LOCUS       sequence-331191-        4706 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry plasmid with 3xFlag-pyrin (M694V/FMF variant).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4706)
  AUTHORS   Chirita D, Jamilloux Y, Henry T, Magnotti F
  TITLE     Functional Assessment of Disease-Associated Pyrin Variants.
  JOURNAL   Methods Mol Biol. 2022;2523:179-195. doi: 
            10.1007/978-1-0716-2449-4_12.
  PUBMED    35759198
REFERENCE   2  (bases 1 to 4706)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Methods Mol
            Biol."; date: "2022"; volume: "2523:179-195. doi"; pages: " 
            10.1007/978-1-0716-2449-4_12"
COMMENT     Sequence Label: pENTR1A-3xFlag-M694V-MEFV
FEATURES             Location/Qualifiers
     source          1..4706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      116..202
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      294..321
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     329..352
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    371..470
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             513..578
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(2948..3047)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3159..3178)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3170..3979
                     /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(3237..3256)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4069..4657
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4558..4577
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcta
       61 gcatggatct cggggacgtc taactactaa gcgagagtag ggaactgcca ggcatcaaat
      121 aaaacgaaag gctcagtcgg aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
      181 cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgtga agcaacggcc
      241 cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaacta agcagaaggc
      301 catcctgacg gatggccttt ttgcgtttct acaaactctt cctgttagtt agttacttaa
      361 gctcgggccc caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat
      421 tgataagcaa tgctttttta taatgccaac tttgtacaaa aaagcaggct ttaaaggaac
      481 caattcagtc gactggatcc ggtaccacca tggactacaa agaccatgac ggtgattata
      541 aagatcatga catcgattac aaggatgacg atgacaagct tgcggccgcg gctaagaccc
      601 ctagtgacca tctgctgtcc accctggagg agctggtgcc ctatgacttc gagaagttca
      661 agttcaagct gcagaacacc agtgtgcaga aggagcactc caggatcccc cggagccaga
      721 tccagagagc caggccggtg aagatggcca ctctgctggt cacctactat ggggaagagt
      781 acgccgtgca gctcaccctg caggtcctgc gggccatcaa ccagcgcctg ctggccgagg
      841 agctccacag ggcagccatt caggaatatt ccacacaaga aaacggcaca gatgattccg
      901 cagcgtccag ctccctgggg gagaacaagc ccaggagcct gaagactcca gaccaccccg
      961 aggggaacga ggggaacggc cctcggccgt acgggggcgg agctgccagc ctgcggtgca
     1021 gccagcccga ggccgggagg gggctgtcga ggaagcccct gagcaaacgc agagagaagg
     1081 cctcggaggg cctggacgcg cagggcaagc ctcggacccg gagcccggcc ctgccgggcg
     1141 ggagaagccc cggcccctgc agggcgctag aggggggcca ggccgaggtc cggctgcgca
     1201 gaaacgccag ctccgcgggg aggctgcagg ggctggcggg gggcgccccg gggcagaagg
     1261 agtgcaggcc cttcgaagtg tacctgccct cgggaaagat gcgacctaga agccttgagg
     1321 tcaccatttc tacaggggag aaggcgcccg caaatccaga aattctcctg actctagagg
     1381 aaaagacagc tgcgaatctg gactcggcaa cagaaccccg ggcaaggccc actccggatg
     1441 gaggggcatc tgcggacctg aaggaaggcc ctggaaatcc agaacattcg gtcaccggaa
     1501 ggccaccaga cacggctgcg agtccccgct gccacgccca ggaaggagac ccagttgacg
     1561 gtacctgtgt gcgtgattcc tgcagcttcc ccgaggcagt ttctgggcac ccccaggcct
     1621 caggcagccg ctcacctggc tgcccccggt gccaggactc ccatgaaagg aagagcccgg
     1681 gaagcctaag cccccagccc ctgccacagt gtaagcgcca cctgaagcag gtccagctgc
     1741 tcttctgtga ggatcacgat gagcccatct gcctcatctg cagtctgagt caggagcacc
     1801 aaggccaccg ggtgcgcccc attgaggagg tcgccctgga acacaagaag aaaattcaga
     1861 agcagctgga gcatctgaag aagctgagaa aatcagggga ggagcagcga tcctatgggg
     1921 aggagaaggc agtgagcttt ctgaaacaaa ctgaagcgct gaagcagcgg gtgcagagga
     1981 agctggagca ggtgtactac ttcctggagc agcaagagca tttctttgtg gcctcactgg
     2041 aggacgtggg ccagatggtt gggcagatca ggaaggcata tgacacccgc gtatcccagg
     2101 acatcgccct gctcgatgcg ctgattgggg aactggaggc caaggagtgc cagtcagaat
     2161 gggaacttct gcaggacatt ggagacatct tgcacagggc taagacagtg cctgtccctg
     2221 aaaagtggac cactcctcaa gagataaaac aaaagatcca actcctccac cagaagtcag
     2281 agtttgtgga gaagagcaca aagtacttct cagaaaccct gcgttcagaa atggaaatgt
     2341 tcaatgttcc agagctgatt ggcgctcagg cacatgctgt taatgtgatt ctggatgcag
     2401 aaaccgctta ccccaacctc atcttctctg atgatctgaa gagtgttaga cttggaaaca
     2461 agtgggagag gctgcctgat ggcccgcaaa gatttgacag ctgtatcatt gttctgggct
     2521 ctccgagttt cctctctggc cgccgttact gggaggtgga ggttggagac aagacagcat
     2581 ggatcctggg agcctgcaag acatccataa gcaggaaagg gaacatgact ctgtcgccag
     2641 agaatggcta ctgggtggtg ataatggtga aggaaaatga gtaccaggcg tccagcgttc
     2701 ccccgacccg cctgctaata aaggagcctc ccaagcgtgt gggcatcttc gtggactaca
     2761 gagttggaag catctccttt tacaatgtga cagccagatc ccacatctat acattcgcca
     2821 gctgctcttt ctctgggccc cttcaaccta tcttcagccc tgggacacgt gatggaggga
     2881 agaacacagc tcctctgact atctgtccag tgggtggtca ggggcctgac tgactcgaga
     2941 tatctagacc cagctttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt
     3001 tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgca
     3061 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     3121 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     3181 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     3241 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
     3301 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     3361 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     3421 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
     3481 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     3541 gtccctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     3601 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     3661 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
     3721 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     3781 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     3841 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     3901 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     3961 gctcgatgag tttttctaat cagaattggt taattggttg taacattatt cagattgggc
     4021 cccgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4081 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4141 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4201 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4261 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4321 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4381 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4441 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4501 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4561 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4621 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4681 acggttcctg gccttttgct ggcctt
//
