LOCUS       sequence-328305-        6801 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway pDONR vector encoding NLRP1 with the patient mutation 
            _787-843 (Zhong et al., 2016) (no stop codon).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6801)
  AUTHORS   Hollingsworth LR, Sharif H, Griswold AR, Fontana P, Mintseris J, 
            Dagbay KB, Paulo JA, Gygi SP, Bachovchin DA, Wu H
  TITLE     DPP9 sequesters the C terminus of NLRP1 to repress inflammasome 
            activation.
  JOURNAL   Nature. 2021 Mar 17. pii: 10.1038/s41586-021-03350-4. doi: 
            10.1038/s41586-021-03350-4.
  PUBMED    33731932
REFERENCE   2  (bases 1 to 6801)
  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: "Nature. 
            2021 Mar 17. pii: 10.1038/s41586-021-03350-4. doi: 
            10.1038/s41586-021-03350-4."
COMMENT     Sequence Label: pDONR221 NLRP1 delta787-843
FEATURES             Location/Qualifiers
     source          1..6801
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      671..680
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2984..2995
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(4926..5025)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5042..5061)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5043..5061)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5066..5082)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5066..5082)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5184..5203)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5195..6004
                     /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
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(5262..5281)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      6151..6739
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6640..6659
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggcttc gccaccatgg ctggcggagc ctggggccgc ctggcctgtt acttggagtt
      721 cctgaagaag gaggagctga aggagttcca gcttctgctc gccaataaag cgcactccag
      781 gagctcttcg ggtgagacac ccgctcagcc agagaagacg agtggcatgg aggtggcctc
      841 gtacctggtg gctcagtatg gggagcagcg ggcctgggac ctagccctcc atacctggga
      901 gcagatgggg ctgaggtcac tgtgcgccca agcccaggaa ggggcaggcc actctccctc
      961 attcccctac agcccaagtg aaccccacct ggggtctccc agccaaccca cctccaccgc
     1021 agtgctaatg ccctggatcc atgaattgcc ggcggggtgc acccagggct cagagagaag
     1081 ggttttgaga cagctgcctg acacatctgg acgccgctgg agagaaatct ctgcctcact
     1141 cctctaccaa gctcttccaa gctccccaga ccatgagtct ccaagccagg agtcacccaa
     1201 cgcccccaca tccacagcag tgctggggag ctggggatcc ccacctcagc ccagcctagc
     1261 acccagagag caggaggctc ctgggaccca atggcctctg gatgaaacgt caggaattta
     1321 ctacacagaa atcagagaaa gagagagaga gaaatcagag aaaggcaggc ccccatgggc
     1381 agcggtggta ggaacgcccc cacaggcgca caccagccta cagccccacc accacccatg
     1441 ggagccttct gtgagagaga gcctctgttc cacatggccc tggaaaaatg aggattttaa
     1501 ccaaaaattc acacagctgc tacttctaca aagacctcac cccagaagcc aagatcccct
     1561 ggtcaagaga agctggcctg attatgtgga ggagaatcga ggacatttaa ttgagatcag
     1621 agacttattt ggcccaggcc tggataccca agaacctcgc atagtcatac tgcagggggc
     1681 tgctggaatt gggaagtcaa cactggccag gcaggtgaag gaagcctggg ggagaggcca
     1741 gctgtatggg gaccgcttcc agcatgtctt ctacttcagc tgcagagagc tggcccagtc
     1801 caaggtggtg agtctcgctg agctcatcgg aaaagatggg acagccactc cggctcccat
     1861 tagacagatc ctgtctaggc cagagcggct gctcttcatc ctcgatggtg tagatgagcc
     1921 aggatgggtc ttgcaggagc cgagttctga gctctgtctg cactggagcc agccacagcc
     1981 ggcggatgca ctgctgggca gtttgctggg gaaaactata cttcccgagg catccttcct
     2041 gatcacggct cggaccacag ctctgcagaa cctcattcct tctttggagc aggcacgttg
     2101 ggtagaggtc ctggggttct ctgagtccag caggaaggaa tatttctaca gatatttcac
     2161 agatgaaagg caagcaatta gagcctttag gttggtcaaa tcaaacaaag agctctgggc
     2221 cctgtgtctt gtgccctggg tgtcctggct ggcctgcact tgcctgatgc agcagatgaa
     2281 gcggaaggaa aaactcacac tgacttccaa gaccaccaca accctctgtc tacattacct
     2341 tgcccaggct ctccaagctc agccattggg accccagctc agagacctct gctctctggc
     2401 tgctgagggc atctggcaaa aaaagaccct tttcagtcca gatgacctca ggaagcatgg
     2461 gttagatggg gccatcatct ccaccttctt gaagatgggt attcttcaag agcaccccat
     2521 ccctctgagc tacagcttca ttcacctctg tttccaagag ttctttgcag caatgtccta
     2581 tgtcttggag gatgagaagg ggagaggtaa acattctaat tgcatcatag atttggaaaa
     2641 gacgctagaa gcatatggaa tacatggcct gtttggggca tcaaccacac gtttcctatt
     2701 gggcctgtta agtgatgagg gggagagaga gatggagaac atctttcact gccggctgtc
     2761 tcaggggagg aacctgatgc agtgggtccc gtccctgcag ctgctgctgc agccacactc
     2821 tctggagtcc ctccactgct tgtacgagac tcggaacaaa acgttcctga cacaagtgat
     2881 ggcccatttc gaagaaatgg gcatgtgtgt agaaacagac atggagctct tagtgtgcac
     2941 tttctgcatt aaattcagcc gccacgtgaa gaagcttcag ctgattgagg gcaggcagca
     3001 cagatcaaca tggagcccca ccatggtagt cctgttggct ggctgtggcc tcacagctga
     3061 ggactgcaag gaccttgcct ttgggctgag agccaaccag accctgaccg agctggacct
     3121 gagcttcaat gtgctcacgg atgctggagc caaacacctt tgccagagac tgagacagcc
     3181 gagctgcaag ctacagcgac tgcagctggt cagctgtggc ctcacgtctg actgctgcca
     3241 ggacctggcc tctgtgctta gtgccagccc cagcctgaag gagctagacc tgcagcagaa
     3301 caacctggat gacgttggcg tgcgactgct ctgtgagggg ctcaggcatc ctgcctgcaa
     3361 actcatacgc ctggggctgg accagacaac tctgagtgat gagatgaggc aggaactgag
     3421 ggccctggag caggagaaac ctcagctgct catcttcagc agacggaaac caagtgtgat
     3481 gacccctact gagggcctgg atacgggaga gatgagtaat agcacatcct cactcaagcg
     3541 gcagagactc ggatcagaga gggcggcttc ccatgttgct caggctaatc tcaaactcct
     3601 ggacgtgagc aagatcttcc caattgctga gattgcagag gaaagctccc cagaggtagt
     3661 accggtggaa ctcttgtgcg tgccttctcc tgcctctcaa ggggacctgc atacgaagcc
     3721 tttggggact gacgatgact tctggggccc cacggggcct gtggctactg aggtagttga
     3781 caaagaaaag aacttgtacc gagttcactt ccctgtagct ggctcctacc gctggcccaa
     3841 cacgggtctc tgctttgtga tgagagaagc ggtgaccgtt gagattgaat tctgtgtgtg
     3901 ggaccagttc ctgggtgaga tcaacccaca gcacagctgg atggtggcag ggcctctgct
     3961 ggacatcaag gctgagcctg gagctgtgga agctgtgcac ctccctcact ttgtggctct
     4021 ccaagggggc catgtggaca catccctgtt ccaaatggcc cactttaaag aggaggggat
     4081 gctcctggag aagccagcca gggtggagct gcatcacata gttctggaaa accccagctt
     4141 ctcccccttg ggagtcctcc tgaaaatgat ccataatgcc ctgcgcttca ttcccgtcac
     4201 ctctgtggtg ttgctttacc accgcgtcca tcctgaggaa gtcaccttcc acctctacct
     4261 gatcccaagt gactgctcca ttcggaaggc catagatgat ctagaaatga aattccagtt
     4321 tgtgcgaatc cacaagccac ccccgctgac cccactttat atgggctgtc gttacactgt
     4381 gtctgggtct ggttcaggga tgctggaaat actccccaag gaactggagc tctgctatcg
     4441 aagccctgga gaagaccagc tgttctcgga gttctacgtt ggccacttgg gatcagggat
     4501 caggctgcaa gtgaaagaca agaaagatga gactctggtg tgggaggcct tggtgaaacc
     4561 aggagatctc atgcctgcaa ctactctgat ccctccagcc cgcatagccg taccttcacc
     4621 tctggatgcc ccgcagttgc tgcactttgt ggaccagtat cgagagcagc tgatagcccg
     4681 agtgacatcg gtggaggttg tcttggacaa actgcatgga caggtgctga gccaggagca
     4741 gtacgagagg gtgctggctg agaacacgag gcccagccag atgcggaagc tgttcagctt
     4801 gagccagtcc tgggaccgga agtgcaaaga tggactctac caagccctga aggagaccca
     4861 tcctcacctc attatggaac tctgggagaa gggcagcaaa aagggactcc tgccactcag
     4921 cagcgaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     4981 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     5041 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     5101 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     5161 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     5221 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     5281 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     5341 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     5401 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     5461 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     5521 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     5581 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     5641 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     5701 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     5761 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     5821 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     5881 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     5941 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     6001 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     6061 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     6121 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6181 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6241 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     6301 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     6361 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6421 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6481 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6541 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6601 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6661 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6721 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6781 ctggcctttt gctcacatgt t
//
