LOCUS       V034316                 8539 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V034316
VERSION     V034316
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             706..3195
                     /label="WDR75(NM_032168)"
                     /note="WDR75(NM_032168)"
                     /gene="WDR75"
     CDS             3202..3291
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    3315..3903
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3969..4017
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4219..4647
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4661..4990
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5057..5851
                     /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    6027..6148
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6197..6213)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6221..6237
                     /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(6245..6275)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6290..6311
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6599..7187)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7358..8218)
                     /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(8219..8323)
                     /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 ggatccacta gtccagtgtg gtggaattcg ccaccatggt ggaggaggag
      721 aacatccgcg tggttcgttg tggcggcagc gagttgaact ttaggagagc tgtgttctct
      781 gcagattcta agtatatctt ctgtgtctct ggagactttg ttaaagttta cagcacagtt
      841 acagaagagt gtgtacacat actgcatgga cacagaaatc tggtgactgg aatccagctt
      901 aaccccaaca accatctaca gctgtattct tgttcccttg atggcacaat taaactgtgg
      961 gactatatag atggcatctt aataaagact ttcatagttg gatgtaaact tcatgccctc
     1021 tttactcttg cccaagctga ggattctgtc tttgttatag tgaataaaga aaaaccagat
     1081 atatttcagc tggtttcagt gaaactgcca aaatcctcaa gccaggaagt agaagccaag
     1141 gagctgtcct ttgttttgga ttacataaac cagtcaccca agtgcattgc ctttggaaac
     1201 gagggagtat atgttgctgc agtacgggaa ttttacttgt ctgtttattt tttcaaaaag
     1261 aaaacaacat caaggtttac tttatcatca tcaagaaata agaagcatgc taaaaacaat
     1321 tttacatgtg tagcatgtca cccaacggaa gactgcatcg catctggtca catggatggc
     1381 aaaattcgtc tttggaggaa tttttatgat gataagaaat atacgtacac atgtttacat
     1441 tggcaccatg atatggttat ggatttggct ttttcagtga caggcaccag tctgctgagt
     1501 ggcggtcgtg aatctgtact tgtagagtgg cgcgatgcaa cagagaagaa taaggagttt
     1561 ctcccgcgtt taggagctac tattgaacat atctcagtct cgcctgcagg agatttattc
     1621 tgcacttctc actctgataa taagataata attattcacc gaaaccttga agcatccgca
     1681 gtaattcaag gcctagtgaa agataggagt atcttcactg gtttgatgat tgatccaaga
     1741 actaaagctt tggttttgaa tggaaaacct ggccacctgc agttttattc tctccagagt
     1801 gataaacagt tatacaattt agatattata cagcaagaat atattaatga ttatggtctg
     1861 atccaaattg aactaacaaa ggctgcattt ggctgctttg gtaactggct tgcaacagtg
     1921 gaacagcggc aagaaaagga aactgagctt gaattgcaaa tgaaactgtg gatgtataat
     1981 aagaaaacac aagggtttat tcttaacact aaaattaaca tgccacacga agactgcatt
     2041 acagctctct gtttctgtaa tgcagaaaaa tctgaacagc ccaccttggt tacagctagc
     2101 aaagatggtt acttcaaagt atggatatta acagatgact ctgacatata caaaaaagct
     2161 gttggctgga cctgtgactt tgttggtagt tatcacaagt atcaagcaac taactgttgt
     2221 ttctccgaag atggttcttt actagcagtt agttttgagg aaatagtcac aatatgggat
     2281 tctgtaacat gggaacttaa atgtacattt tgccaacgag ctgggaaaat aaggcacctt
     2341 tgctttggga gattgacgtg ttcaaagtat ctacttggtg ctactgaaaa tggcattctt
     2401 tgctgttgga atctgctgag ctgtgcattg gagtggaatg caaaattaaa tgttagagtt
     2461 atggaacccg atcctaattc agagaatatt gctgcaatct ctcagtcttc agtgggttca
     2521 gacttgtttg tatttaaacc tagtgagcca aggccattgt atattcaaaa gggtatctcc
     2581 agagagaaag tccagtgggg agtgtttgtt ccacgagatg tccctgaatc cttcacctca
     2641 gaagcttacc agtggctaaa tagatcccag ttttacttcc taacaaaatc acagagttta
     2701 ttgacattca gtacaaagtc tccagaagaa aaactcacac caacaagcaa acagctgcta
     2761 gcagaagaaa gtcttcccac aaccccattt tatttcatat tgggaaaaca caggcaacag
     2821 caggatgaaa aactaaacga aactttagag aatgagctgg tacaactacc cttaacagaa
     2881 aacatacccg caattagtga gcttcttcac actccagccc atgtcctgcc atctgctgct
     2941 ttcctgtgct ccatgtttgt aaattcattg ctgctgtcta aagagactaa gagtgctaag
     3001 gaaattcctg aagatgtaga tatggaagaa gaaaaagaaa gtgaagattc agatgaagaa
     3061 aatgatttta ccgaaaaagt ccaggataca agtaacacag gtttaggaga agacattata
     3121 catcagttgt caaaatctga agaaaaagaa ctgagaaaat ttaggaaaat agactacagc
     3181 tggatagctg cccttctcga ggaacaaaaa ctaatatcag aggaagattt ggaacagaaa
     3241 ctaatcagtg aagaagattt agagcaaaag ttaatttccg aggaggactt atagtgaggg
     3301 cccgatatct cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     3361 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     3421 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     3481 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     3541 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     3601 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     3661 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct
     3721 ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     3781 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     3841 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     3901 cgcctggaaa cgggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     3961 cgctatgacg gcaataaaaa gacagaataa aacgcacggg tgttgggtcg tttgttcata
     4021 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     4081 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     4141 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcagatctg cgcagctggg
     4201 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4261 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     4321 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc
     4381 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     4441 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4501 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     4561 tctattcttt tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc
     4621 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     4681 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4741 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4801 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     4861 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     4921 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4981 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     5041 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     5101 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     5161 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     5221 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     5281 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     5341 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     5401 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     5461 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     5521 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     5581 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     5641 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     5701 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     5761 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     5821 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc gaaatgaccg
     5881 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     5941 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     6001 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     6061 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     6121 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     6181 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     6241 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     6301 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     6361 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     6421 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     6481 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     6541 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     6601 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     6661 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6721 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6781 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6841 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6901 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6961 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     7021 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     7081 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     7141 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     7201 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     7261 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     7321 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     7381 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     7441 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     7501 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     7561 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     7621 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     7681 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7741 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     7801 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     7861 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     7921 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     7981 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     8041 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     8101 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     8161 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     8221 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     8281 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     8341 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac
     8401 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
     8461 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
     8521 catgaagaat ctgcttagg
//
