LOCUS       sequence-328302-        3573 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pDONR221 encoding ubiquitin-tagged NLRP1-CT to generate the native 
            S1213 N-terminus (no stop).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3573)
  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 3573)
  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 S1213
FEATURES             Location/Qualifiers
     source          1..3573
                     /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(1698..1797)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1814..1833)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1815..1833)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1838..1854)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1838..1854)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1956..1975)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1967..2776
                     /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(2034..2053)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2923..3511
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3412..3431
                     /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 ggttccccct tgggagtcct cctgaaaatg atccataatg ccctgcgctt
      961 cattcccgtc acctctgtgg tgttgcttta ccaccgcgtc catcctgagg aagtcacctt
     1021 ccacctctac ctgatcccaa gtgactgctc cattcggaag gccatagatg atctagaaat
     1081 gaaattccag tttgtgcgaa tccacaagcc acccccgctg accccacttt atatgggctg
     1141 tcgttacact gtgtctgggt ctggttcagg gatgctggaa atactcccca aggaactgga
     1201 gctctgctat cgaagccctg gagaagacca gctgttctcg gagttctacg ttggccactt
     1261 gggatcaggg atcaggctgc aagtgaaaga caagaaagat gagactctgg tgtgggaggc
     1321 cttggtgaaa ccaggagatc tcatgcctgc aactactctg atccctccag cccgcatagc
     1381 cgtaccttca cctctggatg ccccgcagtt gctgcacttt gtggaccagt atcgagagca
     1441 gctgatagcc cgagtgacat cggtggaggt tgtcttggac aaactgcatg gacaggtgct
     1501 gagccaggag cagtacgaga gggtgctggc tgagaacacg aggcccagcc agatgcggaa
     1561 gctgttcagc ttgagccagt cctgggaccg gaagtgcaaa gatggactct accaagccct
     1621 gaaggagacc catcctcacc tcattatgga actctgggag aagggcagca aaaagggact
     1681 cctgccactc agcagcgacc cagctttctt gtacaaagtt ggcattataa gaaagcattg
     1741 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     1801 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     1861 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     1921 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
     1981 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
     2041 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
     2101 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     2161 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     2221 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
     2281 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     2341 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     2401 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     2461 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
     2521 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     2581 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     2641 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     2701 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     2761 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     2821 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     2881 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2941 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3001 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3061 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3121 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3181 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3241 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3301 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3361 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3421 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3481 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3541 cctggccttt tgctggcctt ttgctcacat gtt
//
