LOCUS Exported 4706 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Gateway entry plasmid with 3xFlag-pyrin (S242R/PAAND variant). ACCESSION . VERSION . KEYWORDS pENTR1A-3xFlag-S242R-MEFV SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4706) TITLE Functional assessment of disease associated polymorphisms in pyrin JOURNAL Unpublished REFERENCE 2 (bases 1 to 4706) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4706 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 46..73 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" primer_bind 81..104 /label=pENTR-F /note="pENTR vectors, forward primer" protein_bind 123..222 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 265..330 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" protein_bind complement(2700..2799) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(2911..2930) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 2922..3731 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVSLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(2989..3008) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 3821..4409 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4310..4329 /label=pBR322ori-F /note="pBR322 origin, forward primer" terminator 4574..4660 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" ORIGIN 1 ggcgggcagg acgcccgcca taaactgcca ggcatcaaac taagcagaag gccatcctga 61 cggatggcct ttttgcgttt ctacaaactc ttcctgttag ttagttactt aagctcgggc 121 cccaaataat gattttattt tgactgatag tgacctgttc gttgcaacaa attgataagc 181 aatgcttttt tataatgcca actttgtaca aaaaagcagg ctttaaagga accaattcag 241 tcgactggat ccggtaccac catggactac aaagaccatg acggtgatta taaagatcat 301 gacatcgatt acaaggatga cgatgacaag cttgcggccg cggctaagac ccctagtgac 361 catctgctgt ccaccctgga ggagctggtg ccctatgact tcgagaagtt caagttcaag 421 ctgcagaaca ccagtgtgca gaaggagcac tccaggatcc cccggagcca gatccagaga 481 gccaggccgg tgaagatggc cactctgctg gtcacctact atggggaaga gtacgccgtg 541 cagctcaccc tgcaggtcct gcgggccatc aaccagcgcc tgctggccga ggagctccac 601 agggcagcca ttcaggaata ttccacacaa gaaaacggca cagatgattc cgcagcgtcc 661 agctccctgg gggagaacaa gcccaggagc ctgaagactc cagaccaccc cgaggggaac 721 gaggggaacg gccctcggcc gtacgggggc ggagctgcca gcctgcggtg cagccagccc 781 gaggccggga gggggctgtc gaggaagccc ctgagcaaac gcagagagaa ggcctcggag 841 ggcctggacg cgcagggcaa gcctcggacc cggagcccgg ccctgccggg cgggagaagc 901 cccggcccct gcagggcgct agaggggggc caggccgagg tccggctgcg cagaaacgcc 961 agctccgcgg ggaggctgca ggggctggcg gggggcgccc cggggcagaa ggagtgcagg 1021 cccttcgaag tgtacctgcc ctcgggaaag atgcgaccta gaagacttga ggtcaccatt 1081 tctacagggg agaaggcgcc cgcaaatcca gaaattctcc tgactctaga ggaaaagaca 1141 gctgcgaatc tggactcggc aacagaaccc cgggcaaggc ccactccgga tggaggggca 1201 tctgcggacc tgaaggaagg ccctggaaat ccagaacatt cggtcaccgg aaggccacca 1261 gacacggctg cgagtccccg ctgccacgcc caggaaggag acccagttga cggtacctgt 1321 gtgcgtgatt cctgcagctt ccccgaggca gtttctgggc acccccaggc ctcaggcagc 1381 cgctcacctg gctgcccccg gtgccaggac tcccatgaaa ggaagagccc gggaagccta 1441 agcccccagc ccctgccaca gtgtaagcgc cacctgaagc aggtccagct gctcttctgt 1501 gaggatcacg atgagcccat ctgcctcatc tgcagtctga gtcaggagca ccaaggccac 1561 cgggtgcgcc ccattgagga ggtcgccctg gaacacaaga agaaaattca gaagcagctg 1621 gagcatctga agaagctgag aaaatcaggg gaggagcagc gatcctatgg ggaggagaag 1681 gcagtgagct ttctgaaaca aactgaagcg ctgaagcagc gggtgcagag gaagctggag 1741 caggtgtact acttcctgga gcagcaagag catttctttg tggcctcact ggaggacgtg 1801 ggccagatgg ttgggcagat caggaaggca tatgacaccc gcgtatccca ggacatcgcc 1861 ctgctcgatg cgctgattgg ggaactggag gccaaggagt gccagtcaga atgggaactt 1921 ctgcaggaca ttggagacat cttgcacagg gctaagacag tgcctgtccc tgaaaagtgg 1981 accactcctc aagagataaa acaaaagatc caactcctcc accagaagtc agagtttgtg 2041 gagaagagca caaagtactt ctcagaaacc ctgcgttcag aaatggaaat gttcaatgtt 2101 ccagagctga ttggcgctca ggcacatgct gttaatgtga ttctggatgc agaaaccgct 2161 taccccaacc tcatcttctc tgatgatctg aagagtgtta gacttggaaa caagtgggag 2221 aggctgcctg atggcccgca aagatttgac agctgtatca ttgttctggg ctctccgagt 2281 ttcctctctg gccgccgtta ctgggaggtg gaggttggag acaagacagc atggatcctg 2341 ggagcctgca agacatccat aagcaggaaa gggaacatga ctctgtcgcc agagaatggc 2401 tactgggtgg tgataatgat gaaggaaaat gagtaccagg cgtccagcgt tcccccgacc 2461 cgcctgctaa taaaggagcc tcccaagcgt gtgggcatct tcgtggacta cagagttgga 2521 agcatctcct tttacaatgt gacagccaga tcccacatct atacattcgc cagctgctct 2581 ttctctgggc cccttcaacc tatcttcagc cctgggacac gtgatggagg gaagaacaca 2641 gctcctctga ctatctgtcc agtgggtggt caggggcctg actgactcga gatatctaga 2701 cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa 2761 cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg cagctctggc 2821 ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa 2881 taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg 2941 aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg 3001 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg 3061 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga 3121 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc 3181 gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg 3241 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgtccctgc 3301 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc 3361 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg 3421 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct 3481 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg 3541 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc 3601 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc 3661 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg 3721 agtttttcta atcagaattg gttaattggt tgtaacatta ttcagattgg gccccgttcc 3781 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc 3841 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg 3901 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa 3961 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc 4021 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt 4081 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa 4141 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc 4201 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc 4261 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct 4321 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat 4381 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc 4441 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg 4501 ataaccgtat taccgctagc atggatctcg gggacgtcta actactaagc gagagtaggg 4561 aactgccagg catcaaataa aacgaaaggc tcagtcggaa gactgggcct ttcgttttat 4621 ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa 4681 cgttgtgaag caacggcccg gagggt //