LOCUS       sequence-332697-        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 (F405G).
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 F405G
FEATURES             Location/Qualifiers
     source          1..3381
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     134..153
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     387..404
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(563..590)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(682..768)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     817..839
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     831..848
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     832..848
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    864..963
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1801..1900)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1917..1936)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1918..1936)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1941..1957)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1941..1957)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2059..2078)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2070..2879
                     /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(2137..2156)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3026..3381,1..233)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
       61 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
      121 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
      181 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
      241 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
      301 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
      361 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
      421 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
      481 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt accgctagcc
      541 aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttagtttg
      601 atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac
      661 gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
      721 gataaaacga aaggcccagt cttccgactg agcctttcgt tttatttgat gcctggcagt
      781 tccctactct cgcgttaacg ctagcatgga tgttttccca gtcacgacgt tgtaaaacga
      841 cggccagtct taagctcggg ccccaaataa tgattttatt ttgactgata gtgacctgtt
      901 cgttgcaaca aattgatgag caatgctttt ttataatgcc aactttgtac aaaaaagcag
      961 gcttcgaagg agatagaacc atggggcctg aaggaaatgt ggatgttgag ttgattgata
     1021 agagcacaaa cagatacagc gtttggttcc ccactgctgg ctggtatctg tggtcagcca
     1081 caggcctcgg cttcctggta agggatgagg tcacagttac catcgccttc ggcagctgga
     1141 gtcagcacct ggccctggac ctgcagcacc atgaacagtg gctggtgggc ggccccttgt
     1201 ttgatgtcac tgcagagcca gaggaggctg tcgccgaaat ccacctcccc cacttcatct
     1261 ccctccaagc aggtgaggtg gacgtctcct ggtttctcgt tgcccatttt aagaatgaag
     1321 ggatggtcct ggagcatcca gcccgggtgg agcctttcta tgctgtcctg gaaagcccca
     1381 gcttctctct gatgggcatc ctgctgcgga tcgccagtgg gactcgcctc tccatcccca
     1441 tcacttccaa cacattgatc tattatcacc cccaccccga agatattaag ttccacttgt
     1501 accttgtccc cagcgacgcc ttgctaacaa aggcgataga tgatgaggaa gatcgcttcc
     1561 atggtgtgcg cctgcagact tcgcccccaa tggaacccct gaactttggt tccagttata
     1621 ttgtgtctaa ttctgctaac ctgaaagtaa tgcccaagga gttgaaattg tcctacagga
     1681 gccctggaga aattcagcac ttctcaaaag gctatgctgg gcagatgaag gaacccattc
     1741 aacttgagat tactgaaaaa agacatggga ctttggtgtg ggatactgag gtgaagccag
     1801 acccagcttt cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca
     1861 acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct
     1921 atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa
     1981 atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg
     2041 cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc
     2101 cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg
     2161 tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt
     2221 ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa
     2281 actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg
     2341 atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat
     2401 atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt
     2461 cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc
     2521 aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct
     2581 ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag
     2641 tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag
     2701 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat
     2761 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta
     2821 ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat
     2881 cagaattggt taattggttg taacactggc agagcattac gctgacttga cgggacggcg
     2941 caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc
     3001 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3061 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3121 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     3181 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3241 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3301 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3361 cacagcccag cttggagcga a
//
