LOCUS       sequence-430372-        6601 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector with mouse EEA1 .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6601)
  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 6601)
  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: pTwist-msEEA1
FEATURES             Location/Qualifiers
     source          1..6601
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(2332..2431)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2488..2504)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2488..2504)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2501..2523)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(2676..3264)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2756..2775)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3400..4215)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3400..4215)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4123..4142
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4207..4226
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     4336..4352
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4336..4352
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      4353..4399
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4513..4535
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4527..4544
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4528..4544
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4597..4696
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 aaggtctcac aactcgacat ccaaatcaag gccaagactg agctccttct atccgcagag
       61 gccgctaagg ccgcccaaag ggccgacctg caaaatcacc tagacactgc acaacacgct
      121 ctccaagaca agcaacaaga gctcaataag gtctccgtcc aactagacca actcacagca
      181 aagttccaag agaagcaaga gcactgtatc caacttgagt cccacctcaa ggaccacaag
      241 gagaagcacc tctccctcga gcaaaaggtc gaggaccttg agggtcacat aaagaagctc
      301 gaggccgacg ccctagaggt taaggcctca aaggagcaag cgctccaatc cctgcaacaa
      361 caacggcaac tcagtactga cctagagctt agaaatgcag agctaagtcg ggagcttcaa
      421 gagcaagaag aggttgtctc ttgtactaag cttgaccttc aaaataagtc tgagatcctc
      481 gagaatataa agcaaacatt gacaaagaag gaagaggaga atgtcgttct taagcaagag
      541 ttcgagaagc tatcccaaga ctccaagact caacacaagg agttgggaga ccggatgcaa
      601 gccgcagtaa ctgagctcac agctgtaaag gcacaaaagg acgcactcct tgccgagctc
      661 agtactacga aggagaagct atccaaggtt tcagactctc tcaagaattc taagtccgag
      721 ttcgagaagg agaatcaaaa gggaaaggct gctgttcttg acctagagaa ggcatgtaag
      781 gaactcaagc accaactcca agtccaagcc gagtccgcat tgaaggagca agaggacttg
      841 aagaagtccc tcgagaagga gaaggagact tcccaacaat tgaagatcga gctcaattca
      901 gttaagggtg aggtcagtca agcccaaaat acattgaagc aaaaggagaa ggacgagcaa
      961 caacttcaag gaacaataaa tcaactcaag caaagtgccg agcaaaagaa gaagcaaatc
     1021 gaggcactac aaggcgaggt taagaatgca gtttctcaaa agactgtcct cgagaataag
     1081 ttgcaacaac aatctagtca agcagcacaa gagttggctg ccgagaaggg taagctaagt
     1141 gcccttcaat ctaattacga gaagtgtcaa gccgacctca agcaactcca atctgacctt
     1201 tacggtaagg aatcagagtt gctagcaact aggcaagacc taaagtccgt cgaggagaag
     1261 ttgacattgg cacaagagga cctaatctct aatagaaatc aaataggaaa tcaaaataag
     1321 tctatccaag agttgcaagc tgccaaggct tctctcgagc aagactccgc aaagaaagag
     1381 gcccttctaa aggagcaatc caaggcactc gaggacgcac aaagagagaa gagtgtcaag
     1441 gagaaggagt tggttgctga gaagtccaag ttggcagaga tggaagagat caagtgtcgt
     1501 caagagaagg agatcactaa gctcaatgag gagttgaagt ctcacaagca agagtccata
     1561 aaggagatca cgaatctaaa ggacgccaag caactactaa tccaacaaaa gcttgagttg
     1621 caagggcggg tcgactctct caaggcagct ttggagcaag agaaggagtc tcaacaactc
     1681 atgagggagc aagtaaagaa ggaagaggag aagagaaaag aggagttcag tgagaaagag
     1741 gccaagcttc actctgagat caaggagaaa gaggctggta tgaagaagca cgaggagaat
     1801 gaggcaaagt tgacaatgca agtaacaact ctcaatgaga atttggggac ggtcaagaag
     1861 gagtggcaat cctctcaaag aagagtatcc gagcttgaga agcaaactga cgaccttcgg
     1921 ggagagatcg cagtacttga ggccactgtc caaaataatc aagacgagcg tagggctctc
     1981 ttggagcgat gtctcaaggg tgagggagag atcgagaagc tccaaactaa ggccctagag
     2041 ctacaacgta agctcgacaa tacaacggct gctgttcaag agttgggtcg tgagaatcaa
     2101 agtctccaaa tcaagcacac acaagctctt aatcgaaagt gggccgagga caatgaggtt
     2161 caaaattgta tgtcttgtgg taagtgtttc tccgtcacag ttcggagaca ccactgtagg
     2221 caatgtggta atatcttctg tgctgagtgt tccactaaga atgcacttac tccatcctca
     2281 aagaagcctg tccgagtctg tgacgcttgt ttcaatgacc tgcaaggtta gacccagctt
     2341 tcttgtacaa agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg
     2401 tcactatcag tcaaaataaa atcattattt gaggctaggt ggaggctcag tgatgataag
     2461 tctgcgatgg tggatgcatg tgtcatggtc atagctgttt cctgtgtgaa attgttatcc
     2521 gctcagaggg cacaatccta ttccgcgcta tccgacaatc tccaagacat taggtggagt
     2581 tcagttcggc gtatggcata tgtcgctgga aagaacatgt gagcaaaagg ccagcaaaag
     2641 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2701 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2761 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2821 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2881 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2941 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3001 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3061 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     3121 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3181 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3241 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3301 ctattcaaca aagccgccgt cccgtcaagt cagcgtaaat gggtaggggg cttcaaatcg
     3361 tccgctctgc cagtgttaca accaattaac aaattctgat tagaaaaact catcgagcat
     3421 caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg
     3481 tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta
     3541 tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa
     3601 aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa
     3661 aagcttatgc atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa
     3721 atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac
     3781 gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac
     3841 tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc
     3901 tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg
     3961 cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt
     4021 aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt
     4081 cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata
     4141 cccatataaa tcagcatcca tgttggaatt taatcgcggc ctcgagcaag acgtttcccg
     4201 ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt
     4261 tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag acacaacgtg
     4321 gctttccccc gccgctctag aactagtgga tccaaataaa acgaaaggct cagtcgaaag
     4381 actgggcctt tcgttttatc tgttgtttgt cgcattatac gagacgtcca ggttgggata
     4441 cctgaaacaa aacccatcgt acggccaagg aagtctccaa taactgtgat ccaccacaag
     4501 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtcatgca taatccgcac
     4561 gcatctggaa taaggaagtg ccattccgcc tgacctcaaa taatgatttt attttgactg
     4621 atagtgacct gttcgttgca acagattgat gagcaatgct tttttataat gccaactttg
     4681 tacaaaaaag caggctatgt tcagacgaat tctccaaaga actcctggtc gggtcgggtc
     4741 tcaaggtagt gacttggact catcagccac accaataaat acagtagacg taaataatga
     4801 gagttcatcc gaggggttca tctgtcctca atgtatgaag tccttgggat cagccgacga
     4861 gttgttcaag cactaccaag ctgtccacga cgccgggaat gactccggac acggtggcga
     4921 ggctggactc gccctcacga gggacgacat aactctgctc cggcaagagg tccaagacct
     4981 tcaagcctct ttgaaagagg agaagtggta ctcagaggag ttgaagaagg agcttgagaa
     5041 gtaccaagga ttgcaacaac aagaggccaa gtccgacgga ttggtaacag actcctctgc
     5101 agagttgcaa gcacttgagc aacaactaga agaggcccaa actgagaatt tcaatataaa
     5161 gcaaatgaag gacttgttcg agcaaaaggc cgctcaactt gctacagaga tcgccgacat
     5221 caagtcaaag tacgacgagg agaagagtct tcgagccgct gccgagcaaa aggtaactca
     5281 ccttactgag gacctcaata agcaaactac tgtcatccaa gacctcaaga cggagttgct
     5341 ccaacggcct gggatagagg acgtagctgt tcttaagaag gagttggttc aagttcaaac
     5401 gttgatggac aatatgactt tggagcgaga gagggagtca gagaagttga aggacgagtg
     5461 taagaagttg caatccgagc acgcccacct cgaggccact ataaatcaat tgagaagtga
     5521 gctcgccaag ggacctcaag aggtagcagt ctacgtccaa gagatccaaa agctcaaggg
     5581 tagtatcaat gagcttacac aaaagaatca aaatctcact gagaagctcc aaaagaagga
     5641 cttggactac acgcacctag aggagaagca caatgaggag tccgcaagta ggaagacttt
     5701 gcaagcaagt cttcaccaaa gggacctcga ctgtcaacaa ttgcaagccc gactcacggc
     5761 atcagagtca tctctacaac gagcacaagg agagttgtcc gagaaggccg aggccgcaca
     5821 aaagctccga gaggagctca gggaagtcga gtcaactcga caacacttga aggttgaggt
     5881 aaagcaacta caacaacaaa gagaggagaa ggagcaacac ggattgcaat tgcaaggcga
     5941 ggtctcacaa ctccactgta agctactaga gacagagagg caactagggg aagcccacgg
     6001 gcgattgaag gagcaaaggc aactcagttc tgagaagctc atggagaagg agcaacaagt
     6061 cgcagacctt caattgaagc tctctaggtt ggaagagcaa ttgaaggaga aggtcacgaa
     6121 ttccacggag ctacaacacc aactcgagaa gtctaagcaa caacaccaag agcaacaagc
     6181 acttcaacaa tccgctactg ctaagcttag ggaagcacaa aatgacctcg agcaagttct
     6241 caggcaaata ggggacaagg accaaaagat ccaaaatctt gaggctctcc ttcaaaaggg
     6301 taaggaaagt gttagtctcc tcgagaagga gcgagaggac ttgtacgcaa agatccaagc
     6361 aggagaggga gagactgccg tcttgaatca attgcaagag aagaatcacg cacttcaaca
     6421 acaactcact caactaacag agaagttgaa gaatcaatct gagtcccaca agcaagccga
     6481 ggagaatctt cacgaccaag tccaagagca aaaggcccac ttgcgagcag cacaagacag
     6541 ggttttgtcc ttggagacgt cagtcagtga gttgtcttct caactcaatg agtccaagga
     6601 g
//
