LOCUS       V047198                 7608 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V047198
VERSION     V047198
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2288
                     /label="Nek3(NM_001162947)"
                     /note="Nek3(NM_001162947)"
                     /gene="Nek3"
     CDS             2295..2360
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2384..2972
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3038..3086
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3288..3716
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3730..4059
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4126..4920
                     /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    5096..5217
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5266..5282)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5290..5306
                     /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(5314..5344)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5359..5380
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5668..6256)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6427..7287)
                     /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(7288..7392)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgga caactacaca gtcctgagag
      781 tgattggcca gggctccttc ggcagggctc ttttggttct gcaggaaagc agcaatcaga
      841 catttgccat gaaggaaatc agactgctca agtctgatac acagacttct aggaaagagg
      901 ctgttctgct cgccaaaatg aagcacccca atattgttgc ctttaaggag tcatttgaag
      961 ctgaagggta cctgtacatt gtgatggagt attgtgacgg aggggatctg atgcagagga
     1021 ttaaacagca aaaagggaag ttattccctg aagacacgat acttaattgg tttatacaaa
     1081 tttgtcttgg agtaaatcac attcataaga gacgtgtttt acacagagat atcaagtcga
     1141 aaaatgtctt cctcacccac aatggaaaag tgaaattggg agattttggc tctgcccgtc
     1201 ttctctccag tcccatggcg tttgcatgca cgtacgtggg aacaccttat tatgtgcctc
     1261 cagaaatctg ggagaacctg ccttacaaca ataaaagtga catctggtcc ttgggatgca
     1321 ttctgtatga actctgcgct ctgaagcatc cattccaggc aaatagttgg aaaaacctga
     1381 tcctgaaaat atgccagggg cccatccacc cgctgccggc tctgtactcc tgcaagcttc
     1441 agggcctggt caagcagatg ctgaaaagga acccttccca ccgcccctcc gccacaaccc
     1501 tgctctgcag gggtagcttg gctccactgg tcccgaagtg cttgccccct cagatcatca
     1561 gagaatatgg agaacagata ttagatgaaa tcaaaatatc cacgcctaag aatatgaaaa
     1621 aacaggactc caacagagtc aggagagctc tgggagaagc aaactctgcc tctatgcaag
     1681 aggaagaacg tggtagaaag tgtagccata ctgaattgga aagcactggt acaaccccag
     1741 ctgggaatgc attggaaaga gcagccagag gaaatccaga atcaggtaac ccacaggagc
     1801 acgggaggca taccagtccg gcaagtcctc acaggccgtg gtgggagaga cacggtccca
     1861 gttccaatgt cgaagctttg gagaaagcat ctatccttac ctccagtttc acggcagagg
     1921 atgacagagg tgggtctgtg ataaagtatg aagaaaatgc ccgaaggcag tgggtcagag
     1981 agccccctga ggccttgctg tcaatgctga aggacgctga tctcagtcag gcatttcaga
     2041 catacacaat atatagacca ggtgcagaag gattcctgaa aggtcccctt tctgaagaca
     2101 cggcatcgga cagcgtcgac ggtgatctcg attctgtcat gttagaccca gagagatttg
     2161 aacctcggct ggatgaagaa gacacggact ttgaggaaga taacgaaaac cctgattggg
     2221 tgtcagaact aaagaagcac gtaggatacg gggatggacc tggtggacaa ctcctcggag
     2281 aacgtgccgg atccgactac aaagaccatg acggtgatta taaagatcat gacatcgact
     2341 acaaggatga cgatgacaag tagtgagggc ccgatatctc gacaatcaac ctctggatta
     2401 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     2461 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     2521 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     2581 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     2641 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     2701 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     2761 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     2821 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     2881 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     2941 gagtcggatc tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca
     3001 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     3061 acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     3121 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     3181 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     3241 ccctgccata gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag
     3301 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3361 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3421 tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca
     3481 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3541 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3601 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg
     3661 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3721 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3781 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3841 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3901 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3961 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     4021 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4081 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4141 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4201 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4261 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4321 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4381 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4441 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4501 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4561 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4621 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4681 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4741 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4801 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4861 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4921 gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     4981 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5041 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     5101 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5161 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5221 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     5281 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5341 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5401 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5461 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5521 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5581 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5641 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5701 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5761 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5821 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     5881 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5941 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6001 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6061 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     6121 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6181 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6241 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6301 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6361 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6421 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6481 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6541 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6601 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6661 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6721 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6781 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6841 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6901 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6961 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7021 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7081 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7141 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7201 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7261 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7321 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7381 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     7441 cccgatcccc tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag
     7501 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     7561 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
