LOCUS       sequence-328291-        3558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pDONR221 encoding ubiquitin-tagged NLRP1-CT to generate a L1218 
            N-terminus (no stop).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3558)
  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 3558)
  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 Ub-NLRP1 L1218
FEATURES             Location/Qualifiers
     source          1..3558
                     /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"
     CDS             686..913
                     /codon_start=1
                     /product="human ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     protein_bind    complement(1683..1782)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1799..1818)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1800..1818)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1823..1839)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1823..1839)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1941..1960)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1952..2761
                     /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(2019..2038)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2908..3496
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3397..3416
                     /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 gaaggagata gaaccatgca gatcttcgtc aagacgttaa ccggtaaaac
      721 cataactctg gaagttgaac catccgatac catcgaaaac gttaaggcta aaattcaaga
      781 caaggaagga attccacctg atcaacaaag attgatcttt gccggtaagc agctcgagga
      841 cggtagaacg ctgtctgatt acaacattca gaaggagtcg accttacatc ttgtcttaag
      901 actaagaggt ggtctcctga aaatgatcca taatgccctg cgcttcattc ccgtcacctc
      961 tgtggtgttg ctttaccacc gcgtccatcc tgaggaagtc accttccacc tctacctgat
     1021 cccaagtgac tgctccattc ggaaggccat agatgatcta gaaatgaaat tccagtttgt
     1081 gcgaatccac aagccacccc cgctgacccc actttatatg ggctgtcgtt acactgtgtc
     1141 tgggtctggt tcagggatgc tggaaatact ccccaaggaa ctggagctct gctatcgaag
     1201 ccctggagaa gaccagctgt tctcggagtt ctacgttggc cacttgggat cagggatcag
     1261 gctgcaagtg aaagacaaga aagatgagac tctggtgtgg gaggccttgg tgaaaccagg
     1321 agatctcatg cctgcaacta ctctgatccc tccagcccgc atagccgtac cttcacctct
     1381 ggatgccccg cagttgctgc actttgtgga ccagtatcga gagcagctga tagcccgagt
     1441 gacatcggtg gaggttgtct tggacaaact gcatggacag gtgctgagcc aggagcagta
     1501 cgagagggtg ctggctgaga acacgaggcc cagccagatg cggaagctgt tcagcttgag
     1561 ccagtcctgg gaccggaagt gcaaagatgg actctaccaa gccctgaagg agacccatcc
     1621 tcacctcatt atggaactct gggagaaggg cagcaaaaag ggactcctgc cactcagcag
     1681 cgacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg
     1741 caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc
     1801 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca
     1861 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc
     1921 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag
     1981 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     2041 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt
     2101 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     2161 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     2221 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga
     2281 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     2341 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     2401 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     2461 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
     2521 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     2581 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     2641 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     2701 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     2761 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg
     2821 cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
     2881 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2941 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3001 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3061 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3121 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3181 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3241 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3301 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3361 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3421 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3481 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3541 gccttttgct cacatgtt
//
