LOCUS       V029642                 9094 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029642
VERSION     V029642
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"
     regulatory      655..664
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             664..729
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     CDS             733..3804
                     /label="NLRC4(NM_001199138)"
                     /note="NLRC4(NM_001199138)"
                     /gene="NLRC4"
     misc_feature    3870..4458
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4524..4572
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4774..5202
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5216..5545
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5612..6406
                     /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    6582..6703
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6752..6768)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6776..6792
                     /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(6800..6830)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6845..6866
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7154..7742)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7913..8773)
                     /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(8774..8878)
                     /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 ttaagccacc
      661 atggactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac
      721 gatgacaagc ttatgaattt cataaaggac aatagccgag cccttattca aagaatggga
      781 atgactgtta taaagcaaat cacagatgac ctatttgtat ggaatgttct gaatcgcgaa
      841 gaagtaaaca tcatttgctg cgagaaggtg gagcaggatg ctgctagagg gatcattcac
      901 atgattttga aaaagggttc agagtcctgt aacctctttc ttaaatccct taaggagtgg
      961 aactatcctc tatttcagga cttgaatgga caaagtcttt ttcatcagac atcagaagga
     1021 gacttggacg atttggctca ggatttaaag gacttgtacc ataccccatc ttttctgaac
     1081 ttttatcccc ttggtgaaga tattgacatt atttttaact tgaaaagcac cttcacagaa
     1141 cctgtcctgt ggaggaagga ccaacaccat caccgcgtgg agcagctgac cctgaatggc
     1201 ctcctgcagg ctcttcagag cccctgcatc attgaagggg aatctggcaa aggcaagtcc
     1261 actctgctgc agcgaattgc catgctctgg ggctccggaa agtgcaaggc tctgaccaag
     1321 ttcaaattcg tcttcttcct ccgtctcagc agggcccagg gtggactttt tgaaaccctc
     1381 tgtgatcaac tcctggatat acctggcaca atcaggaagc agacattcat ggccatgctg
     1441 ctgaagctgc ggcagagggt tcttttcctt cttgatggct acaatgaatt caagccccag
     1501 aactgcccag aaatcgaagc cctgataaag gaaaaccacc gcttcaagaa catggtcatc
     1561 gtcaccacta ccactgagtg cctgaggcac atacggcagt ttggtgccct gactgctgag
     1621 gtgggggata tgacagaaga cagcgcccag gctctcatcc gagaagtgct gatcaaggag
     1681 cttgctgaag gcttgttgct ccaaattcag aaatccaggt gcttgaggaa tctcatgaag
     1741 acccctctct ttgtggtcat cacttgtgca atccagatgg gtgaaagtga gttccactct
     1801 cacacacaaa caacgctgtt ccataccttc tatgatctgt tgatacagaa aaacaaacac
     1861 aaacataaag gtgtggctgc aagtgacttc attcggagcc tggaccactg tggagaccta
     1921 gctctggagg gtgtgttctc ccacaagttt gatttcgaac tgcaggatgt gtccagcgtg
     1981 aatgaggatg tcctgctgac aactgggctc ctctgtaaat atacagctca aaggttcaag
     2041 ccaaagtata aattctttca caagtcattc caggagtaca cagcaggacg aagactcagc
     2101 agtttattga cgtctcatga gccagaggag gtgaccaagg ggaatggtta cttgcagaaa
     2161 atggtttcca tttcggacat tacatccact tatagcagcc tgctccggta cacctgtggg
     2221 tcatctgtgg aagccaccag ggctgttatg aagcacctcg cagcagtgta tcaacacggc
     2281 tgccttctcg gactttccat cgccaagagg cctctctgga gacaggaatc tttgcaaagt
     2341 gtgaaaaaca ccactgagca agaaattctg aaagccataa acatcaattc ctttgtagag
     2401 tgtggcatcc atttatatca agagagtaca tccaaatcag ccctgagcca agaatttgaa
     2461 gctttctttc aaggtaaaag cttatatatc aactcaggga acatccccga ttacttattt
     2521 gacttctttg aacatttgcc caattgtgca agtgccctgg acttcattaa actggacttt
     2581 tatgggggag ctatggcttc atgggaaaag gctgcagaag acacaggtgg aatccacatg
     2641 gaagaggccc cagaaaccta cattcccagc agggctgtat ctttgttctt caactggaag
     2701 caggaattca ggactctgga ggtcacactc cgggatttca gcaagttgaa taagcaagat
     2761 atcagatatc tggggaaaat attcagctct gccacaagcc tcaggctgca aataaagaga
     2821 tgtgctggtg tggctggaag cctcagtttg gtcctcagca cctgtaagaa catttattct
     2881 ctcatggtgg aagccagtcc cctcaccata gaagatgaga ggcacatcac atctgtaaca
     2941 aacctgaaaa ccttgagtat tcatgaccta cagaatcaac ggctgccggg tggtctgact
     3001 gacagcttgg gtaacttgaa gaaccttaca aagctcataa tggataacat aaagatgaat
     3061 gaagaagatg ctataaaact agctgaaggc ctgaaaaacc tgaagaagat gtgtttattt
     3121 catttgaccc acttgtctga cattggagag ggaatggatt acatagtcaa gtctctgtca
     3181 agtgaaccct gtgaccttga agaaattcaa ttagtctcct gctgcttgtc tgcaaatgca
     3241 gtgaaaatcc tagctcagaa tcttcacaat ttggtcaaac tgagcattct tgatttatca
     3301 gaaaattacc tggaaaaaga tggaaatgaa gctcttcatg aactgatcga caggatgaac
     3361 gtgctagaac agctcaccgc actgatgctg ccctggggct gtgacgtgca aggcagcctg
     3421 agcagcctgt tgaaacattt ggaggaggtc ccacaactcg tcaagcttgg gttgaaaaac
     3481 tggagactca cagatacaga gattagaatt ttaggtgcat tttttggaaa gaaccctctg
     3541 aaaaacttcc agcagttgaa tttggcggga aatcgtgtga gcagtgatgg atggcttgcc
     3601 ttcatgggtg tatttgagaa tcttaagcaa ttagtgtttt ttgactttag tactaaagaa
     3661 tttctacctg atccagcatt agtcagaaaa cttagccaag tgttatccaa gttaactttt
     3721 ctgcaagaag ctaggcttgt tgggtggcaa tttgatgatg atgatctcag tgttattaca
     3781 ggtgctttta aactagtaac tgcttaagaa ttctgcagat atccagcaca gtggcggccg
     3841 ctcgagtcta gagggcccga tatctcgaca atcaacctct ggattacaaa atttgtgaaa
     3901 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     3961 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     4021 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     4081 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     4141 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     4201 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     4261 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     4321 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     4381 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     4441 tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga aggagacaat
     4501 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg
     4561 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     4621 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     4681 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag
     4741 atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     4801 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     4861 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     4921 taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     4981 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     5041 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     5101 tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt
     5161 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     5221 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     5281 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     5341 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     5401 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     5461 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     5521 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     5581 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     5641 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     5701 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     5761 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     5821 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     5881 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     5941 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     6001 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     6061 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     6121 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     6181 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     6241 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     6301 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     6361 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     6421 ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     6481 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     6541 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     6601 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     6661 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     6721 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     6781 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     6841 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6901 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6961 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     7021 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     7081 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     7141 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     7201 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     7261 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     7321 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     7381 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     7441 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7501 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7561 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     7621 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7681 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7741 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7801 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7861 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7921 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7981 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     8041 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     8101 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     8161 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     8221 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     8281 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     8341 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     8401 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     8461 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     8521 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     8581 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     8641 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     8701 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     8761 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8821 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8881 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     8941 gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     9001 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     9061 cttgaccgac aattgcatga agaatctgct tagg
//
