LOCUS       sequence-168836-        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   Kim E, Ilic N, Shrestha Y, Zou L, Kamburov A, Zhu C, Yang X, Lubonja
            R, Tran N, Nguyen C, Lawrence MS, Piccioni F, Bagul M, Doench JG, 
            Chouinard CR, Wu X, Hogstrom L, Natoli T, Tamayo P, Horn H, Corsello
            SM, Lage K, Root DE, Subramanian A, Golub TR, Getz G, Boehm JS, Hahn
            WC
  TITLE     Systematic Functional Interrogation of Rare Cancer Variants 
            Identifies Oncogenic Alleles.
  JOURNAL   Cancer Discov. 2016 Jul;6(7):714-26. doi: 
            10.1158/2159-8290.CD-16-0160. Epub 2016 May 4.
  PUBMED    27147599
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: 
            "10.1158/2159-8290.CD-16-0160"; journalName: "Cancer Discov"; date: 
            "2016-07"; volume: "6"; issue: "7"; pages: "714-26"
COMMENT     Sequence Label: pDONR223_KEAP1_WT
FEATURES             Location/Qualifiers
     source          1..4668
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     382..401
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     635..652
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(811..838)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(930..1016)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1065..1087
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1079..1096
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1080..1096
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1112..1211
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3090..3189)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3206..3225)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3207..3225)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3230..3246)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3230..3246)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3677..4468
                     /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(4561..4668,1..481)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
       61 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
      121 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
      181 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
      241 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
      301 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
      361 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
      421 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
      481 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
      541 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
      601 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
      661 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
      721 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac
      781 cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc
      841 ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc
      901 ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac
      961 aaacaacaga taaaacgaaa ggcccagtct tccgactgag cctttcgttt tatttgatgc
     1021 ctggcagttc cctactctcg cgttaacgct agcatggatg ttttcccagt cacgacgttg
     1081 taaaacgacg gccagtctta agctcgggcc ccaaataatg attttatttt gactgatagt
     1141 gacctgttcg ttgcaacaaa ttgatgagca atgctttttt ataatgccaa ctttgtacaa
     1201 aaaagttggc accatgcagc cagatcccag gcctagcggg gctggggcct gctgccgatt
     1261 cctgcccctg cagtcacagt gccctgaggg ggcaggggac gcggtgatgt acgcctccac
     1321 tgagtgcaag gcggaggtga cgccctccca gcatggcaac cgcaccttca gctacaccct
     1381 ggaggatcat accaagcagg cctttggcat catgaacgag ctgcggctca gccagcagct
     1441 gtgtgacgtc acactgcagg tcaagtacca ggatgcaccg gccgcccagt tcatggccca
     1501 caaggtggtg ctggcctcat ccagccctgt cttcaaggcc atgttcacca acgggctgcg
     1561 ggagcagggc atggaggtgg tgtccattga gggtatccac cccaaggtca tggagcgcct
     1621 cattgaattc gcctacacgg cctccatctc catgggcgag aagtgtgtcc tccacgtcat
     1681 gaacggtgct gtcatgtacc agatcgacag cgttgtccgt gcctgcagtg acttcctggt
     1741 gcagcagctg gaccccagca atgccatcgg catcgccaac ttcgctgagc agattggctg
     1801 tgtggagttg caccagcgtg cccgggagta catctacatg cattttgggg aggtggccaa
     1861 gcaagaggag ttcttcaacc tgtcccactg ccaactggtg accctcatca gccgggacga
     1921 cctgaacgtg cgctgcgagt ccgaggtctt ccacgcctgc atcaactggg tcaagtacga
     1981 ctgcgaacag cgacggttct acgtccaggc gctgctgcgg gccgtgcgct gccactcgtt
     2041 gacgccgaac ttcctgcaga tgcagctgca gaagtgcgag atcctgcagt ccgactcccg
     2101 ctgcaaggac tacctggtca agatcttcga ggagctcacc ctgcacaagc ccacgcaggt
     2161 gatgccctgc cgggcgccca aggtgggccg cctgatctac accgcgggcg gctacttccg
     2221 acagtcgctc agctacctgg aggcttacaa ccccagtgac ggcacctggc tccggttggc
     2281 ggacctgcag gtgccgcgga gcggcctggc cggctgcgtg gtgggcgggc tgttgtacgc
     2341 cgtgggcggc aggaacaact cgcccgacgg caacaccgac tccagcgccc tggactgtta
     2401 caaccccatg accaatcagt ggtcgccctg cgcccccatg agcgtgcccc gtaaccgcat
     2461 cggggtgggg gtcatcgatg gccacatcta tgccgtcggc ggctcccacg gctgcatcca
     2521 ccacaacagt gtggagaggt atgagccaga gcgggatgag tggcacttgg tggccccaat
     2581 gctgacacga aggatcgggg tgggcgtggc tgtcctcaat cgtctccttt atgccgtggg
     2641 gggctttgac gggacaaacc gccttaattc agctgagtgt tactacccag agaggaacga
     2701 gtggcgaatg atcacagcaa tgaacaccat ccgaagcggg gcaggcgtct gcgtcctgca
     2761 caactgtatc tatgctgctg ggggctatga tggtcaggac cagctgaaca gcgtggagcg
     2821 ctacgatgtg gaaacagaga cgtggacttt cgtagccccc atgaagcacc ggcgaagtgc
     2881 cctggggatc actgtccacc aggggagaat ctacgtcctt ggaggctatg atggtcacac
     2941 gttcctggac agtgtggagt gttacgaccc agatacagac acctggagcg aggtgacccg
     3001 aatgacatcg ggccggagtg gggtgggcgt ggctgtcacc atggagccct gccggaagca
     3061 gattgaccag cagaactgta ccagttgatt gccaactttc ttgtacaaag ttggcattat
     3121 aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat
     3181 cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt
     3241 ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa
     3301 tatatcatca tgcctcctct agaccagcca ggacagaaat gcctcgactt cgctgctacc
     3361 caaggttgcc gggtgacgca caccgtggaa acggatgaag gcacgaaccc agtggacata
     3421 agcctgttcg gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca actggtccag
     3481 aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg cttgttatga
     3541 ctgttttttt ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt
     3601 gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagggca gtcgccctaa
     3661 aacaaagtta aacattatga gggaagcggt gatcgccgaa gtatcgactc aactatcaga
     3721 ggtagttggc gtcatcgagc gccatctcga accgacgttg ctggccgtac atttgtacgg
     3781 ctccgcagtg gatggcggcc tgaagccaca cagtgatatt gatttgctgg ttacggtgac
     3841 cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg aaacttcggc
     3901 ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc accattgttg tgcacgacga
     3961 catcattccg tggcgttatc cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa
     4021 tgacattctt gcaggtatct tcgagccagc cacgatcgac attgatctgg ctatcttgct
     4081 gacaaaagca agagaacata gcgttgcctt ggtaggtcca gcggcggagg aactctttga
     4141 tccggttcct gaacaggatc tatttgaggc gctaaatgaa accttaacgc tatggaactc
     4201 gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc gcatttggta
     4261 cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg caatggagcg
     4321 cctgccggcc cagtatcagc ccgtcatact tgaagctaga caggcttatc ttggacaaga
     4381 agaagatcgc ttggcctcgc gcgcagatca gttggaagaa tttgtccact acgtgaaagg
     4441 cgagatcacc aaggtagtcg gcaaataacc ctcgagccac ccatgaccaa aatcccttaa
     4501 cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4561 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4621 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaa
//
