LOCUS       sequence-335589-        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 (F370G).
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 L370G
FEATURES             Location/Qualifiers
     source          1..3381
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(192..291)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(307..324)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(307..323)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(316..338)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      387..473
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      565..592
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(751..768)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(922..1510)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1002..1021)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1657..2466)
                     /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     2380..2399
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2458..2477
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     2579..2595
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2579..2595
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2600..2619
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2600..2618
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2636..2735
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ggccaggtgc tgactccagg aaccaaacgc aatcgtcact gtgacctcat cccttaccag
       61 gaagccgagg cctgtggctg accacagata ccagccagca gtggggaacc aaacgctgta
      121 tctgtttgtg ctcttatcaa tcaactcaac atccacattt ccttcaggcc ccatggttct
      181 atctccttcg aagcctgctt ttttgtacaa agttggcatt ataaaaaagc attgctcatc
      241 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt ggggcccgag
      301 cttaagactg gccgtcgttt tacaacgtcg tgactgggaa aacatccatg ctagcgttaa
      361 cgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg gaagactggg
      421 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg
      481 gagcggattt gaacgttgtg aagcaacggc ccggagggtg gcgggcagga cgcccgccat
      541 aaactgccag gcatcaaact aagcagaagg ccatcctgac ggatggcctt tttgcgtttc
      601 tacaaactct tcctggctag cggtacgcgt attaattgcg ttgcgctcac tgcccgcttt
      661 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
      721 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
      781 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
      841 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
      901 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
      961 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     1021 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     1081 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     1141 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     1201 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     1261 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     1321 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     1381 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     1441 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     1501 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgacgc
     1561 gtaactcacg ttaagggatt ttggtcatga gcttgcgccg tcccgtcaag tcagcgtaat
     1621 gctctgccag tgttacaacc aattaaccaa ttctgattag aaaaactcat cgagcatcaa
     1681 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     1741 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     1801 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     1861 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     1921 tttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     1981 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg
     2041 atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     2101 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     2161 ttttccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     2221 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     2281 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     2341 atacaagcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     2401 atataaatca gcatccatgt tggaatttaa tcgcggcctc gacgtttccc gttgaatatg
     2461 gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga
     2521 tatattttta tcttgtgcaa tgtaacatca gagattttga gacacgggcc agagctgcca
     2581 ggaaacagct atgaccatgt aatacgactc actatagggg atatcagctg gatggcaaat
     2641 aatgatttta ttttgactga tagtgacctg ttcgttgcaa caaattgata agcaatgctt
     2701 tcttataatg ccaactttgt acaagaaagc tgggtctggc ttcacctcag tatcccacac
     2761 caaagtccca tgtctttttt cagtaatctc aagttgaatg ggttccttca tctgcccagc
     2821 atagaatttt gagaagtgct gaatttctcc agggctcctg taggacaatt tcaactcctt
     2881 gggcattact ttcaggttag cagaattaga cacaatataa ctggaaccac cgttcagggg
     2941 ttccattggg ggcgaagtct gcaggcgcac accatggaag cgatcttcct catcatctat
     3001 cgcctttgtt agcaaggcgt cgctggggac aaggtacaag tggaacttaa tatcttcggg
     3061 gtgggggtga taatagatca atgtgttgga agtgatgggg atggagaggc gagtcccact
     3121 ggcgatccgc agcaggatgc ccatcagaga gaagctgggg ctttccagga cagcatagaa
     3181 aggctccacc cgggctggat gctccaggac catcccttca ttcttaaaat gggcaacgag
     3241 aaaccaggag acgtccacct cacctgcttg gagggagatg aagtggggga ggtggatttc
     3301 ggcgacagcc tcctctggct ctgcagtgac atcaaacaag gggccgccca ccagccactg
     3361 ttcatggtgc tgcaggtcca g
//
