LOCUS       sequence-331189-        4706 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry plasmid with 3xflag-pyrin (Wild-type).
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-WT-MEFV
FEATURES             Location/Qualifiers
     source          1..4706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      119..205
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      297..324
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     332..355
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    374..473
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             516..581
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(2951..3050)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3162..3181)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3173..3982
                     /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(3240..3259)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4072..4660
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4561..4580
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
       61 ctagcatgga tctcggggac gtctaactac taagcgagag tagggaactg ccaggcatca
      121 aataaaacga aaggctcagt cggaagactg ggcctttcgt tttatctgtt gtttgtcggt
      181 gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg tgaagcaacg
      241 gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ctaagcagaa
      301 ggccatcctg acggatggcc tttttgcgtt tctacaaact cttcctgtta gttagttact
      361 taagctcggg ccccaaataa tgattttatt ttgactgata gtgacctgtt cgttgcaaca
      421 aattgataag caatgctttt ttataatgcc aactttgtac aaaaaagcag gctttaaagg
      481 aaccaattca gtcgactgga tccggtacca ccatggacta caaagaccat gacggtgatt
      541 ataaagatca tgacatcgat tacaaggatg acgatgacaa gcttgcggcc gcggctaaga
      601 cccctagtga ccatctgctg tccaccctgg aggagctggt gccctatgac ttcgagaagt
      661 tcaagttcaa gctgcagaac accagtgtgc agaaggagca ctccaggatc ccccggagcc
      721 agatccagag agccaggccg gtgaagatgg ccactctgct ggtcacctac tatggggaag
      781 agtacgccgt gcagctcacc ctgcaggtcc tgcgggccat caaccagcgc ctgctggccg
      841 aggagctcca cagggcagcc attcaggaat attccacaca agaaaacggc acagatgatt
      901 ccgcagcgtc cagctccctg ggggagaaca agcccaggag cctgaagact ccagaccacc
      961 ccgaggggaa cgaggggaac ggccctcggc cgtacggggg cggagctgcc agcctgcggt
     1021 gcagccagcc cgaggccggg agggggctgt cgaggaagcc cctgagcaaa cgcagagaga
     1081 aggcctcgga gggcctggac gcgcagggca agcctcggac ccggagcccg gccctgccgg
     1141 gcgggagaag ccccggcccc tgcagggcgc tagagggggg ccaggccgag gtccggctgc
     1201 gcagaaacgc cagctccgcg gggaggctgc aggggctggc ggggggcgcc ccggggcaga
     1261 aggagtgcag gcccttcgaa gtgtacctgc cctcgggaaa gatgcgacct agaagccttg
     1321 aggtcaccat ttctacaggg gagaaggcgc ccgcaaatcc agaaattctc ctgactctag
     1381 aggaaaagac agctgcgaat ctggactcgg caacagaacc ccgggcaagg cccactccgg
     1441 atggaggggc atctgcggac ctgaaggaag gccctggaaa tccagaacat tcggtcaccg
     1501 gaaggccacc agacacggct gcgagtcccc gctgccacgc ccaggaagga gacccagttg
     1561 acggtacctg tgtgcgtgat tcctgcagct tccccgaggc agtttctggg cacccccagg
     1621 cctcaggcag ccgctcacct ggctgccccc ggtgccagga ctcccatgaa aggaagagcc
     1681 cgggaagcct aagcccccag cccctgccac agtgtaagcg ccacctgaag caggtccagc
     1741 tgctcttctg tgaggatcac gatgagccca tctgcctcat ctgcagtctg agtcaggagc
     1801 accaaggcca ccgggtgcgc cccattgagg aggtcgccct ggaacacaag aagaaaattc
     1861 agaagcagct ggagcatctg aagaagctga gaaaatcagg ggaggagcag cgatcctatg
     1921 gggaggagaa ggcagtgagc tttctgaaac aaactgaagc gctgaagcag cgggtgcaga
     1981 ggaagctgga gcaggtgtac tacttcctgg agcagcaaga gcatttcttt gtggcctcac
     2041 tggaggacgt gggccagatg gttgggcaga tcaggaaggc atatgacacc cgcgtatccc
     2101 aggacatcgc cctgctcgat gcgctgattg gggaactgga ggccaaggag tgccagtcag
     2161 aatgggaact tctgcaggac attggagaca tcttgcacag ggctaagaca gtgcctgtcc
     2221 ctgaaaagtg gaccactcct caagagataa aacaaaagat ccaactcctc caccagaagt
     2281 cagagtttgt ggagaagagc acaaagtact tctcagaaac cctgcgttca gaaatggaaa
     2341 tgttcaatgt tccagagctg attggcgctc aggcacatgc tgttaatgtg attctggatg
     2401 cagaaaccgc ttaccccaac ctcatcttct ctgatgatct gaagagtgtt agacttggaa
     2461 acaagtggga gaggctgcct gatggcccgc aaagatttga cagctgtatc attgttctgg
     2521 gctctccgag tttcctctct ggccgccgtt actgggaggt ggaggttgga gacaagacag
     2581 catggatcct gggagcctgc aagacatcca taagcaggaa agggaacatg actctgtcgc
     2641 cagagaatgg ctactgggtg gtgataatga tgaaggaaaa tgagtaccag gcgtccagcg
     2701 ttcccccgac ccgcctgcta ataaaggagc ctcccaagcg tgtgggcatc ttcgtggact
     2761 acagagttgg aagcatctcc ttttacaatg tgacagccag atcccacatc tatacattcg
     2821 ccagctgctc tttctctggg ccccttcaac ctatcttcag ccctgggaca cgtgatggag
     2881 ggaagaacac agctcctctg actatctgtc cagtgggtgg tcaggggcct gactgactcg
     2941 agatatctag acccagcttt cttgtacaaa gttggcatta taagaaagca ttgcttatca
     3001 atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct
     3061 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     3121 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     3181 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     3241 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     3301 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     3361 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     3421 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     3481 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     3541 agtgtccctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     3601 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     3661 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     3721 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     3781 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     3841 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     3901 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     3961 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacatt attcagattg
     4021 ggccccgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4081 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4141 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4201 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4261 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4321 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4381 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4441 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4501 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4561 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4621 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4681 tttacggttc ctggcctttt gctggc
//
