LOCUS       sequence-432153-        5289 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway shuttling vector for cloning N- and C-terminal 
            NLRP3(Delta-PYD, I125M-END) tags (contains no stop codon).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5289)
  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 5289)
  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(Delta-PYD, I125M-END)
FEATURES             Location/Qualifiers
     source          1..5289
                     /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(2846..2945)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2962..2981)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2963..2981)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2986..3002)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2986..3002)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3104..3123)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3115..3924
                     /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(3182..3201)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4071..4659
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4560..4579
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4813..4830
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4989..5016)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5108..5194)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5243..5265
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5257..5274
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5258..5274
                     /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 tgtaaaaaga agaaagatta ctgtaagatg tacagacgac atgtgagaag caggttctac
      181 tctatcaagg acaggaacgc gcgtctaggt gagagtgtgg acctcaacag tcgctacacg
      241 cagctccaac tggtcaagga gcatccaagc aagcaggagc gggagcatga actcctgacc
      301 atcggccgga ctaaaatgcg ggacagcccc atgagttccc ttaagctgga gctgctgttt
      361 gagcccgagg acgggcactc ggagcctgtg cacacagtgg tgttccaggg agcagcaggc
      421 atcgggaaaa ccatcctagc caggaagatt atgttggact gggcactggg aaagctcttc
      481 aaagacaaat ttgactattt gttctttatc cactgccgag aggtgagcct caggacgcca
      541 aggagtctag cagacctgat tgtcagctgc tggcctgacc caaacccacc agtgtgcaag
      601 atcctgcgca agccttccag gatcctcttc ctcatggatg gctttgatga gctacaaggg
      661 gcctttgacg agcacattgg ggaggtctgc acagactggc aaaaggctgt gcggggagac
      721 attctgctaa gcagcctcat ccgaaagaaa ctgctgccca aggcctctct gctcataacg
      781 acgaggccgg tagccttgga gaaactgcag catctcctgg accacccccg ccatgtggag
      841 atcctaggtt tctctgaggc caaaaggaag gagtatttct ttaagtattt ctccaacgag
      901 ctgcaggccc gggaggcctt caggctgatc caagagaatg aggtcctctt taccatgtgc
      961 ttcatccccc tggtctgctg gattgtgtgc acggggctaa agcaacagat ggagaccggg
     1021 aagagcctgg cccagacctc caagaccact acggccgtct acgtcttctt cctttccagc
     1081 ctgctgcaat cccggggggg cattgaggag catctcttct ctgactacct acaggggctc
     1141 tgttcactgg ctgcggatgg aatttggaac cagaaaatcc tatttgagga gtgtgatctg
     1201 cggaagcacg gcctgcagaa gactgacgtc tccgctttcc tgaggatgaa cgtgttccag
     1261 aaggaagtgg actgcgagag attctacagc ttcagccaca tgactttcca ggagttcttc
     1321 gctgctatgt actatttgct ggaagaggag gcagaggggg agaccgtgag gaaaggacca
     1381 ggaggttgtt cagatcttct gaaccgagac gtgaaggtcc tactagaaaa ttacggcaag
     1441 tttgaaaaag gctatctgat ttttgttgtc cgattcctct ttggccttgt aaaccaggag
     1501 agaacctctt atttggagaa gaaactaagt tgcaagatct ctcagcaagt cagactggaa
     1561 ctactgaagt ggattgaagt gaaagccaag gccaagaagc tgcagtggca gcccagccaa
     1621 ctggaactgt tctactgcct gtacgagatg caggaggaag actttgtgca gagtgccatg
     1681 gaccactttc ccaaaattga gatcaacctc tctaccagaa tggaccacgt ggtttcctcc
     1741 ttttgtatta agaactgtca tagggtcaaa acgctttccc tgggtttttt tcacaactcg
     1801 cccaaggagg aagaagaaga gaggagagga ggtcgaccct tggaccaggt tcagtgtgtt
     1861 ttcccagaca ctcatgttgc ctgttcttcc agactggtga actgctgcct cacttctagc
     1921 ttctgccgtg gtctcttctc aagtctaagc accaaccgga gcctcactga actggacctc
     1981 agtgacaata ctctgggaga cccgggcatg agggtgctgt gtgaggcact ccagcaccca
     2041 ggctgtaaca ttcagagact gtggttgggg cgctgcggac tgtcccatca atgctgcttc
     2101 gacatctcct ctgtcctgag cagcagccag aagctggtgg agctggacct cagtgacaat
     2161 gccctggggg actttggaat cagattgctg tgtgtgggac tgaagcacct gctctgcaac
     2221 ctccagaaac tgtggttggt gagctgctgt ctcacatccg cgtgttgtca ggatctcgca
     2281 ttggttctga gctccaacca ttctctgacc agactgtaca ttggagaaaa tgccttggga
     2341 gactcaggag tccaagtttt gtgtgaaaag atgaaggacc cacagtgtaa cttgcagaag
     2401 ctggggttgg tgaattccgg ccttacttca atctgttgtt cagctctgac ctctgtgctc
     2461 aaaaccaacc agaacttcac acacctctat ctacgaagca atgcccttgg agacacagga
     2521 ctcaggctcc tctgtgaggg gcttctgcac ccggactgta aactacagat gctggaatta
     2581 gacaactgca gcctcacctc acacagctgc tggaatctct ccacaattct gacccacaac
     2641 cacagccttc ggaagctgaa cctgggcaac aatgatcttg gcgatctgtg cgtggtgacc
     2701 ctctgtgagg tgctgaaaca gcagggctgc ctcctgcaga gcctacagtt gggtgaaatg
     2761 tacttaaatc gtgaaacaaa acgtgcctta gaagcgctcc aggaagaaaa gcctgagctg
     2821 actatagtct tcgagatttc ctgggaccca gctttcttgt acaaagttgg cattataaga
     2881 aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt
     2941 atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc
     3001 tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata
     3061 tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc
     3121 catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat
     3181 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat
     3241 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     3301 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     3361 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa
     3421 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     3481 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
     3541 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     3601 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
     3661 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     3721 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     3781 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     3841 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     3901 ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc
     3961 attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac
     4021 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4081 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4141 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4201 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     4261 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4321 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4381 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     4441 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     4501 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4561 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     4621 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     4681 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     4741 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     4801 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     4861 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     4921 aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta
     4981 gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt
     5041 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc
     5101 ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
     5161 cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgcg
     5221 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaag
     5281 ctcgggccc
//
