LOCUS       sequence-168684-        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_p.T142M
FEATURES             Location/Qualifiers
     source          1..4668
                     /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(2547..2646)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2663..2682)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2664..2682)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2687..2703)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2687..2703)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3134..3925
                     /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      4018..4606
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4507..4526
                     /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 agttggcacc atgcagccag atcccaggcc tagcggggct ggggcctgct gccgattcct
      721 gcccctgcag tcacagtgcc ctgagggggc aggggacgcg gtgatgtacg cctccactga
      781 gtgcaaggcg gaggtgacgc cctcccagca tggcaaccgc accttcagct acaccctgga
      841 ggatcatacc aagcaggcct ttggcatcat gaacgagctg cggctcagcc agcagctgtg
      901 tgacgtcaca ctgcaggtca agtaccagga tgcaccggcc gcccagttca tggcccacaa
      961 ggtggtgctg gcctcatcca gccctgtctt caaggccatg ttcaccaacg ggctgcggga
     1021 gcagggcatg gaggtggtgt ccattgaggg tatccacccc aaggtcatgg agcgcctcat
     1081 tgaattcgcc tacatggcct ccatctccat gggcgagaag tgtgtcctcc acgtcatgaa
     1141 cggtgctgtc atgtaccaga tcgacagcgt tgtccgtgcc tgcagtgact tcctggtgca
     1201 gcagctggac cccagcaatg ccatcggcat cgccaacttc gctgagcaga ttggctgtgt
     1261 ggagttgcac cagcgtgccc gggagtacat ctacatgcat tttggggagg tggccaagca
     1321 agaggagttc ttcaacctgt cccactgcca actggtgacc ctcatcagcc gggacgacct
     1381 gaacgtgcgc ttcgagtccg aggtcttcca cgcctgcatc aactgggtca agtacgactg
     1441 cgaacagcga cggttctacg tccaggcgct gctgcgggcc gtgcgctgcc actcgttgac
     1501 gccgaacttc ctgcagatgc agctgcagaa gtgcgagatc ctgcagtccg actcccgctg
     1561 caaggactac ctggtcaaga tcttcgagga gctcaccctg cacaagccca cgcaggtgat
     1621 gccctgccgg gcgcccaagg tgggccgcct gatctacacc gcgggcggct acttccgaca
     1681 gtcgctcagc tacctggagg cttacaaccc cagtgacggc acctggctcc ggttggcgga
     1741 cctgcaggtg ccgcggagcg gcctggccgg ctgcgtggtg ggcgggctgt tgtacgccgt
     1801 gggcggcagg aacaactcgc ccgacggcaa caccgactcc agcgccctgg actgttacaa
     1861 ccccatgacc aatcagtggt cgccctgcgc ccccatgagc gtgccccgta accgcatcgg
     1921 ggtgggggtc atcgatggcc acatctatgc cgtcggcggc tcccacggct gcatccacca
     1981 caacagtgtg gagaggtatg agccagagcg ggatgagtgg cacttggtgg ccccaatgct
     2041 gacacgaagg atcggggtgg gcgtggctgt cctcaatcgt ctcctttatg ccgtgggggg
     2101 ctttgacggg acaaaccgcc ttaattcagc tgagtgttac tacccagaga ggaacgagtg
     2161 gcgaatgatc acagcaatga acaccatccg aagcggggca ggcgtctgcg tcctgcacaa
     2221 ctgtatctat gctgctgggg gctatgatgg tcaggaccag ctgaacagcg tggagcgcta
     2281 cgatgtggaa acagagacgt ggactttcgt agcccccatg aagcaccggc gaagtgccct
     2341 ggggatcact gtccaccagg ggagaatcta cgtccttgga ggctatgatg gtcacacgtt
     2401 cctggacagt gtggagtgtt acgacccaga tacagacacc tggagcgagg tgacccgaat
     2461 gacatcgggc cggagtgggg tgggcgtggc tgtcaccatg gagccctgcc ggaagcagat
     2521 tgaccagcag aactgtacca gttgattgcc aactttcttg tacaaagttg gcattataag
     2581 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
     2641 tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc
     2701 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     2761 atcatcatgc ctcctctaga ccagccagga cagaaatgcc tcgacttcgc tgctacccaa
     2821 ggttgccggg tgacgcacac cgtggaaacg gatgaaggca cgaacccagt ggacataagc
     2881 ctgttcggtt cgtaagctgt aatgcaagta gcgtatgcgc tcacgcaact ggtccagaac
     2941 cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt gttatgactg
     3001 tttttttggg gtacagtcta tgcctcgggc atccaagcag caagcgcgtt acgccgtggg
     3061 tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagggcagtc gccctaaaac
     3121 aaagttaaac attatgaggg aagcggtgat cgccgaagta tcgactcaac tatcagaggt
     3181 agttggcgtc atcgagcgcc atctcgaacc gacgttgctg gccgtacatt tgtacggctc
     3241 cgcagtggat ggcggcctga agccacacag tgatattgat ttgctggtta cggtgaccgt
     3301 aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa cttcggcttc
     3361 ccctggagag agcgagattc tccgcgctgt agaagtcacc attgttgtgc acgacgacat
     3421 cattccgtgg cgttatccag ctaagcgcga actgcaattt ggagaatggc agcgcaatga
     3481 cattcttgca ggtatcttcg agccagccac gatcgacatt gatctggcta tcttgctgac
     3541 aaaagcaaga gaacatagcg ttgccttggt aggtccagcg gcggaggaac tctttgatcc
     3601 ggttcctgaa caggatctat ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc
     3661 gcccgactgg gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca tttggtacag
     3721 cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa tggagcgcct
     3781 gccggcccag tatcagcccg tcatacttga agctagacag gcttatcttg gacaagaaga
     3841 agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gtccactacg tgaaaggcga
     3901 gatcaccaag gtagtcggca aataaccctc gagccaccca tgaccaaaat cccttaacgt
     3961 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4021 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4081 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4141 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4201 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4261 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4321 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4381 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4441 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4501 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4561 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4621 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtt
//
