctgaca ggcataagaa
     1081 aagaaaactt ttggaaaact caacactaaa cagcaagtta ttaaaagtaa atggaagcac
     1141 cactgccatt tgtgccacag gccttcggaa tttggggaac acatgtttca tgaatgccat
     1201 ccttcagtca ctcagtaaca ttgagcagtt ttgctgttat ttcaaagaac tgcccgccgt
     1261 ggagttaagg aatgggaaaa cagcaggaag gcggacatac cacaccagga gccaagggga
     1321 taacaatgtg tctttggtag aagagtttag aaagacactc tgtgctttat ggcaaggcag
     1381 ccagactgca tttagcccag agtccttatt ttatgttgtt tggaagatta tgccaaactt
     1441 taggggctat caacagcagg acgcccatga attcatgcgc taccttttgg accacctaca
     1501 cttggaactt cagggcggtt tcaacggtgt ttcccgctca gcaattctgc aggagaattc
     1561 tactctgtct gcaagtaaca agtgttgcat aaatggagca tctactgttg tcacggctat
     1621 attcggaggc attctccaaa atgaggttaa ctgcctcata tgtgggacag aatctagaaa
     1681 gtttgatcca ttcctagacc tttcattaga tattccaagt cagttcagaa gtaagcgctc
     1741 taagaatcaa gaaaatggac cagtttgttc gttacgagat tgtcttcgca gttttaccga
     1801 cttagaagaa cttgatgaga cagagttata tatgtgccat aaatgcaaaa agaaacaaaa
     1861 gtccacaaaa aagttttgga ttcaaaaact acccaaggtg ctatgcttac atttgaaaag
     1921 atttcattgg acagcatatt taagaaataa agttgataca tacgtagaat ttccactgag
     1981 aggcctagac atgaaatgct acttactaga gcctgagaac agtggcccgg agagctgcct
     2041 gtatgacctc gccgctgtgg tggtgcacca tggttccggg gttggttctg gacattacac
     2101 agcatacgca actcacgaag gccgctggtt ccacttcaat gacagtactg taacactgac
     2161 tgacgaggag actgtggtga aggcgaaggc ctacatcctt ttctacgtgg aacaccaggc
     2221 caaagctgga tcggataaac ttctcgagga acaaaaacta atatcagagg aagatttgga
     2281 acagaaacta atcagtgaag aagatttaga gcaaaagtta atttccgagg aggacttata
     2341 gtgagggccc gatatctcga caatcaacct ctggattaca aaatttgtga aagattgact
     2401 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     2461 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     2521 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     2581 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     2641 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     2701 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg
     2761 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     2821 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     2881 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     2941 gcctccccgc ctggaaacgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     3001 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt
     3061 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     3121 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     3181 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc agatctgcgc
     3241 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3301 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3361 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3421 ggcatccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3481 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3541 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3601 atctcggtct attcttttga tttataaggg attttgggga tttcggccta ttggttaaaa
     3661 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     3721 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3781 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3841 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3901 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3961 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4021 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     4081 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4141 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4201 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4261 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     4321 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4381 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4441 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4501 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4561 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4621 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4681 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4741 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4801 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4861 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa
     4921 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     4981 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     5041 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     5101 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     5161 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     5221 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     5281 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     5341 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5401 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5461 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5521 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5581 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5641 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5701 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5761 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5821 ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5881 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5941 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     6001 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     6061 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     6121 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6181 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6241 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6301 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6361 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6421 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6481 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     6541 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     6601 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     6661 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6721 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6781 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6841 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6901 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6961 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     7021 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     7081 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     7141 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7201 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7261 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7321 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7381 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtcgactc
     7441 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
     7501 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
     7561 acaattgcat gaagaatctg cttagg
//
