LOCUS       V022946                 9267 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022946
VERSION     V022946
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..3941
                     /label="NLRP12(NM_001277126)"
                     /note="NLRP12(NM_001277126)"
                     /gene="NLRP12"
     CDS             3954..4019
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    4043..4631
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4697..4745
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4947..5375
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5389..5718
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5785..6579
                     /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    6755..6876
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6925..6941)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6949..6965
                     /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(6973..7003)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7018..7039
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7327..7915)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8086..8946)
                     /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(8947..9051)
                     /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 ccaccatgct acgaaccgca ggcagggacg
      781 gcctctgtcg cctgtccacc tacttggaag aactcgaggc tgtggaactg aagaagttca
      841 agttatacct ggggaccgcg acagagctgg gagaaggcaa gatcccctgg ggaagcatgg
      901 agaaggccgg tcccctggaa atggcccagc tgctcatcac ccacttcggg ccagaggagg
      961 cctggaggtt ggctctcagc acctttgagc ggataaacag gaaggacctg tgggagagag
     1021 gacagagaga ggacctggtg agggataccc cacctggtgg cccgtcctca cttgggaacc
     1081 agtcaacatg ccttctggaa gtctctcttg tcactccaag aaaagatccc caggaaacct
     1141 acagggacta tgtccgcagg aaattccggc tcatggaaga ccgcaatgcg cgcctagggg
     1201 aatgtgtcaa cctcagccac cggtacaccc ggctcctgct ggtgaaggag cactcaaacc
     1261 ccatgcaggt ccagcagcag cttctggaca caggccgggg acacgcgagg accgtgggac
     1321 accaggctag ccccatcaag atagagaccc tctttgagcc agacgaggag cgccccgagc
     1381 caccgcgcac cgtggtcatg caaggcgcgg cagggatagg caagtccatg ctggcacaca
     1441 aggtgatgct ggactgggcg gacgggaagc tcttccaagg cagatttgat tatctcttct
     1501 acatcaactg cagggagatg aaccagagtg ccacggaatg cagcatgcaa gacctcatct
     1561 tcagctgctg gcctgagccc agcgcgcctc tccaggagct catccgagtt cccgagcgcc
     1621 tccttttcat catcgacggc ttcgatgagc tcaagccttc tttccacgat cctcagggac
     1681 cctggtgcct ctgctgggag gagaaacggc ccacggagct gcttcttaac agcttaattc
     1741 ggaagaagct gctccctgag ctatctttgc tcatcaccac acggcccacg gctttggaga
     1801 agctccaccg tctgctggag caccccaggc atgtggagat cctgggcttc tctgaggcag
     1861 aaaggaagga atacttctac aagtatttcc acaatgcaga gcaggcgggc caagtcttca
     1921 attacgtgag ggacaacgag cctctcttca ccatgtgctt cgtccccctg gtgtgctggg
     1981 tggtgtgtac ctgcctccag cagcagctgg agggtggggg gctgttgaga cagacgtcca
     2041 ggaccaccac tgcagtgtac atgctctacc tgctgagtct gatgcaaccc aagccggggg
     2101 ccccgcgcct ccagccccca cccaaccaga gagggttgtg ctccttggcg gcagatgggc
     2161 tctggaatca gaaaatccta tttgaggagc aggacctccg gaagcacggc ctagacgggg
     2221 aagacgtctc tgccttcctc aacatgaaca tcttccagaa ggacatcaac tgtgagaggt
     2281 actacagctt catccacttg agtttccagg aattctttgc agctatgtac tatatcctgg
     2341 acgaggggga gggcggggca ggcccagacc aggacgtgac caggctgttg accgagtacg
     2401 cgttttctga aaggagcttc ctggcactca ccagccgctt cctgtttgga ctcctgaacg
     2461 aggagaccag gagccacctg gagaagagtc tctgctggaa ggtctcgccg cacatcaaga
     2521 tggacctgtt gcagtggatc caaagcaaag ctcagagcga cggctccacc ctgcagcagg
     2581 gctccttgga gttcttcagc tgcttgtacg agatccagga ggaggagttt atccagcagg
     2641 ccctgagcca cttccaggtg atcgtggtca gcaacattgc ctccaagatg gagcacatgg
     2701 tctcctcgtt ctgtctgaag cgctgcagga gcgcccaggt gctgcacttg tatggcgcca
     2761 cctacagcgc ggacggggaa gaccgcgcga ggtgctccgc aggagcgcac acgctgttgg
     2821 tgcagctcag accagagagg accgttctgc tggacgccta cagtgaacat ctggcagcgg
     2881 ccctgtgcac caatccaaac ctgatagagc tgtctctgta ccgaaatgcc ctgggcagcc
     2941 ggggggtgaa gctgctctgt caaggactca gacaccccaa ctgcaaactt cagaacctga
     3001 ggctgaagag gtgccgcatc tccagctcag cctgcgagga cctctctgca gctctcatag
     3061 ccaataagaa tttgacaagg atggatctca gtggcaacgg cgttggattc ccaggcatga
     3121 tgctgctttg cgagggcctg cggcatcccc agtgcaggct gcagatgatt cagttgagga
     3181 agtgtcagct ggagtccggg gcttgtcagg agatggcttc tgtgctcggc accaacccac
     3241 atctggttga gttggacctg acaggaaatg cactggagga tttgggcctg aggttactat
     3301 gccagggact gaggcaccca gtctgcagac tacggacttt gtggctgaag atctgccgcc
     3361 tcactgctgc tgcctgtgac gagctggcct caactctcag tgtgaaccag agcctgagag
     3421 agctggacct gagcctgaat gagctggggg acctcggggt gctgctgctg tgtgagggcc
     3481 tcaggcatcc cacgtgcaag ctccagaccc tgcggttggg catctgccgg ctgggctctg
     3541 ccgcctgtga gggtctttct gtggtgctcc aggccaacca caacctccgg gagctggact
     3601 tgagtttcaa cgacctggga gactggggcc tgtggttgct ggctgagggg ctgcaacatc
     3661 ccgcctgcag actccagaaa ctgtggctgg atagctgtgg cctcacagcc aaggcttgtg
     3721 agaatcttta cttcaccctg gggatcaacc agaccttgac cgacctttac ctgaccaaca
     3781 acgccctagg ggacacaggt gtccgactgc tttgcaagcg gctgagccat cctggctgca
     3841 aactccgagt cctctggtta tttgggatgg acctgaataa aatgacccac agtaggttgg
     3901 cagcgcttcg agtaacaaaa ccttatttgg acattggctg cctcgagtct agagactaca
     3961 aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac gatgacaagt
     4021 agtgagggcc cgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     4081 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     4141 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     4201 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     4261 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     4321 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     4381 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     4441 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     4501 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     4561 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     4621 cgcctccccg cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa
     4681 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt
     4741 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     4801 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     4861 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg
     4921 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     4981 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     5041 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     5101 gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     5161 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     5221 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     5281 tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct attggttaaa
     5341 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     5401 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     5461 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     5521 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     5581 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     5641 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     5701 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     5761 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     5821 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     5881 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     5941 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     6001 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     6061 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     6121 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     6181 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     6241 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     6301 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     6361 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     6421 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     6481 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     6541 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga
     6601 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     6661 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     6721 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     6781 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     6841 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     6901 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     6961 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     7021 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     7081 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     7141 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     7201 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     7261 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     7321 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     7381 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     7441 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     7501 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     7561 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     7621 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     7681 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7741 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     7801 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7861 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7921 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7981 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     8041 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     8101 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     8161 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     8221 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     8281 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     8341 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     8401 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     8461 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     8521 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     8581 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     8641 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     8701 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     8761 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8821 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8881 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8941 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     9001 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     9061 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact
     9121 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     9181 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     9241 gacaattgca tgaagaatct gcttagg
//
