LOCUS       V020046                 9368 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020046
VERSION     V020046
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..4024
                     /label="OTUD4(NM_001366057)"
                     /note="OTUD4(NM_001366057)"
                     /gene="OTUD4"
     CDS             4031..4120
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    4144..4732
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4798..4846
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5048..5476
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5490..5819
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5886..6680
                     /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    6856..6977
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7026..7042)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7050..7066
                     /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(7074..7104)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7119..7140
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7428..8016)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8187..9047)
                     /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(9048..9152)
                     /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 ccatggaggc tgccgtcggc gtccccgacg gcggggacca
      721 gggcggcgcg gggccccgcg aggacgcgac gcccatggac gcctatctgc ggaaactggg
      781 cttgtatcgg aaactggtcg ccaaggacgg gtcgtgcctg ttccgggccg tggcggagca
      841 ggtattgcac tctcagtctc gccatgttga agtcagaatg gcctgtattc actatcttcg
      901 agagaacaga gagaaatttg aagcgtttat agaaggatca tttgaagaat atttaaagcg
      961 tttggaaaat ccacaggaat gggtaggaca agtggaaata agtgcccttt ctcttatgta
     1021 caggaaagat tttataattt atcgggaacc aaatgtttct ccttcacaag taacagaaaa
     1081 taattttcct gaaaaggtgt tactgtgttt ttcaaatgga aatcattatg atattgtgta
     1141 tcccataaag tataaagaaa gctctgctat gtgtcagtct ctcctttatg aattgctgta
     1201 tgagaaggta tttaaaactg atgttagtaa aattgtgatg gaactagaca cgttggaagt
     1261 agctgatgaa gataacagtg aaatatcaga ttcagaggat gacagttgca agagtaagac
     1321 tgctgctgct gctgctgatg tgaatggatt taaacctttg tcaggcaatg agcagctgaa
     1381 gaacaatggg aactctacta gcctgccttt gtctagaaag gttcttaagt cactcaatcc
     1441 tgcagtctat agaaatgtgg aatatgaaat ttggctgaag tctaaacaag ctcagcaaaa
     1501 acgtgattat tccattgctg ctggcttaca atatgaagtt ggagacaaat gtcaagttag
     1561 gttggatcac aatggaaaat ttttgaatgc agatgttcaa ggaattcatt ctgagaatgg
     1621 accagttttg gttgaagaac tgggaaagaa gcacacatca aagaacctca aggcacctcc
     1681 cccagaaagc tggaacacag tgtcagggaa gaagatgaaa aaaccttcca cttctggaca
     1741 aaatttccat tctgatgtgg attacagagg gccaaagaat ccaagcaagc caataaaagc
     1801 cccatcagca ctacctcctc gactgcagca tccttcagga gtaagacaac atgcgttctc
     1861 tagtcattct tcagggtcac agtctcagaa attctccagt gagcacaaaa atcttagccg
     1921 gacaccttca cagatcataa gaaaacctga tcgtgaaaga gttgaggatt ttgatcacac
     1981 aagtcgagaa tctaactatt tcggcctttc cccagaagag cgcagagaga agcaagctat
     2041 agaagaatcc cgtttactct atgagattca gaacagagat gaacaggctt tcccagccct
     2101 ttccagctca tcagtcaatc agtcagcttc tcagagtagc aatccatgtg tccagagaaa
     2161 atcatcacat gtaggtgata gaaaaggaag caggcggaga atggatacag aagaacgaaa
     2221 agacaaagac tctattcatg gacatagtca gttggataaa agacccgaac caagcacatt
     2281 ggagaatatt actgatgata aatatgcaac agtttcatca ccatcaaagt caaagaagtt
     2341 agagtgccct tctcctgcgg aacaaaagcc agcagaacat gtgtctttgt caaatccagc
     2401 tccccttcta gtttctccag aggtacatct aactcctgcg gtgccttctt taccagccac
     2461 tgtgccagcc tggccaagtg aacctacaac ttttggacca acaggtgtcc ctgctccaat
     2521 tcccgttttg tcagtgacac agactttgac cactggacct gattcagctg tatcccaagc
     2581 tcatttaaca ccctctccag ttcctgtgtc aatacaggca gttaaccagc ccttgatgcc
     2641 tttgcctcag acattgagcc tttatcaaga cccactctat cctgggtttc cttgtaatga
     2701 aaagggagat cgagccattg taccacctta ttcactgtgt cagactgggg aggacctacc
     2761 taaagataag aatattcttc gattcttctt caatcttggt gtgaaggcat acagttgtcc
     2821 tatgtgggcc ccacattctt acctgtaccc tctgcaccag gcctacctgg cagcctgcag
     2881 gatgtaccca aaggtccctg tccctgttta tcctcataat ccctggttcc aagaggctcc
     2941 tgctgctcag aatgaaagtg attgtacctg tactgatgcc cactttccta tgcagactga
     3001 ggccagtgtt aatggtcaaa tgccacagcc agagattgga ccgccgacat tttcttcacc
     3061 tctggttatc cctccatctc aggtgtctga aagtcatgga caattgtctt accaggctga
     3121 tcttgaatct gagacccctg ggcagcttct gcatgctgat tatgaagagt cactaagtgg
     3181 caagaatatg ttcccccagc catcttttgg acccaatcca ttcttaggcc cagttcctat
     3241 tgcacctcct ttctttcctc atgtttggta tgggtaccct tttcagggat tcatagaaaa
     3301 tccagtaatg aggcagaata ttgtcctgcc ctctgatgaa aaaggagaat tggatctgtc
     3361 tctggaaaat ctggatctgt ctaaagattg tggttcagtt tcaacagtag atgagtttcc
     3421 agaagccagg ggtgaacatg tacattctct ccctgaagca agtgtgagca gtaagccgga
     3481 cgaaggccgg acagagcaat cttcccagac acgaaaggca gatacggcat tggcttccat
     3541 ccctcctgta gcagagggaa aggctcatcc tcccactcag attctaaaca gagagagaga
     3601 aactgtgcct gttgaacttg aacctaaaag gaccattcaa agcctgaaag aaaaaacaga
     3661 aaaagtaaaa gatcctaaga ctgctgctga tgtggtcagc cctggggcca actctgttga
     3721 tagcagagtg caaagaccaa aagaagagag ttcagaagat gaaaatgaag tgtctaatat
     3781 tttgagaagt ggtagatcca agcagttcta taatcaaact tatggaagca ggaagtacaa
     3841 aagtgattgg ggctattctg gtaggggtgg atatcaacat gtgagaagtg aggagtcctg
     3901 gaaaggacag ccaagcagaa gtcgggatga aggttatcag taccatcgaa atgtcagagg
     3961 gcgaccattt aggggagata ggaggagatc agggatggga gatggccata ggggacagca
     4021 cactctcgag gaacaaaaac taatatcaga ggaagatttg gaacagaaac taatcagtga
     4081 agaagattta gagcaaaagt taatttccga ggaggactta tagtgagggc ccgatatctc
     4141 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     4201 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     4261 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     4321 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     4381 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     4441 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     4501 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg
     4561 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     4621 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     4681 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaaac
     4741 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     4801 caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt
     4861 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     4921 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     4981 gccaacgtcg gggcggcagg ccctgccata gcagatctgc gcagctgggg ctctaggggg
     5041 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     5101 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     5161 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc
     5221 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     5281 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     5341 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     5401 gatttataag ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa
     5461 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     5521 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     5581 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     5641 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     5701 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     5761 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     5821 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
     5881 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     5941 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     6001 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     6061 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     6121 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     6181 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     6241 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     6301 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     6361 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     6421 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     6481 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     6541 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     6601 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     6661 ttcttgacga gttcttctga gcgggactct ggggttcgcg aaatgaccga ccaagcgacg
     6721 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     6781 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     6841 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     6901 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     6961 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     7021 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     7081 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     7141 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     7201 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     7261 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     7321 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     7381 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     7441 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     7501 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     7561 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     7621 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     7681 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     7741 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     7801 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     7861 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7921 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7981 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     8041 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     8101 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     8161 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     8221 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     8281 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     8341 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     8401 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     8461 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     8521 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     8581 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     8641 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     8701 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     8761 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     8821 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     8881 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     8941 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     9001 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     9061 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     9121 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     9181 gtcgacggat cgggagatct cccgatcccc tatggtcgac tctcagtaca atctgctctg
     9241 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     9301 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
     9361 tgcttagg
//
