LOCUS       V020429                 8978 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020429
VERSION     V020429
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             665..3637
                     /label="NEK9(NM_001329237)"
                     /note="NEK9(NM_001329237)"
                     /gene="NEK9"
     CDS             3644..3670
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3674..3700
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             3704..3730
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    3754..4342
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4408..4456
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4658..5086
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5100..5429
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5496..6290
                     /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    6466..6587
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6636..6652)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6660..6676
                     /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(6684..6714)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6729..6750
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7038..7626)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7797..8657)
                     /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(8658..8762)
                     /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 ttaagcttgc
      661 caccatgtcg gtgctgggcg agtacgagcg acactgcgat tccatcaact cggactttgg
      721 gagcgagtcc gggggttgcg gggactcgag tccggggcct agcgccagtc aggggccgcg
      781 agccggcggc ggcgcggcgg agcaggagga actgcactac atccccatcc gcgtcctggg
      841 ccgcggcgcc ttcggggaag ccacgctgta ccgccgcacc gaggatgact cactggttgt
      901 gtggaaggaa gtcgatttga cccggctgtc tgagaaggaa cgtcgtgatg ccttgaatga
      961 gattgttatt ctggcactgc tgcagcacga caacattatt gcctactaca atcacttcat
     1021 ggacaatacc acgctgctga ttgagctgga atattgtaat ggagggaacc tgtatgacaa
     1081 aatccttcgt cagaaggaca agttgtttga ggaagagatg gtggtgtggt acctatttca
     1141 gattgtttca gcagtgagct gcatccataa agctggaatc cttcatagag atataaagac
     1201 attaaatatt tttctgacca aggcaaacct gataaaactt ggagattatg gcctagcaaa
     1261 gaaacttaat tctgagtatt ccatggctga gacgcttgtg ggaaccccat attacatgtc
     1321 tccagagctc tgtcaaggag taaagtacaa tttcaagtct gatatctggg cagttggctg
     1381 cgtcattttt gaactgctta ccttaaagag gacgtttgat gctacaaacc cacttaacct
     1441 gtgtgtgaag atcgtgcaag gaattcgggc catggaagtt gactctagcc agtactcttt
     1501 ggaattgatc caaatggttc attcgtgcct tgaccaggat cctgagcaga gacctactgc
     1561 agatgaactt ctagatcgcc ctcttctcag gaaacgcagg agagagatgg aggaaaaagt
     1621 cactctgctt aatgcaccta caaagagacc aaggtcaagc actgtgactg aagcacccat
     1681 tgctgtagta acatcacgaa ccagtgaagt ctatgtttgg ggtggtggaa aatccacccc
     1741 ccagaaactg gatgttatca agagtggctg tagtgcccgg caggtctgtg cagggaatac
     1801 ccactttgct gtggtcacag tggagaagga actgtacact tgggtgaaca tgcaaggagg
     1861 cactaaactc catggtcagc tgggccatgg agacaaagcc tcctatcgac agccaaagca
     1921 tgtggaaaag ttgcaaggca aagctatccg tcaggtgtca tgtggtgatg atttcactgt
     1981 ctgtgtgact gatgagggtc agctctatgc cttcggatca gattattatg gctgcatggg
     2041 ggtggacaaa gttgctggcc ctgaagtgct agaacccatg cagctgaact tcttcctcag
     2101 caatccagtg gagcaggtct cctgtggaga taatcatgtg gtggttctga cacgaaacaa
     2161 ggaagtctat tcttggggct gtggcgaata tggtaggggt agccttgtct ctgtccactt
     2221 cttcacagga cgactgggtt tggattcaga agaggattat tatacaccac aaaaggtgga
     2281 tgttcccaag gccttgatta ttgttgcagt tcaatgtggc tgtgatggga catttctgtt
     2341 gacccagtca ggcaaagtgc tggcctgtgg actcaatgaa ttcaataagc tgggtctgaa
     2401 tcagtgcatg tcgggaatta tcaaccatga agcataccat gaagttccct acacaacgtc
     2461 ctttaccttg gccaaacagt tgtcctttta taagatccgt accattgccc caggcaagac
     2521 tcacacagct gctattgatg agcgaggccg gctgctgacc tttggctgca acaagtgtgg
     2581 gcagctgggc gttgggaact acaagaagcg tctgggaatc aacctgttgg ggggacccct
     2641 tggtgggaag caagtgatca gggtctcctg cggtgatgag tttaccattg ctgccactga
     2701 tgataatcac atttttgcct ggggcaatgg tggtaatggc cgcctggcaa tgacccccac
     2761 agagagacca catggctctg atatctgtac ctcatggcct cggcctattt ttggatctct
     2821 gcatcatgtc ccggacctgt cttgccgtgg atggcatacc attctcatcg ttgagaaagt
     2881 attgaattct aagaccatcc gttccaatag cagtggctta tccattggaa ctgtgtttca
     2941 gagctctagc ccgggaggag gcggcggggg cggcggtggt gaagaagagg acagtcagca
     3001 ggaatctgaa actcctgacc caagtggagg cttccgagga acaatggaag cagaccgagg
     3061 aatggaaggt ttaatcagtc ccacagaggc catggggaac agtaatgggg ccagcagctc
     3121 ctgtcctggc tggcttcgaa aggagctgga aaatgcagaa tttatcccca tgcctgacag
     3181 cccatctcct ctcagtgcag cgttttcaga atctgagaaa gataccctgc cctatgaaga
     3241 gctgcaagga ctcaaagtgg cctctgaagc tcctttggaa cacaaacccc aagtagaagc
     3301 ctcgtcacct cggctgaatc ctgcagtaac ctgtgctggg aagggaacac cactgactcc
     3361 tcctgcgtgt gcgtgcagct ctctgcaggt ggaggttgag agattgcagg gtctggtgtt
     3421 aaagtgtctg gctgaacaac agaagctaca gcaagaaaac ctccagattt ttacccaact
     3481 gcagaagttg aacaagaaat tagaaggagg gcagcaggtg gggatgcatt ccaaaggaac
     3541 tcagacagca aaggaagaga tggaaatgga tccaaagcct gacttagatt cagattcctg
     3601 gtgcctcctg ggaacagact cctgtagacc cagcctcctc gagtacccat acgacgtacc
     3661 tgattatgca acttatcctt atgatgtacc tgattatgct agctatcctt atgatgtccc
     3721 agactacgca tagtgagggc ccgatatctc gacaatcaac ctctggatta caaaatttgt
     3781 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     3841 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     3901 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     3961 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     4021 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     4081 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     4141 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
     4201 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     4261 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     4321 tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca cggaaggaga
     4381 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacgggt
     4441 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     4501 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     4561 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     4621 gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     4681 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4741 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4801 gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4861 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4921 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4981 acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg gatttcggcc
     5041 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     5101 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     5161 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     5221 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     5281 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     5341 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     5401 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     5461 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     5521 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     5581 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     5641 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
     5701 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     5761 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     5821 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     5881 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     5941 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     6001 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     6061 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     6121 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     6181 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     6241 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     6301 ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     6361 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     6421 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     6481 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     6541 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     6601 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     6661 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     6721 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     6781 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6841 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6901 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6961 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     7021 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     7081 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     7141 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     7201 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg
     7261 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     7321 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     7381 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     7441 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     7501 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     7561 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     7621 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7681 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7741 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7801 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7861 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7921 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7981 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     8041 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     8101 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     8161 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     8221 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     8281 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     8341 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     8401 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     8461 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     8521 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     8581 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     8641 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     8701 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     8761 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     8821 tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     8881 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     8941 aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
