LOCUS       sequence-343152-        3690 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pDONR221 for Gateway shuttling. Contains the CARD8 FIIND and CARD 
            (L162M-L537, no stop codon).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3690)
  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 3690)
  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 ZUC
FEATURES             Location/Qualifiers
     source          1..3690
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     101..120
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     354..371
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(530..557)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(649..735)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     784..806
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     798..815
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     799..815
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    831..930
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2077..2176)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2193..2212)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2194..2212)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2217..2233)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2217..2233)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2335..2354)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2346..3155
                     /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(2413..2432)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(3302..3690,1..200)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
       61 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
      121 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
      181 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
      241 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
      301 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
      361 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
      421 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
      481 acgcaattaa tacgcgtacc gctagccagg aagagtttgt agaaacgcaa aaaggccatc
      541 cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc gtcctgcccg
      601 ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat ttgtcctact
      661 caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt ccgactgagc
      721 ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta gcatggatgt
      781 tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc caaataatga
      841 ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa tgctttttta
      901 taatgccaac tttgtacaaa aaagcaggct tcgaaggaga tagaaccatg gggcctgaag
      961 gaaatgtgga tgttgagttg attgataaga gcacaaacag atacagcgtt tggttcccca
     1021 ctgctggctg gtatctgtgg tcagccacag gcctcggctt cctggtaagg gatgaggtca
     1081 cagtgacgat tgcgtttggt tcctggagtc agcacctggc cctggacctg cagcaccatg
     1141 aacagtggct ggtgggcggc cccttgtttg atgtcactgc agagccagag gaggctgtcg
     1201 ccgaaatcca cctcccccac ttcatctccc tccaagcagg tgaggtggac gtctcctggt
     1261 ttctcgttgc ccattttaag aatgaaggga tggtcctgga gcatccagcc cgggtggagc
     1321 ctttctatgc tgtcctggaa agccccagct tctctctgat gggcatcctg ctgcggatcg
     1381 ccagtgggac tcgcctctcc atccccatca cttccaacac attgatctat tatcaccccc
     1441 accccgaaga tattaagttc cacttgtacc ttgtccccag cgacgccttg ctaacaaagg
     1501 cgatagatga tgaggaagat cgcttccatg gtgtgcgcct gcagacttcg cccccaatgg
     1561 aacccctgaa ctttggttcc agttatattg tgtctaattc tgctaacctg aaagtaatgc
     1621 ccaaggagtt gaaattgtcc tacaggagcc ctggagaaat tcagcacttc tcaaaattct
     1681 atgctgggca gatgaaggaa cccattcaac ttgagattac tgaaaaaaga catgggactt
     1741 tggtgtggga tactgaggtg aagccagtgg atctccagct tgtagctgca tcagcccctc
     1801 ctcctttctc aggtgcagcc tttgtgaagg agaaccaccg gcaactccaa gccaggatgg
     1861 gggacctgaa aggggtgctc gatgatctcc aagacaatga ggttcttact gagaatgaga
     1921 aggagctggt ggagcaggaa aagacacggc agagcaagaa tgaggccttg ctgagcatgg
     1981 tggagaagaa aggggacctg gccctggacg tgctcttcag aagcattagt gaaagggacc
     2041 cttacctcgt gtcctatctt agacagcaga atttggaccc agctttcttg tacaaagttg
     2101 gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa
     2161 ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca
     2221 tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag
     2281 ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg
     2341 gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg
     2401 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
     2461 atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg
     2521 ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc
     2581 ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga
     2641 tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg
     2701 ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt
     2761 ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg
     2821 ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
     2881 aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
     2941 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
     3001 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
     3061 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
     3121 tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac
     3181 actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta
     3241 acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3301 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3361 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3421 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     3481 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3541 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3601 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3661 cctacaccga actgagatac ctacagcgtg
//
