LOCUS       sequence-328298-        6972 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Gateway pDONR vector encoding NLRP1 with the patient mutation T755N 
            (Drutman et al., 2019) (no stop codon).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6972)
  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 6972)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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 T755N
FEATURES             Location/Qualifiers
     source          1..6972
                     /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(5097..5196)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5213..5232)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5214..5232)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5237..5253)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5237..5253)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5355..5374)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5366..6175
                     /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(5433..5452)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      6322..6910
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6811..6830
                     /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 tagtgtgcaa
     2941 tttctgcatt aaattcagcc gccacgtgaa gaagcttcag ctgattgagg gcaggcagca
     3001 cagatcaaca tggagcccca ccatggtagt cctgttcagg tgggtcccag tcacagatgc
     3061 ctattggcag attctcttct ccgtcctcaa ggtcaccaga aacctgaagg agctggacct
     3121 aagtggaaac tcgctgagcc actctgcagt gaagagtctt tgtaagaccc tgagacgccc
     3181 tcgctgcctc ctggagaccc tgcggttggc tggctgtggc ctcacagctg aggactgcaa
     3241 ggaccttgcc tttgggctga gagccaacca gaccctgacc gagctggacc tgagcttcaa
     3301 tgtgctcacg gatgctggag ccaaacacct ttgccagaga ctgagacagc cgagctgcaa
     3361 gctacagcga ctgcagctgg tcagctgtgg cctcacgtct gactgctgcc aggacctggc
     3421 ctctgtgctt agtgccagcc ccagcctgaa ggagctagac ctgcagcaga acaacctgga
     3481 tgacgttggc gtgcgactgc tctgtgaggg gctcaggcat cctgcctgca aactcatacg
     3541 cctggggctg gaccagacaa ctctgagtga tgagatgagg caggaactga gggccctgga
     3601 gcaggagaaa cctcagctgc tcatcttcag cagacggaaa ccaagtgtga tgacccctac
     3661 tgagggcctg gatacgggag agatgagtaa tagcacatcc tcactcaagc ggcagagact
     3721 cggatcagag agggcggctt cccatgttgc tcaggctaat ctcaaactcc tggacgtgag
     3781 caagatcttc ccaattgctg agattgcaga ggaaagctcc ccagaggtag taccggtgga
     3841 actcttgtgc gtgccttctc ctgcctctca aggggacctg catacgaagc ctttggggac
     3901 tgacgatgac ttctggggcc ccacggggcc tgtggctact gaggtagttg acaaagaaaa
     3961 gaacttgtac cgagttcact tccctgtagc tggctcctac cgctggccca acacgggtct
     4021 ctgctttgtg atgagagaag cggtgaccgt tgagattgaa ttctgtgtgt gggaccagtt
     4081 cctgggtgag atcaacccac agcacagctg gatggtggca gggcctctgc tggacatcaa
     4141 ggctgagcct ggagctgtgg aagctgtgca cctccctcac tttgtggctc tccaaggggg
     4201 ccatgtggac acatccctgt tccaaatggc ccactttaaa gaggagggga tgctcctgga
     4261 gaagccagcc agggtggagc tgcatcacat agttctggaa aaccccagct tctccccctt
     4321 gggagtcctc ctgaaaatga tccataatgc cctgcgcttc attcccgtca cctctgtggt
     4381 gttgctttac caccgcgtcc atcctgagga agtcaccttc cacctctacc tgatcccaag
     4441 tgactgctcc attcggaagg ccatagatga tctagaaatg aaattccagt ttgtgcgaat
     4501 ccacaagcca cccccgctga ccccacttta tatgggctgt cgttacactg tgtctgggtc
     4561 tggttcaggg atgctggaaa tactccccaa ggaactggag ctctgctatc gaagccctgg
     4621 agaagaccag ctgttctcgg agttctacgt tggccacttg ggatcaggga tcaggctgca
     4681 agtgaaagac aagaaagatg agactctggt gtgggaggcc ttggtgaaac caggagatct
     4741 catgcctgca actactctga tccctccagc ccgcatagcc gtaccttcac ctctggatgc
     4801 cccgcagttg ctgcactttg tggaccagta tcgagagcag ctgatagccc gagtgacatc
     4861 ggtggaggtt gtcttggaca aactgcatgg acaggtgctg agccaggagc agtacgagag
     4921 ggtgctggct gagaacacga ggcccagcca gatgcggaag ctgttcagct tgagccagtc
     4981 ctgggaccgg aagtgcaaag atggactcta ccaagccctg aaggagaccc atcctcacct
     5041 cattatggaa ctctgggaga agggcagcaa aaagggactc ctgccactca gcagcgaccc
     5101 agctttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga
     5161 acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag
     5221 tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct
     5281 ctgatgttac attgcacaag ataaaaatat atcatcatga acaataaaac tgtctgctta
     5341 cataaacagt aatacaaggg gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg
     5401 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     5461 gcaatcaggt gcgacaatct atcgcttgta tgggaagccc gatgcgccag agttgtttct
     5521 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     5581 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     5641 atggttactc accactgcga tccccggaaa aacagcattc caggtattag aagaatatcc
     5701 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     5761 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
     5821 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     5881 tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg attcagtcgt
     5941 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     6001 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     6061 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     6121 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga
     6181 attggttaat tggttgtaac actggcagag cattacgctg acttgacggg acggcgcaag
     6241 ctcatgacca aaatccctta acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt
     6301 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6361 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6421 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6481 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6541 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6601 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6661 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6721 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6781 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6841 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6901 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6961 tgctcacatg tt
//
