LOCUS       V023164                 8094 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023164
VERSION     V023164
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..2774
                     /label="Nlrp10(NM_175532)"
                     /note="Nlrp10(NM_175532)"
                     /gene="Nlrp10"
     CDS             2781..2846
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2870..3458
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3524..3572
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3774..4202
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4216..4545
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4612..5406
                     /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    5582..5703
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5752..5768)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5776..5792
                     /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(5800..5830)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5845..5866
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6154..6742)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6913..7773)
                     /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(7774..7878)
                     /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 ctcggatccg ccaccatggc cttggcacgg gccaacagcc
      781 cccaggaggc actgctttgg gccttgaatg accttgagga gaacagtttc aagacgctga
      841 agttccactt acgagatgtg acccaatttc atctggcccg aggggaactg gagagtctga
      901 gtcaggtgga cctggcatca aaactgattt caatgtacgg agcacaggaa gccgtgagag
      961 tggtgagcag gagcttgctg gccatgaact tgatggagct tgtagactac ctcaaccagg
     1021 tttgtctgaa tgattacaga gaaatctacc gggagcatgt gcgctgcttg gaggagaggc
     1081 aagattgggg tgttaatagc agccacaata agctgctcct gatggccaca tccagttcag
     1141 ggggtcgcag atcaccttcc tgttctgacc tggaacagga gttggatcct gtcgatgtgg
     1201 agactttatt tgctccggag gcggagtcct actcaacccc gcccatagtg gtgatgcaag
     1261 gatctgcggg cactggaaag acaaccctgg tcaaaaaact ggtgcaggac tggtccaaag
     1321 ggaagctgta cccaggccag tttgattatg ttttctacgt gagctgcaga gaagtggtcc
     1381 tgttacccaa gtgtgacctg cccaacctca tctgctggtg ttgtggagat gatcaagccc
     1441 cagtcacaga gattctgcgg cagcctgggc ggctcctgtt catcctggat ggctatgatg
     1501 agttacagaa gtccagtcgt gccgagtgcg tacttcacat tctgatgagg agaagggagg
     1561 taccatgctc ccttctcatc accacgcggc ctccagcttt gcagagtctg gaacccatgt
     1621 tgggtgaacg gcgacatgtc cttgtcttag gcttctctga ggaggagagg gaaacatatt
     1681 tcagctcctg ttttacggat aaggagcaat tgaaaaatgc ccttgagttt gtgcaaaaca
     1741 atgctgttct ctataaagca tgccaggttc caggcatttg ctgggtggtc tgctcctggc
     1801 tgaagaagaa gatggcgaga ggtcaggaag tctcagagac acctagcaac agtacagaca
     1861 tcttcactgc ttatgtgtcc acctttctgc ccactgatgg caatggagac agctctgagc
     1921 ttaccaggca caaggtactg aagagtctgt gctccttggc tgcggagggg atgcggcacc
     1981 agaggctgct atttgaagaa gaggtcctca gaaagcatgg cttagacggc cccagcctta
     2041 ctgctttcct gaactgcatc gactaccgag cggggcttgg catcaagaaa ttctacagct
     2101 ttcgccacat cagcttccag gaatttttct atgccatgtc cttccttgtg aaggaggacc
     2161 agagtcagca gggagaggcc acacacaaag aagtggcaaa gctggtggat ccagagaacc
     2221 atgaagaggt gactctcagt ttgcagttct tattcgacat gttgaaaaca gagggcacct
     2281 tgagcttggg gctgaagttc tgcttcagaa tcgctccttc ggtaaggcag gatctgaagc
     2341 attttaaaga gcaaatagaa gccataaagt acaaaaggtc atgggatctg gaattctctc
     2401 tctatgattc taagataaag aagctcacac aaggtatcca gatgaaagat gtcattctca
     2461 atgtgcaaca tttggatgaa aagaaatctg acaagaagaa gtcagtttca gtgacaagca
     2521 gctttagtag cgggaaggta caaagtccat tcttgggaaa tgataaaagc accaggaaac
     2581 aaaagaaggc atctaatggg aaaagcagag gggcagagga accagccccg ggggttagaa
     2641 accgtaggct ggcaagcaga gaaaagggtc atatggagat gaatgataag gaggatggtg
     2701 gggtggagga acaggaggat gaggaaggac agacacttaa aaaggatgga gagatgatag
     2761 ataagatgaa tgggctcgag gactacaaag accatgacgg tgattataaa gatcatgaca
     2821 tcgactacaa ggatgacgat gacaagtagt gagggcccga tatctcgaca atcaacctct
     2881 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     2941 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     3001 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     3061 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     3121 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     3181 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3241 caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc
     3301 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3361 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3421 tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaaacgggg gaggctaact
     3481 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     3541 aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     3601 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     3661 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     3721 ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc
     3781 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3841 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3901 cggctttccc cgtcaagctc taaatcgggg catcccttta gggttccgat ttagtgcttt
     3961 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4021 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4081 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4141 tttggggatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4201 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4261 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4321 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4381 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4441 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     4501 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     4561 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     4621 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4681 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4741 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     4801 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4861 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4921 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4981 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     5041 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5101 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     5161 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5221 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5281 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5341 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5401 ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc
     5461 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5521 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5581 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5641 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5701 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     5761 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5821 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5881 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5941 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     6001 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6061 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6121 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6181 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6241 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6301 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     6361 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6421 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6481 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6541 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     6601 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6661 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6721 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6781 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6841 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6901 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6961 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7021 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7081 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7141 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7201 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7261 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7321 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7381 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7441 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7501 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7561 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7621 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7681 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7741 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7801 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7861 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     7921 agatctcccg atcccctatg gtcgactctc agtacaatct gctctgatgc cgcatagtta
     7981 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     8041 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagg
//
