LOCUS       sequence-163907-        5031 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing CHUK , part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5031)
  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 5031)
  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_CHUK_WT
FEATURES             Location/Qualifiers
     source          1..5031
                     /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(2910..3009)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3026..3045)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3027..3045)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3050..3066)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3050..3066)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3497..4288
                     /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      4381..4969
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4870..4889
                     /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 atggagcggc ccccggggct gcggccgggc gcgggcgggc cctgggagat
      721 gcgggagcgg ctgggcaccg gcggcttcgg gaacgtctgt ctgtaccagc atcgggaact
      781 tgatctcaaa atagcaatta agtcttgtcg cctagagcta agtaccaaaa acagagaacg
      841 atggtgccat gaaatccaga ttatgaagaa gttgaaccat gccaatgttg taaaggcctg
      901 tgatgttcct gaagaattga atattttgat tcatgatgtg cctcttctag caatggaata
      961 ctgttctgga ggagatctcc gaaagctgct caacaaacca gaaaattgtt gtggacttaa
     1021 agaaagccag atactttctt tactaagtga tatagggtct gggattcgat atttgcatga
     1081 aaacaaaatt atacatcgag atctaaaacc tgaaaacata gttcttcagg atgttggtgg
     1141 aaagataata cataaaataa ttgatctggg atatgccaaa gatgttgatc aaggaagtct
     1201 gtgtacatct tttgtgggaa cactgcagta tctggcccca gagctctttg agaataagcc
     1261 ttacacagcc actgttgatt attggagctt tgggaccatg gtatttgaat gtattgctgg
     1321 atataggcct tttttgcatc atctgcagcc atttacctgg catgagaaga ttaagaagaa
     1381 ggatccaaag tgtatatttg catgtgaaga gatgtcagga gaagttcggt ttagtagcca
     1441 tttacctcaa ccaaatagcc tttgtagttt aatagtagaa cccatggaaa actggctaca
     1501 gttgatgttg aattgggacc ctcagcagag aggaggacct gttgacctta ctttgaagca
     1561 gccaagatgt tttgtattaa tggatcacat tttgaatttg aagatagtac acatcctaaa
     1621 tatgacttct gcaaagataa tttcttttct gttaccacct gatgaaagtc ttcattcact
     1681 acagtctcgt attgagcgtg aaactggaat aaatactggt tctcaagaac ttctttcaga
     1741 gacaggaatt tctctggatc ctcggaaacc agcctctcaa tgtgttctag atggagttag
     1801 aggctgtgat agctatatgg tttatttgtt tgataaaagt aaaactgtat atgaagggcc
     1861 atttgcttcc agaagtttat ctgattgtgt aaattatatt gtacaggaca gcaaaataca
     1921 gcttccaatt atacagctgc gtaaagtgtg ggctgaagca gtgcactatg tgtctggact
     1981 aaaagaagac tatagcaggc tctttcaggg acaaagggca gcaatgttaa gtcttcttag
     2041 atataatgct aacttaacaa aaatgaagaa cactttgatc tcagcatcac aacaactgaa
     2101 agctaaattg gagttttttc acaaaagcat tcagcttgac ttggagagat acagcgagca
     2161 gatgacgtat gggatatctt cagaaaaaat gctaaaagca tggaaagaaa tggaagaaaa
     2221 ggccatccac tatgctgagg ttggtgtcat tggatacctg gaggatcaga ttatgtcttt
     2281 gcatgctgaa atcatggagc tacagaagag cccctatgga agacgtcagg gagacttgat
     2341 ggaatctctg gaacagcgtg ccattgatct atataagcag ttaaaacaca gaccttcaga
     2401 tcactcctac agtgacagca cagagatggt gaaaatcatt gtgcacactg tgcagagtca
     2461 ggaccgtgtg ctcaaggagc tgtttggtca tttgagcaag ttgttgggct gtaagcagaa
     2521 gattattgat ctactcccta aggtggaagt ggccctcagt aatatcaaag aagctgacaa
     2581 tactgtcatg ttcatgcagg gaaaaaggca gaaagaaata tggcatctcc ttaaaattgc
     2641 ctgtacacag agttctgccc ggtcccttgt aggatccagt ctagaaggtg cagtaacccc
     2701 tcagacatca gcatggctgc ccccgacttc agcagaacat gatcattctc tgtcatgtgt
     2761 ggtaactcct caagatgggg agacttcagc acaaatgata gaagaaaatt tgaactgcct
     2821 tggccattta agcactatta ttcatgaggc aaatgaggaa cagggcaata gtatgatgaa
     2881 tcttgattgg agttggttaa cagaatgatt gccaactttc ttgtacaaag ttggcattat
     2941 aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat
     3001 cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg tcatagctgt
     3061 ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac aagataaaaa
     3121 tatatcatca tgcctcctct agaccagcca ggacagaaat gcctcgactt cgctgctacc
     3181 caaggttgcc gggtgacgca caccgtggaa acggatgaag gcacgaaccc agtggacata
     3241 agcctgttcg gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca actggtccag
     3301 aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg cttgttatga
     3361 ctgttttttt ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt
     3421 gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagggca gtcgccctaa
     3481 aacaaagtta aacattatga gggaagcggt gatcgccgaa gtatcgactc aactatcaga
     3541 ggtagttggc gtcatcgagc gccatctcga accgacgttg ctggccgtac atttgtacgg
     3601 ctccgcagtg gatggcggcc tgaagccaca cagtgatatt gatttgctgg ttacggtgac
     3661 cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg aaacttcggc
     3721 ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc accattgttg tgcacgacga
     3781 catcattccg tggcgttatc cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa
     3841 tgacattctt gcaggtatct tcgagccagc cacgatcgac attgatctgg ctatcttgct
     3901 gacaaaagca agagaacata gcgttgcctt ggtaggtcca gcggcggagg aactctttga
     3961 tccggttcct gaacaggatc tatttgaggc gctaaatgaa accttaacgc tatggaactc
     4021 gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc gcatttggta
     4081 cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg caatggagcg
     4141 cctgccggcc cagtatcagc ccgtcatact tgaagctaga caggcttatc ttggacaaga
     4201 agaagatcgc ttggcctcgc gcgcagatca gttggaagaa tttgtccact acgtgaaagg
     4261 cgagatcacc aaggtagtcg gcaaataacc ctcgagccac ccatgaccaa aatcccttaa
     4321 cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4381 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4441 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4501 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     4561 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4621 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4681 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4741 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4801 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4861 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4921 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4981 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt t
//
