LOCUS       sequence-335590-        3381 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pDONR221 for Gateway shuttling. Contains the CARD8 FIIND 
            (L162M-P434, no stop codon) with an interface III mutation (L368G).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3381)
  AUTHORS   Sharif H, Hollingsworth LR, Griswold AR, Hsiao JC, Wang Q, 
            Bachovchin DA, Wu H
  TITLE     Dipeptidyl peptidase 9 sets a threshold for CARD8 inflammasome 
            formation by sequestering its active C-terminal fragment.
  JOURNAL   Immunity. 2021 Jul 13;54(7):1392-1404.e10. doi: 
            10.1016/j.immuni.2021.04.024. Epub 2021 May 20.
  PUBMED    34019797
REFERENCE   2  (bases 1 to 3381)
  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"; doi: 
            "10.1016/j.immuni.2021.04.024"; journalName: "Immunity"; date: 
            "2021-07-13- 13"; volume: "54"; issue: "7"; pages: "1392-1404.e10"
COMMENT     Sequence Label: pDONR221 CARD8 FIIND L368G
FEATURES             Location/Qualifiers
     source          1..3381
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(126..714)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(206..225)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(861..1670)
                     /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     1584..1603
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1662..1681
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1783..1799
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1783..1799
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1804..1823
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1804..1822
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1840..1939
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2777..2876)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2892..2909)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2892..2908)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2901..2923)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      2972..3058
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3150..3177
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3336..3353)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
       61 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
      121 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
      181 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
      241 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
      301 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
      361 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
      421 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
      481 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
      541 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
      601 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
      661 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
      721 cctttgatct tttctacggg gtctgacgct cagtggaacg acgcgtaact cacgttaagg
      781 gattttggtc atgagcttgc gccgtcccgt caagtcagcg taatgctctg ccagtgttac
      841 aaccaattaa ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta
      901 ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
      961 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
     1021 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
     1081 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc
     1141 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa
     1201 ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga
     1261 caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata
     1321 ttttcacctg aatcaggata ttcttctaat acctggaatg ctgtttttcc ggggatcgca
     1381 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc
     1441 ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta
     1501 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa gcgatagatt
     1561 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
     1621 atgttggaat ttaatcgcgg cctcgacgtt tcccgttgaa tatggctcat aacacccctt
     1681 gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt
     1741 gcaatgtaac atcagagatt ttgagacacg ggccagagct gccaggaaac agctatgacc
     1801 atgtaatacg actcactata ggggatatca gctggatggc aaataatgat tttattttga
     1861 ctgatagtga cctgttcgtt gcaacaaatt gataagcaat gctttcttat aatgccaact
     1921 ttgtacaaga aagctgggtc tggcttcacc tcagtatccc acaccaaagt cccatgtctt
     1981 ttttcagtaa tctcaagttg aatgggttcc ttcatctgcc cagcatagaa ttttgagaag
     2041 tgctgaattt ctccagggct cctgtaggac aatttcaact ccttgggcat tactttcagg
     2101 ttagcagaat tagacacaat ataactggaa ccaaagttcc cgggttccat tgggggcgaa
     2161 gtctgcaggc gcacaccatg gaagcgatct tcctcatcat ctatcgcctt tgttagcaag
     2221 gcgtcgctgg ggacaaggta caagtggaac ttaatatctt cggggtgggg gtgataatag
     2281 atcaatgtgt tggaagtgat ggggatggag aggcgagtcc cactggcgat ccgcagcagg
     2341 atgcccatca gagagaagct ggggctttcc aggacagcat agaaaggctc cacccgggct
     2401 ggatgctcca ggaccatccc ttcattctta aaatgggcaa cgagaaacca ggagacgtcc
     2461 acctcacctg cttggaggga gatgaagtgg gggaggtgga tttcggcgac agcctcctct
     2521 ggctctgcag tgacatcaaa caaggggccg cccaccagcc actgttcatg gtgctgcagg
     2581 tccagggcca ggtgctgact ccaggaacca aacgcaatcg tcactgtgac ctcatccctt
     2641 accaggaagc cgaggcctgt ggctgaccac agataccagc cagcagtggg gaaccaaacg
     2701 ctgtatctgt ttgtgctctt atcaatcaac tcaacatcca catttccttc aggccccatg
     2761 gttctatctc cttcgaagcc tgcttttttg tacaaagttg gcattataaa aaagcattgc
     2821 tcatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttggggc
     2881 ccgagcttaa gactggccgt cgttttacaa cgtcgtgact gggaaaacat ccatgctagc
     2941 gttaacgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcggaaga
     3001 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     3061 gccgggagcg gatttgaacg ttgtgaagca acggcccgga gggtggcggg caggacgccc
     3121 gccataaact gccaggcatc aaactaagca gaaggccatc ctgacggatg gcctttttgc
     3181 gtttctacaa actcttcctg gctagcggta cgcgtattaa ttgcgttgcg ctcactgccc
     3241 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3301 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3361 gtcgttcggc tgcggcgagc g
//
