LOCUS       sequence-161636-        4668 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing KEAP1, part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4668)
  AUTHORS   Berger AH, Brooks AN, Wu X, Shrestha Y, Chouinard C, Piccioni F, 
            Bagul M, Kamburov A, Imielinski M, Hogstrom L, Zhu C, Yang X, Pantel
            S, Sakai R, Watson J, Kaplan N, Campbell JD, Singh S, Root DE, 
            Narayan R, Natoli T, Lahr DL, Tirosh I, Tamayo P, Getz G, Wong B, 
            Doench J, Subramanian A, Golub TR, Meyerson M, Boehm JS
  TITLE     High-throughput Phenotyping of Lung Cancer Somatic Mutations.
  JOURNAL   Cancer Cell. 2016 Aug 8;30(2):214-28. doi: 
            10.1016/j.ccell.2016.06.022. Epub 2016 Jul 28.
  PUBMED    27478040
REFERENCE   2  (bases 1 to 4668)
  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"; doi: 
            "10.1016/j.ccell.2016.06.022"; journalName: "Cancer Cell"; date: 
            "2016-08-8- 8"; volume: "30"; issue: "2"; pages: "214-28"
COMMENT     Sequence Label: pDONR223_KEAP1_p.L268P
FEATURES             Location/Qualifiers
     source          1..4668
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     180..197
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(356..383)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(475..561)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     610..632
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     624..641
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     625..641
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    657..756
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2635..2734)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2751..2770)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2752..2770)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2775..2791)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2775..2791)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3222..4013
                     /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"
     rep_origin      join(4106..4668,1..26)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4595..4614
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
       61 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
      121 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
      181 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
      241 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
      301 agcgcaacgc aattaatacg cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag
      361 gccatccgtc aggatggcct tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc
      421 tgcccgccac cctccgggcc gttgcttcac aacgttcaaa tccgctcccg gcggatttgt
      481 cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtcttccga
      541 ctgagccttt cgttttattt gatgcctggc agttccctac tctcgcgtta acgctagcat
      601 ggatgttttc ccagtcacga cgttgtaaaa cgacggccag tcttaagctc gggccccaaa
      661 taatgatttt attttgactg atagtgacct gttcgttgca acaaattgat gagcaatgct
      721 tttttataat gccaactttg tacaaaaaag ttggcaccat gcagccagat cccaggccta
      781 gcggggctgg ggcctgctgc cgattcctgc ccctgcagtc acagtgccct gagggggcag
      841 gggacgcggt gatgtacgcc tccactgagt gcaaggcgga ggtgacgccc tcccagcatg
      901 gcaaccgcac cttcagctac accctggagg atcataccaa gcaggccttt ggcatcatga
      961 acgagctgcg gctcagccag cagctgtgtg acgtcacact gcaggtcaag taccaggatg
     1021 caccggccgc ccagttcatg gcccacaagg tggtgctggc ctcatccagc cctgtcttca
     1081 aggccatgtt caccaacggg ctgcgggagc agggcatgga ggtggtgtcc attgagggta
     1141 tccaccccaa ggtcatggag cgcctcattg aattcgccta cacggcctcc atctccatgg
     1201 gcgagaagtg tgtcctccac gtcatgaacg gtgctgtcat gtaccagatc gacagcgttg
     1261 tccgtgcctg cagtgacttc ctggtgcagc agctggaccc cagcaatgcc atcggcatcg
     1321 ccaacttcgc tgagcagatt ggctgtgtgg agttgcacca gcgtgcccgg gagtacatct
     1381 acatgcattt tggggaggtg gccaagcaag aggagttctt caacctgtcc cactgccaac
     1441 tggtgaccct catcagccgg gacgacctga acgtgcgctg cgagtccgag gtcttccacg
     1501 cctgcatcaa ctgggtcaag tacgactgcg aacagcgacg gttctacgtc caggcgctgc
     1561 cgcgggccgt gcgctgccac tcgttgacgc cgaacttcct gcagatgcag ctgcagaagt
     1621 gcgagatcct gcagtccgac tcccgctgca aggactacct ggtcaagatc ttcgaggagc
     1681 tcaccctgca caagcccacg caggtgatgc cctgccgggc gcccaaggtg ggccgcctga
     1741 tctacaccgc gggcggctac ttccgacagt cgctcagcta cctggaggct tacaacccca
     1801 gtgacggcac ctggctccgg ttggcggacc tgcaggtgcc gcggagcggc ctggccggct
     1861 gcgtggtggg cgggctgttg tacgccgtgg gcggcaggaa caactcgccc gacggcaaca
     1921 ccgactccag cgccctggac tgttacaacc ccatgaccaa tcagtggtcg ccctgcgccc
     1981 ccatgagcgt gccccgtaac cgcatcgggg tgggggtcat cgatggccac atctatgccg
     2041 tcggcggctc ccacggctgc atccaccaca acagtgtgga gaggtatgag ccagagcggg
     2101 atgagtggca cttggtggcc ccaatgctga cacgaaggat cggggtgggc gtggctgtcc
     2161 tcaatcgtct cctttatgcc gtggggggct ttgacgggac aaaccgcctt aattcagctg
     2221 agtgttacta cccagagagg aacgagtggc gaatgatcac agcaatgaac accatccgaa
     2281 gcggggcagg cgtctgcgtc ctgcacaact gtatctatgc tgctgggggc tatgatggtc
     2341 aggaccagct gaacagcgtg gagcgctacg atgtggaaac agagacgtgg actttcgtag
     2401 cccccatgaa gcaccggcga agtgccctgg ggatcactgt ccaccagggg agaatctacg
     2461 tccttggagg ctatgatggt cacacgttcc tggacagtgt ggagtgttac gacccagata
     2521 cagacacctg gagcgaggtg acccgaatga catcgggccg gagtggggtg ggcgtggctg
     2581 tcaccatgga gccctgccgg aagcagattg accagcagaa ctgtaccagt tgattgccaa
     2641 ctttcttgta caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac
     2701 aggtcactat cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg
     2761 agtcgtatta catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct
     2821 gatgttacat tgcacaagat aaaaatatat catcatgcct cctctagacc agccaggaca
     2881 gaaatgcctc gacttcgctg ctacccaagg ttgccgggtg acgcacaccg tggaaacgga
     2941 tgaaggcacg aacccagtgg acataagcct gttcggttcg taagctgtaa tgcaagtagc
     3001 gtatgcgctc acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag
     3061 tggcggtttt catggcttgt tatgactgtt tttttggggt acagtctatg cctcgggcat
     3121 ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc agcaacgatg
     3181 ttacgcagca gggcagtcgc cctaaaacaa agttaaacat tatgagggaa gcggtgatcg
     3241 ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga
     3301 cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag ccacacagtg
     3361 atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga
     3421 tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc cgcgctgtag
     3481 aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct aagcgcgaac
     3541 tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag ccagccacga
     3601 tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt gccttggtag
     3661 gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt gaggcgctaa
     3721 atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggcgatgag cgaaatgtag
     3781 tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg
     3841 tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc atacttgaag
     3901 ctagacaggc ttatcttgga caagaagaag atcgcttggc ctcgcgcgca gatcagttgg
     3961 aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa taaccctcga
     4021 gccacccatg accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc
     4081 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4141 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4201 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4261 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4321 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     4381 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     4441 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     4501 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4561 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4621 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgct
//
