LOCUS       sequence-432144-        5664 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway shuttling vector for cloning N-terminal NLRP3 tags (contains
            a stop codon--note: bulky N-terminal tags are likely not 
            functional).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5664)
  AUTHORS   Hollingsworth LR, Veeraraghavan P, Paulo JA, Harper JW
  TITLE     Spatiotemporal proteomic profiling of cellular responses to NLRP3 
            agonists
  JOURNAL   bioRxiv 2024.04.19.590338
REFERENCE   2  (bases 1 to 5664)
  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"; journalName: "bioRxiv 
            2024.04.19.590338"
COMMENT     Sequence Label: pDONR221 msNLRP3 (STOP)
FEATURES             Location/Qualifiers
     source          1..5664
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..100
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3221..3320)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3337..3356)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3338..3356)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3361..3377)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3361..3377)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3479..3498)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3490..4299
                     /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(3557..3576)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4446..5034
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4935..4954
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5188..5205
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5364..5391)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5483..5569)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5618..5640
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5632..5649
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5633..5649
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
       61 tgctttttta taatgccaac tttgtacaaa aaagcaggct tcgaaggaga tagaaccatg
      121 acgagtgtcc gttgcaagct ggctcagtat ctagaggacc ttgaagatgt ggacctcaag
      181 aaattcaaaa tgcatttgga agattacccg cccgagaaag gctgtatccc agtccccagg
      241 ggccagatgg agaaggcaga tcacttggat ctagccacac tcatgattga cttcaatggc
      301 gaggagaagg cctgggccat ggctgtgtgg atctttgctg cgatcaacag gcgagacctc
      361 tgggaaaaag ctaagaagga ccagccagag tggaatgaca cgtgtacatc acattcctct
      421 atggtatgcc aggaggacag ccttgaagaa gagtggatgg gtttgctggg atatctctcc
      481 cgcatctcca tttgtaaaaa gaagaaagat tactgtaaga tgtacagacg acatgtgaga
      541 agcaggttct actctatcaa ggacaggaac gcgcgtctag gtgagagtgt ggacctcaac
      601 agtcgctaca cgcagctcca actggtcaag gagcatccaa gcaagcagga gcgggagcat
      661 gaactcctga ccatcggccg gactaaaatg cgggacagcc ccatgagttc ccttaagctg
      721 gagctgctgt ttgagcccga ggacgggcac tcggagcctg tgcacacagt ggtgttccag
      781 ggagcagcag gcatcgggaa aaccatccta gccaggaaga ttatgttgga ctgggcactg
      841 ggaaagctct tcaaagacaa atttgactat ttgttcttta tccactgccg agaggtgagc
      901 ctcaggacgc caaggagtct agcagacctg attgtcagct gctggcctga cccaaaccca
      961 ccagtgtgca agatcctgcg caagccttcc aggatcctct tcctcatgga tggctttgat
     1021 gagctacaag gggcctttga cgagcacatt ggggaggtct gcacagactg gcaaaaggct
     1081 gtgcggggag acattctgct aagcagcctc atccgaaaga aactgctgcc caaggcctct
     1141 ctgctcataa cgacgaggcc ggtagccttg gagaaactgc agcatctcct ggaccacccc
     1201 cgccatgtgg agatcctagg tttctctgag gccaaaagga aggagtattt ctttaagtat
     1261 ttctccaacg agctgcaggc ccgggaggcc ttcaggctga tccaagagaa tgaggtcctc
     1321 tttaccatgt gcttcatccc cctggtctgc tggattgtgt gcacggggct aaagcaacag
     1381 atggagaccg ggaagagcct ggcccagacc tccaagacca ctacggccgt ctacgtcttc
     1441 ttcctttcca gcctgctgca atcccggggg ggcattgagg agcatctctt ctctgactac
     1501 ctacaggggc tctgttcact ggctgcggat ggaatttgga accagaaaat cctatttgag
     1561 gagtgtgatc tgcggaagca cggcctgcag aagactgacg tctccgcttt cctgaggatg
     1621 aacgtgttcc agaaggaagt ggactgcgag agattctaca gcttcagcca catgactttc
     1681 caggagttct tcgctgctat gtactatttg ctggaagagg aggcagaggg ggagaccgtg
     1741 aggaaaggac caggaggttg ttcagatctt ctgaaccgag acgtgaaggt cctactagaa
     1801 aattacggca agtttgaaaa aggctatctg atttttgttg tccgattcct ctttggcctt
     1861 gtaaaccagg agagaacctc ttatttggag aagaaactaa gttgcaagat ctctcagcaa
     1921 gtcagactgg aactactgaa gtggattgaa gtgaaagcca aggccaagaa gctgcagtgg
     1981 cagcccagcc aactggaact gttctactgc ctgtacgaga tgcaggagga agactttgtg
     2041 cagagtgcca tggaccactt tcccaaaatt gagatcaacc tctctaccag aatggaccac
     2101 gtggtttcct ccttttgtat taagaactgt catagggtca aaacgctttc cctgggtttt
     2161 tttcacaact cgcccaagga ggaagaagaa gagaggagag gaggtcgacc cttggaccag
     2221 gttcagtgtg ttttcccaga cactcatgtt gcctgttctt ccagactggt gaactgctgc
     2281 ctcacttcta gcttctgccg tggtctcttc tcaagtctaa gcaccaaccg gagcctcact
     2341 gaactggacc tcagtgacaa tactctggga gacccgggca tgagggtgct gtgtgaggca
     2401 ctccagcacc caggctgtaa cattcagaga ctgtggttgg ggcgctgcgg actgtcccat
     2461 caatgctgct tcgacatctc ctctgtcctg agcagcagcc agaagctggt ggagctggac
     2521 ctcagtgaca atgccctggg ggactttgga atcagattgc tgtgtgtggg actgaagcac
     2581 ctgctctgca acctccagaa actgtggttg gtgagctgct gtctcacatc cgcgtgttgt
     2641 caggatctcg cattggttct gagctccaac cattctctga ccagactgta cattggagaa
     2701 aatgccttgg gagactcagg agtccaagtt ttgtgtgaaa agatgaagga cccacagtgt
     2761 aacttgcaga agctggggtt ggtgaattcc ggccttactt caatctgttg ttcagctctg
     2821 acctctgtgc tcaaaaccaa ccagaacttc acacacctct atctacgaag caatgccctt
     2881 ggagacacag gactcaggct cctctgtgag gggcttctgc acccggactg taaactacag
     2941 atgctggaat tagacaactg cagcctcacc tcacacagct gctggaatct ctccacaatt
     3001 ctgacccaca accacagcct tcggaagctg aacctgggca acaatgatct tggcgatctg
     3061 tgcgtggtga ccctctgtga ggtgctgaaa cagcagggct gcctcctgca gagcctacag
     3121 ttgggtgaaa tgtacttaaa tcgtgaaaca aaacgtgcct tagaagcgct ccaggaagaa
     3181 aagcctgagc tgactatagt cttcgagatt tcctggtagg acccagcttt cttgtacaaa
     3241 gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt
     3301 caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc gtattacatg
     3361 gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca
     3421 caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca
     3481 aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg
     3541 gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca
     3601 atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt
     3661 agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg
     3721 cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact
     3781 gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat
     3841 attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt
     3901 ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt
     3961 ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg
     4021 aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc
     4081 tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga
     4141 gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt
     4201 tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat
     4261 aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg
     4321 taacactggc agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc
     4381 cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4441 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4501 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4561 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     4621 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4681 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     4741 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     4801 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     4861 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     4921 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     4981 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     5041 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     5101 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     5161 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     5221 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     5281 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc
     5341 caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttagttt
     5401 gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa
     5461 cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac
     5521 agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga tgcctggcag
     5581 ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg ttgtaaaacg
     5641 acggccagtc ttaagctcgg gccc
//
