LOCUS       sequence-450382-        3860 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DONR vector for inflammasome speck reporter, designed from 
            PMC9623368..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3860)
  TITLE     Harper lab Rab11 and inflammasome reporter plasmids
REFERENCE   2  (bases 1 to 3860)
  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"
COMMENT     Sequence Label: pDONR223 CASP1[CARD]-mSG2 (inflammasome reporter)
FEATURES             Location/Qualifiers
     source          1..3860
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(919..946)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1038..1124)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1173..1195
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1187..1204
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1188..1204
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1220..1319
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      1322..1331
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1685..2362
                     /codon_start=1
                     /product="bright and highly photostable monomeric green 
                     fluorescent protein (Ando et al., 2023)"
                     /label=mStayGold(J)
                     /note="includes multiple mutations plus an N-terminal 
                     iinker sequence"
                     /translation="MVSTGEELFTGVVPFKFQLKGTINGKSFTVEGEGEGNSHEGSHKG
                     KYVCTSGKLPMSWAALGTSFGYGMKYYTKYPSGLKNWFHEVMPEGFTYDRHIQYKGDGS
                     IHAKHQHFMKNGTYHNIVEFTGQDFKENSPVLTGDMDVSLPNEVQHIPRDDGVECTVTL
                     TYPLLSDESKCVEAYQNTIIKPLHNQPAPDVPYHWIRKQYTQSKDDTEERDHIIQSETL
                     EAHL"
     protein_bind    complement(2390..2489)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2506..2525)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2507..2525)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2530..2546)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2530..2546)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2977..3768
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga
      901 agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc
      961 ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc
     1021 aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata
     1081 aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc
     1141 tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc
     1201 cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga cctgttcgtt
     1261 gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa aagcaggctc
     1321 cgccaccatg gccgacaagg tcctgaagga gaagagaaag ctgtttatcc gttccatggg
     1381 tgaaggtaca ataaatggct tactggatga attattacag acaagggtgc tgaacaagga
     1441 agagatggag aaagtaaaac gtgaaaatgc tacagttatg gataagaccc gagctttgat
     1501 tgactccgtt attccgaaag gggcacaggc atgccaaatt tgcatcacat acatttgtga
     1561 agaagacagt tacctggcag ggacgctggg actctcagca gatcaaacat ctggaaatta
     1621 ccttaatatg caagactctc aaggagtact ttcttccttt ccagctcctc aggcagtgca
     1681 ggacatggtg tctacaggcg aggagctgtt taccggcgtg gtgcccttca agttccagct
     1741 gaagggcacc atcaacggca agagcttcac cgtggaaggc gagggcgagg gcaatagcca
     1801 cgagggcagc cacaaaggca agtacgtgtg caccagcggc aaactgccaa tgtcttgggc
     1861 cgccctggga actagcttcg gctatggcat gaagtactac accaagtacc ccagcggcct
     1921 gaagaactgg ttccacgagg tgatgcccga gggcttcacc tacgacagac acatccagta
     1981 caagggcgac ggcagcatcc acgccaagca ccagcacttc atgaagaacg gcacctacca
     2041 caacatcgtg gagttcaccg gccaggactt caaggagaac agccccgtgc tgaccggcga
     2101 catggacgtg agcctgccca acgaggtgca gcacatcccc agagatgacg gcgtggagtg
     2161 cacagtgacc ctgacgtacc ctctgctgag cgacgagagc aagtgcgtgg aggcctacca
     2221 gaacaccatc atcaagcccc tgcacaatca gccagccccc gatgtgccat accactggat
     2281 cagaaagcag tacacccaga gcaaggacga caccgaggag agagaccaca tcatccagag
     2341 cgagaccctg gaggcccacc tgtactccag gacgaagctg gagtaatgaa cccagctttc
     2401 ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc
     2461 actatcagtc aaaataaaat cattatttgc catccagctg atatccccta tagtgagtcg
     2521 tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt
     2581 tacattgcac aagataaaaa tatatcatca tgcctcctct agaccagcca ggacagaaat
     2641 gcctcgactt cgctgctacc caaggttgcc gggtgacgca caccgtggaa acggatgaag
     2701 gcacgaaccc agtggacata agcctgttcg gttcgtaagc tgtaatgcaa gtagcgtatg
     2761 cgctcacgca actggtccag aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg
     2821 gttttcatgg cttgttatga ctgttttttt ggggtacagt ctatgcctcg ggcatccaag
     2881 cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg
     2941 cagcagggca gtcgccctaa aacaaagtta aacattatga gggaagcggt gatcgccgaa
     3001 gtatcgactc aactatcaga ggtagttggc gtcatcgagc gccatctcga accgacgttg
     3061 ctggccgtac atttgtacgg ctccgcagtg gatggcggcc tgaagccaca cagtgatatt
     3121 gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac
     3181 gaccttttgg aaacttcggc ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc
     3241 accattgttg tgcacgacga catcattccg tggcgttatc cagctaagcg cgaactgcaa
     3301 tttggagaat ggcagcgcaa tgacattctt gcaggtatct tcgagccagc cacgatcgac
     3361 attgatctgg ctatcttgct gacaaaagca agagaacata gcgttgcctt ggtaggtcca
     3421 gcggcggagg aactctttga tccggttcct gaacaggatc tatttgaggc gctaaatgaa
     3481 accttaacgc tatggaactc gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt
     3541 acgttgtccc gcatttggta cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct
     3601 gccgactggg caatggagcg cctgccggcc cagtatcagc ccgtcatact tgaagctaga
     3661 caggcttatc ttggacaaga agaagatcgc ttggcctcgc gcgcagatca gttggaagaa
     3721 tttgtccact acgtgaaagg cgagatcacc aaggtagtcg gcaaataacc ctcgagccac
     3781 ccatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta
     3841 gaaaagatca aaggatcttc
//
