LOCUS       sequence-328303-        3567 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pDONR221 encoding ubiquitin-tagged NLRP1-CT to generate a L1215 
            N-terminus (no stop).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3567)
  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 3567)
  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 L1215
FEATURES             Location/Qualifiers
     source          1..3567
                     /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(1692..1791)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1808..1827)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1809..1827)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1832..1848)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1832..1848)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1950..1969)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1961..2770
                     /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(2028..2047)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2917..3505
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3406..3425
                     /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 ggtttgggag tcctcctgaa aatgatccat aatgccctgc gcttcattcc
      961 cgtcacctct gtggtgttgc tttaccaccg cgtccatcct gaggaagtca ccttccacct
     1021 ctacctgatc ccaagtgact gctccattcg gaaggccata gatgatctag aaatgaaatt
     1081 ccagtttgtg cgaatccaca agccaccccc gctgacccca ctttatatgg gctgtcgtta
     1141 cactgtgtct gggtctggtt cagggatgct ggaaatactc cccaaggaac tggagctctg
     1201 ctatcgaagc cctggagaag accagctgtt ctcggagttc tacgttggcc acttgggatc
     1261 agggatcagg ctgcaagtga aagacaagaa agatgagact ctggtgtggg aggccttggt
     1321 gaaaccagga gatctcatgc ctgcaactac tctgatccct ccagcccgca tagccgtacc
     1381 ttcacctctg gatgccccgc agttgctgca ctttgtggac cagtatcgag agcagctgat
     1441 agcccgagtg acatcggtgg aggttgtctt ggacaaactg catggacagg tgctgagcca
     1501 ggagcagtac gagagggtgc tggctgagaa cacgaggccc agccagatgc ggaagctgtt
     1561 cagcttgagc cagtcctggg accggaagtg caaagatgga ctctaccaag ccctgaagga
     1621 gacccatcct cacctcatta tggaactctg ggagaagggc agcaaaaagg gactcctgcc
     1681 actcagcagc gacccagctt tcttgtacaa agttggcatt ataagaaagc attgcttatc
     1741 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
     1801 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
     1861 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     1921 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     1981 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     2041 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc
     2101 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
     2161 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
     2221 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt
     2281 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
     2341 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
     2401 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
     2461 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
     2521 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
     2581 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
     2641 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
     2701 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
     2761 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg
     2821 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg
     2881 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2941 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3001 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3061 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3121 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3181 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3241 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3301 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3361 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3421 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3481 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3541 cttttgctgg ccttttgctc acatgtt
//
