LOCUS       V027509                 7826 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027509
VERSION     V027509
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             941..3316
                     /label="Nek4(NM_011849)"
                     /note="Nek4(NM_011849)"
                     /gene="Nek4"
     CDS             3323..3388
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3426..3650
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3696..4124
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4138..4467
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4534..5328
                     /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    5502..5623
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5672..5688)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5696..5712
                     /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(5720..5750)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5765..5786
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6074..6659)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6830..7690)
                     /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(7691..7795)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccgccacc atgccccagg ccgcatactg
      961 ctacatgcgg gtcgtgggca ggggtagtta cggggaggtg acgctcgtga agcatcggcg
     1021 agacggcaag cagtatgtca tcaaaaagct gaacctccga aatgcctcca gccgagagcg
     1081 tcgagctgct gagcaggaag ctcagctcct atctcagttg aagcacccca acattgtcac
     1141 ctacaaagaa tcatgggagg gaggagacgg tctgctctac attgtcatgg gcttctgcga
     1201 aggaggtgat ctgtaccgga agctcaagga gcagaaaggg cagcttctgc ctgagagtca
     1261 ggtggtggag tggtttgtcc agatcgccat ggccttgcag tatttacatg agaaacacat
     1321 ccttcatcga gatctgaaaa ctcagaatgt cttcttaaca agaacaaaca tcatcaaagt
     1381 gggggaccta ggaattgccc gcgtcttaga gaatcacggt gacatggcga gcaccctcat
     1441 tggcacaccc tattacatga gccctgagct gttctccaac aaaccctaca actataagtc
     1501 ggatgtttgg gctttgggat gctgtgtata tgaaatggcc accctgaagc acgcttttaa
     1561 tgcaaaagac atgaattctt tagtttatcg gattattgaa ggaaagttgc cgccaatgcc
     1621 aaaagtttac agcacagagc tggctgagct gataagaaca atgctgagca gaaggcctga
     1681 ggagaggccc tctgtgcgga gcatcctgag gcagccctac ataaaacacc acatctcctt
     1741 gttcttggag gccaccaagg caaaaacttc caaaaataat gttaaaaact gtgactccag
     1801 agccaagcct gttgctgctg tagtttccag aaaggaagaa tcaaatactg acgtgataca
     1861 ctaccagcca cgctcttctg agggctctgc tttgcatgtc atgggtgaag ataaatgttt
     1921 gtcccaagag aaaccagtgg acattggtcc cttgagatca ccggctagtc ttgaaggcca
     1981 cactggcaaa caagacatga acaatactgg ggagtcatgt gccacaatca gtaggataaa
     2041 tatcgatatc ttacctgcag aaaggaggga ctcagcgaac gctggtgtag ttcaggagag
     2101 ccaaccacaa cacgtggatg ccgctgatga agtagacagt cagtgcagta tttctcaaga
     2161 gaaggagagg ctacagggca ataccaagtc cagtgatcag cctggaaatc tccttcccag
     2221 acggtcctct gatggtggtg atggggaagg gagtgaactg gtgaagccat tgtatccctc
     2281 aaataaagac caaaagccag accaggatca agttactggt attatagaaa atcaggacag
     2341 catccaccca agatcccagc cacacagctc tatgtctgaa ccttctttgt cccggcagcg
     2401 aaggcagaaa aaaagggaac agactgccca tagtcgtaca aagagccagt tccaagaatt
     2461 accccctcgg cttttgcctt cttatcctgg tattgggaaa gtggatatca tagcaacaca
     2521 gcaaaatgat ggaaaccaag gtggacctgt ggctgggtgt gtaaacagtt caaggaccag
     2581 ctcaacagca tcagcaaagg atcgaccatt atcagccaga gagaggagac ggctaaagca
     2641 gtcacaggag gagatgctgc cctcaggccc tgcagttcag agaactccaa gtgcagttga
     2701 gccactgaaa ccacaggaag aagaccaacc catccctgcc caacgattct cttctgactg
     2761 cagcatcact cagatgaatc acacactccc gagggaaaag gagaagaggc tgatgcatgg
     2821 cctgtctgag gatgagttaa gctcttctac aagctcaact gataagtcag atggggattc
     2881 cagggagggg aagagtcata caaatgaaat gaaagacttg gtacagctga tgactcagac
     2941 tctgaggctg gaagctaagg agagctgtga agatctccag gtcctaaacc cagggtcaga
     3001 attcagactt catcgaaagt atcgagacac gctggtgctt catggcaagg ttgcagaaga
     3061 ggtggaaccc cactgtacgg agctgcctac gggtatcata ccaggttctg aaaagatcag
     3121 gagaatcgtt gaagtcttaa gagctgatgt aatccagggc ctggggattc aactcttaga
     3181 gcaggtgttt gatcttttgg gagaggaaga tgagttggag agagaggcac gattacagga
     3241 gcacatggga gacaagtata caacttactg tgtgaaagct cgccaattga aattttttga
     3301 agaaaatgtg agttttctcg aggactacaa agaccatgac ggtgattata aagatcatga
     3361 catcgactac aaggatgacg atgacaagta gtgagggccc gtttaaaccc gctgatcagc
     3421 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     3481 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     3541 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     3601 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     3661 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     3721 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3781 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3841 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3901 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3961 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4021 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     4081 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     4141 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4201 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4261 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4321 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4381 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     4441 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     4501 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     4561 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     4621 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     4681 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     4741 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4801 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4861 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4921 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4981 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     5041 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     5101 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     5161 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     5221 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     5281 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     5341 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     5401 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     5461 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     5521 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     5581 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     5641 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     5701 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     5761 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5821 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5881 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5941 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6001 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6061 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6121 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6181 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6241 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6301 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6361 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6421 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6481 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     6541 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6601 ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6661 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6721 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6781 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6841 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6901 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6961 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7021 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7081 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7141 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7201 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7261 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7321 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7381 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7441 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7501 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7561 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7621 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7681 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7741 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7801 ttccccgaaa agtgccacct gacgtc
//
