LOCUS       sequence-441242-        5661 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway shuttling vector for cloning C-terminal NLRP3 tags (contains
            no stop codon)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5661)
  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 5661)
  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
FEATURES             Location/Qualifiers
     source          1..5661
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3786..3885)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3902..3921)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3903..3921)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3926..3942)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3926..3942)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4044..4063)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4055..4864
                     /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(4122..4141)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5011..5599
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5500..5519
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggcttc gaaggagata gaaccatgac gagtgtccgt tgcaagctgg ctcagtatct
      721 agaggacctt gaagatgtgg acctcaagaa attcaaaatg catttggaag attacccgcc
      781 cgagaaaggc tgtatcccag tccccagggg ccagatggag aaggcagatc acttggatct
      841 agccacactc atgattgact tcaatggcga ggagaaggcc tgggccatgg ctgtgtggat
      901 ctttgctgcg atcaacaggc gagacctctg ggaaaaagct aagaaggacc agccagagtg
      961 gaatgacacg tgtacatcac attcctctat ggtatgccag gaggacagcc ttgaagaaga
     1021 gtggatgggt ttgctgggat atctctcccg catctccatt tgtaaaaaga agaaagatta
     1081 ctgtaagatg tacagacgac atgtgagaag caggttctac tctatcaagg acaggaacgc
     1141 gcgtctaggt gagagtgtgg acctcaacag tcgctacacg cagctccaac tggtcaagga
     1201 gcatccaagc aagcaggagc gggagcatga actcctgacc atcggccgga ctaaaatgcg
     1261 ggacagcccc atgagttccc ttaagctgga gctgctgttt gagcccgagg acgggcactc
     1321 ggagcctgtg cacacagtgg tgttccaggg agcagcaggc atcgggaaaa ccatcctagc
     1381 caggaagatt atgttggact gggcactggg aaagctcttc aaagacaaat ttgactattt
     1441 gttctttatc cactgccgag aggtgagcct caggacgcca aggagtctag cagacctgat
     1501 tgtcagctgc tggcctgacc caaacccacc agtgtgcaag atcctgcgca agccttccag
     1561 gatcctcttc ctcatggatg gctttgatga gctacaaggg gcctttgacg agcacattgg
     1621 ggaggtctgc acagactggc aaaaggctgt gcggggagac attctgctaa gcagcctcat
     1681 ccgaaagaaa ctgctgccca aggcctctct gctcataacg acgaggccgg tagccttgga
     1741 gaaactgcag catctcctgg accacccccg ccatgtggag atcctaggtt tctctgaggc
     1801 caaaaggaag gagtatttct ttaagtattt ctccaacgag ctgcaggccc gggaggcctt
     1861 caggctgatc caagagaatg aggtcctctt taccatgtgc ttcatccccc tggtctgctg
     1921 gattgtgtgc acggggctaa agcaacagat ggagaccggg aagagcctgg cccagacctc
     1981 caagaccact acggccgtct acgtcttctt cctttccagc ctgctgcaat cccggggggg
     2041 cattgaggag catctcttct ctgactacct acaggggctc tgttcactgg ctgcggatgg
     2101 aatttggaac cagaaaatcc tatttgagga gtgtgatctg cggaagcacg gcctgcagaa
     2161 gactgacgtc tccgctttcc tgaggatgaa cgtgttccag aaggaagtgg actgcgagag
     2221 attctacagc ttcagccaca tgactttcca ggagttcttc gctgctatgt actatttgct
     2281 ggaagaggag gcagaggggg agaccgtgag gaaaggacca ggaggttgtt cagatcttct
     2341 gaaccgagac gtgaaggtcc tactagaaaa ttacggcaag tttgaaaaag gctatctgat
     2401 ttttgttgtc cgattcctct ttggccttgt aaaccaggag agaacctctt atttggagaa
     2461 gaaactaagt tgcaagatct ctcagcaagt cagactggaa ctactgaagt ggattgaagt
     2521 gaaagccaag gccaagaagc tgcagtggca gcccagccaa ctggaactgt tctactgcct
     2581 gtacgagatg caggaggaag actttgtgca gagtgccatg gaccactttc ccaaaattga
     2641 gatcaacctc tctaccagaa tggaccacgt ggtttcctcc ttttgtatta agaactgtca
     2701 tagggtcaaa acgctttccc tgggtttttt tcacaactcg cccaaggagg aagaagaaga
     2761 gaggagagga ggtcgaccct tggaccaggt tcagtgtgtt ttcccagaca ctcatgttgc
     2821 ctgttcttcc agactggtga actgctgcct cacttctagc ttctgccgtg gtctcttctc
     2881 aagtctaagc accaaccgga gcctcactga actggacctc agtgacaata ctctgggaga
     2941 cccgggcatg agggtgctgt gtgaggcact ccagcaccca ggctgtaaca ttcagagact
     3001 gtggttgggg cgctgcggac tgtcccatca atgctgcttc gacatctcct ctgtcctgag
     3061 cagcagccag aagctggtgg agctggacct cagtgacaat gccctggggg actttggaat
     3121 cagattgctg tgtgtgggac tgaagcacct gctctgcaac ctccagaaac tgtggttggt
     3181 gagctgctgt ctcacatccg cgtgttgtca ggatctcgca ttggttctga gctccaacca
     3241 ttctctgacc agactgtaca ttggagaaaa tgccttggga gactcaggag tccaagtttt
     3301 gtgtgaaaag atgaaggacc cacagtgtaa cttgcagaag ctggggttgg tgaattccgg
     3361 ccttacttca atctgttgtt cagctctgac ctctgtgctc aaaaccaacc agaacttcac
     3421 acacctctat ctacgaagca atgcccttgg agacacagga ctcaggctcc tctgtgaggg
     3481 gcttctgcac ccggactgta aactacagat gctggaatta gacaactgca gcctcacctc
     3541 acacagctgc tggaatctct ccacaattct gacccacaac cacagccttc ggaagctgaa
     3601 cctgggcaac aatgatcttg gcgatctgtg cgtggtgacc ctctgtgagg tgctgaaaca
     3661 gcagggctgc ctcctgcaga gcctacagtt gggtgaaatg tacttaaatc gtgaaacaaa
     3721 acgtgcctta gaagcgctcc aggaagaaaa gcctgagctg actatagtct tcgagatttc
     3781 ctgggaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     3841 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     3901 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     3961 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     4021 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     4081 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     4141 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     4201 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     4261 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     4321 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     4381 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     4441 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     4501 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     4561 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     4621 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     4681 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     4741 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     4801 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     4861 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     4921 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     4981 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5041 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5101 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5161 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5221 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5281 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5341 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5401 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5461 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5521 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5581 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5641 ctggcctttt gctcacatgt t
//
