LOCUS       V019459                 8768 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019459
VERSION     V019459
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..3424
                     /label="USP20(NM_006676)"
                     /note="USP20(NM_006676)"
                     /gene="USP20"
     CDS             3431..3520
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    3544..4132
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4198..4246
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4448..4876
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4890..5219
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5286..6080
                     /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    6256..6377
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6426..6442)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6450..6466
                     /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(6474..6504)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6519..6540
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6828..7416)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7587..8447)
                     /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(8448..8552)
                     /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 ccatggggga ctccagggac ctttgccctc accttgactc
      721 cataggagag gtgaccaaag aggacttgct gctcaaatct aagggaacct gtcagtcgtg
      781 tggggtcacc ggaccaaacc tatgggcctg tctgcaggtt gcctgcccct atgttggctg
      841 cggagaatcc tttgctgacc acagcaccat tcatgcacag gcaaaaaagc acaacttgac
      901 cgtgaacctg accacgttcc gactgtggtg ttacgcctgt gagaaggagg tattcctgga
      961 gcagcggctg gcagcccctc tgctgggctc ctcttccaag ttctctgaac aggactcccc
     1021 gccaccctcc caccctctga aagctgttcc tattgctgtg gctgatgaag gagagtctga
     1081 gtcagaggac gatgacctga aacctcgagg cctcacgggc atgaagaacc tcgggaactc
     1141 ctgctacatg aacgctgccc tgcaggccct gtccaattgc ccgccgctga ctcagttctt
     1201 cttggagtgt ggcggcctgg tgcgcacaga taagaagcca gccctgtgca agagctacca
     1261 gaagctggtc tctgaggtct ggcataagaa acggccaagc tacgtggtcc ccaccagtct
     1321 gtctcatggg atcaagttgg tcaacccaat gttccgaggc tatgcccagc aggacaccca
     1381 agagttcctt cgctgcctga tggaccagct gcacgaggag ctcaaggagc cggtggtggc
     1441 cacggtggcg ctgacggagg ctcgggactc agattcgagt gacacggatg agaaacggga
     1501 gggtgaccgg agcccatcag aagatgagtt cttgtcctgt gactcgagca gtgaccgggg
     1561 tgagggtgac gggcaggggc gtggcggggg cagctcgcag gccgagacgg agctgctgat
     1621 cccagatgag gcgggccgag ccatctctga gaaggagcgg atgaaggacc gcaagttctc
     1681 ctggggccag cagcgtacaa actcggagca agtggacgag gacgctgatg tggacactgc
     1741 catggctgcc cttgacgacc agcccgcgga ggcccagccc ccgtcaccac ggtcctccag
     1801 cccctgccgg acgccagagc cggacaatga tgctcaccta cgcagctcct ctcgcccctg
     1861 cagccccgtc caccaccacg agggccatgc caagctgtct agcagccccc ctcgtgcaag
     1921 ccccgtgagg atggcaccgt cgtacgtgct caagaaagcc caggtattga gtgctggcag
     1981 ccggaggcgg aaggagcagc gctaccgcag cgtcatctca gacatctttg acggctccat
     2041 tctcagcctt gtgcagtgtc tcacctgtga ccgggtatcc accacagtgg aaacgttcca
     2101 ggacttatca ctgcccattc ctggaaagga ggacctggcc aagctccatt cagccatcta
     2161 ccagaatgtg ccggccaagc caggcgcctg tggggacagc tatgccgccc agggctggct
     2221 ggccttcatt gtggagtaca tccgacggtt tgtggtatcc tgtaccccca gctggttttg
     2281 ggggcctgtc gtcaccctgg aagactgcct tgctgccttc tttgccgctg atgagttaaa
     2341 gggtgacaac atgtacagct gtgagcggtg taagaagctg cggaacggag tgaagtactg
     2401 caaagtcctg cggttgcccg agatcctgtg cattcaccta aagcgctttc ggcacgaggt
     2461 gatgtactca ttcaagatca acagccacgt ctccttcccc ctcgaggggc tcgacctgcg
     2521 ccccttcctt gccaaggagt gcacatccca gatcaccacc tacgacctcc tctcggtcat
     2581 ctgccaccac ggcacggcag gcagtgggca ctacatcgcc tactgccaga acgtgatcaa
     2641 tgggcagtgg tacgagtttg atgaccagta cgtcacagaa gtccacgaga cggtggtgca
     2701 gaacgccgag ggctacgtac tcttctacag gaagagcagc gaggaggcca tgcgggagcg
     2761 acagcaggtg gtgtccctgg ccgccatgcg ggagcccagc ctgctgcggt tctacgtgtc
     2821 ccgcgagtgg ctcaacaagt tcaacacctt cgcggagcca ggccccatca ccaaccagac
     2881 cttcctctgc tcccacggag gcatcccgcc ccacaaatac cactacatcg acgacctggt
     2941 ggtcatcctg ccccagaacg tctgggagca cctgtacaac agattcgggg gtggccccgc
     3001 cgtgaaccac ctgtacgtgt gctccatctg ccaggtggag atcgaggcac tggccaagcg
     3061 caggaggatc gagatcgaca ccttcatcaa gttgaacaag gccttccagg ccgaggagtc
     3121 gccgggcgtc atctactgca tcagcatgca gtggttccgg gagtgggagg cgttcgtcaa
     3181 ggggaaggac aacgagcccc ccgggcccat tgacaacagc aggattgcac aggtcaaagg
     3241 aagcggccat gtccagctga agcagggagc tgactacggg cagatttcgg aggagacctg
     3301 gacctacctg aacagcctgt atggaggtgg ccccgagatt gccatccgcc agagtgtggc
     3361 gcagccgctg ggcccagaga acctgcacgg ggagcagaag atcgaagccg agacgcgggc
     3421 cgtgctcgag gaacaaaaac taatatcaga ggaagatttg gaacagaaac taatcagtga
     3481 agaagattta gagcaaaagt taatttccga ggaggactta tagtgagggc ccgatatctc
     3541 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     3601 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     3661 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     3721 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     3781 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     3841 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     3901 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg
     3961 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     4021 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     4081 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaaac
     4141 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     4201 caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt
     4261 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     4321 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     4381 gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg ctctaggggg
     4441 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4501 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     4561 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc
     4621 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     4681 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     4741 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     4801 gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa
     4861 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     4921 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     4981 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     5041 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     5101 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     5161 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     5221 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     5281 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     5341 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     5401 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     5461 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     5521 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     5581 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     5641 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     5701 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     5761 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     5821 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     5881 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     5941 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     6001 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     6061 ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg
     6121 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     6181 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     6241 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     6301 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     6361 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     6421 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     6481 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     6541 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     6601 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6661 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6721 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6781 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6841 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6901 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6961 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     7021 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     7081 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     7141 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     7201 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     7261 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7321 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7381 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7441 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7501 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7561 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7621 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7681 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7741 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7801 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7861 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7921 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7981 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     8041 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     8101 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     8161 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     8221 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     8281 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     8341 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     8401 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     8461 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     8521 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     8581 gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca atctgctctg
     8641 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     8701 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
     8761 tgcttagg
//
