LOCUS       sequence-332713-        3381 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pDONR221 for Gateway shuttling. Contains the FIIND (L162M-P434, no 
            stop codon).
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
FEATURES             Location/Qualifiers
     source          1..3381
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(137..164)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(256..342)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     391..413
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     405..422
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     406..422
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    438..537
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1375..1474)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1491..1510)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1492..1510)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1515..1531)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1515..1531)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1633..1652)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1644..2453
                     /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(1711..1730)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2600..3188
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3089..3108
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3342..3359
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
       61 ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag
      121 agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct
      181 ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa
      241 atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa
      301 acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta
      361 ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa acgacggcca
      421 gtcttaagct cgggccccaa ataatgattt tattttgact gatagtgacc tgttcgttgc
      481 aacaaattga tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa gcaggcttcg
      541 aaggagatag aaccatgggg cctgaaggaa atgtggatgt tgagttgatt gataagagca
      601 caaacagata cagcgtttgg ttccccactg ctggctggta tctgtggtca gccacaggcc
      661 tcggcttcct ggtaagggat gaggtcacag tgacgattgc gtttggttcc tggagtcagc
      721 acctggccct ggacctgcag caccatgaac agtggctggt gggcggcccc ttgtttgatg
      781 tcactgcaga gccagaggag gctgtcgccg aaatccacct cccccacttc atctccctcc
      841 aagcaggtga ggtggacgtc tcctggtttc tcgttgccca ttttaagaat gaagggatgg
      901 tcctggagca tccagcccgg gtggagcctt tctatgctgt cctggaaagc cccagcttct
      961 ctctgatggg catcctgctg cggatcgcca gtgggactcg cctctccatc cccatcactt
     1021 ccaacacatt gatctattat cacccccacc ccgaagatat taagttccac ttgtaccttg
     1081 tccccagcga cgccttgcta acaaaggcga tagatgatga ggaagatcgc ttccatggtg
     1141 tgcgcctgca gacttcgccc ccaatggaac ccctgaactt tggttccagt tatattgtgt
     1201 ctaattctgc taacctgaaa gtaatgccca aggagttgaa attgtcctac aggagccctg
     1261 gagaaattca gcacttctca aaattctatg ctgggcagat gaaggaaccc attcaacttg
     1321 agattactga aaaaagacat gggactttgg tgtgggatac tgaggtgaag ccagacccag
     1381 ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac
     1441 aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg
     1501 agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct
     1561 gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca
     1621 taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat
     1681 taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc
     1741 aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga
     1801 aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc
     1861 tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat
     1921 ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg
     1981 attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc
     2041 ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac
     2101 gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg
     2161 ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca
     2221 ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta
     2281 ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact
     2341 gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata
     2401 atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat
     2461 tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct
     2521 catgaccaaa atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag
     2581 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     2641 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     2701 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     2761 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     2821 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     2881 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     2941 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     3001 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     3061 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     3121 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3181 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     3241 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     3301 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     3361 aagcggaaga gcgcccaata c
//
