LOCUS       sequence-168693-        8025 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing WNK3, part of the Target Accelerator
            Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8025)
  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 8025)
  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_WNK3_p.E417Q
FEATURES             Location/Qualifiers
     source          1..8025
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      20..608
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     509..528
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     762..779
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(938..965)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1057..1143)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1192..1214
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1206..1223
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1207..1223
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1239..1338
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(6574..6673)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(6690..6709)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6691..6709)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6714..6730)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6714..6730)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             7161..7952
                     /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"
ORIGIN
        1 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
       61 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
      121 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
      181 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
      241 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
      301 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
      361 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
      421 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
      481 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
      541 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
      601 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
      661 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
      721 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
      781 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
      841 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata
      901 cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc
      961 cttctgctta gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg
     1021 ccgttgcttc acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt
     1081 caccgacaaa caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat
     1141 ttgatgcctg gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac
     1201 gacgttgtaa aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac
     1261 tgatagtgac ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt
     1321 tgtacaaaaa agttggcacc atggccactg attcagggga tccagccagc acagaagatt
     1381 ctgagaaacc tgatggaatt tcatttgaaa acagagttcc ccaggtcgct gcaactttga
     1441 cagtagaagc tagactaaag gagaaaaaca gtaccttctc tgcttctggg gaaactgtag
     1501 aaaggaagag atttttccga aagagtgttg aaatgacgga agatgacaaa gttgccgaat
     1561 catcccccaa agatgagaga attaaggctg caatgaatat tccaagagta gataagcttc
     1621 cttcaaatgt gttgagaggt ggacaagaag ttaaatatga acagtgttca aagtcaacct
     1681 cagaaatctc aaaagattgt ttcaaggaga aaaatgaaaa ggaaatggaa gaagaagcag
     1741 aaatgaaggc tgtagctact tctcctagtg gcagattcct gaaatttgac atagaactag
     1801 gaagaggagc atttaaaaca gtatataaag gactggacac tgaaacatgg gttgaggttg
     1861 cttggtgtga gctgcaggac cgaaagttaa ccaaagctga gcagcaaaga ttcaaggaag
     1921 aagcagagat gttgaagggt ctccagcacc ccaatatagt tcgattttat gattcctggg
     1981 aatctatatt aaaaggaaag aaatgtattg tattagtgac tgaactaatg acatctggga
     2041 ccttaaagac gtacttaaaa cgatttaaag tcatgaaacc aaaggtctta aggagctggt
     2101 gcaggcaaat tttaaagggg ttgcagttct tgcacactag gactcctcct attattcacc
     2161 gggatctgaa gtgtgacaat attttcatca cgggacccac tggatctgtg aagattggtg
     2221 atctaggatt agccacctta atgcgtacct catttgctaa gagtgtcatt ggaactcctg
     2281 agtttatggc tccagagatg tatgaagaac actatgatga atccgtagat gtttatgctt
     2341 ttggaatgtg tatgctggaa atggccacat cggagtatcc ttattctgag tgtcagaatg
     2401 cagctcaaat ataccggaaa gtaactagtg gcataaaacc agccagcttc aataaagtca
     2461 ctgatcctga agtcaaagaa atcattgaag gatgtattcg tcaaaacaaa tctgaaaggt
     2521 tgtctatcag ggacctatta aaccatgcat tttttgctga ggatacagga ctgagggtgg
     2581 agttagcaca agaagatgat tgctcaaatt catcccttgc tttaagactc tgggttgaag
     2641 accctaaaaa attgaaaggc aaacacaaag acaatgaagc tattgaattt agtttcaact
     2701 tagaaacaga tacacctgag gaagtagcat atgaaatggt caagtctggg ttcttccatg
     2761 aaagtgattc caaagctgtt gctaaatcca ttagagaccg ggtgacgcca ataaagaaga
     2821 caagagagaa gaagcctgct ggctgtttgg aagaacgcag ggattctcag tgcaagtcta
     2881 tggggaatgt attccctcag ccccagaata caactttacc ccttgctccc gctcagcaaa
     2941 ctggggctga atgtgaagaa actgaagttg atcaacatgt tagacaacag cttctacaaa
     3001 gaaaaccaca gcagcactgc tcctctgtta caggtgacaa tttgtctgag gcaggagctg
     3061 catcagttat acattcagat acttcaagtc agcccagtgt agcctattcc tcaaatcaaa
     3121 cgatgggctc tcaaatggtt tctaatatcc cgcaggctga agtaaatgtt ccagggcaaa
     3181 tttattcatc tcagcaacta gtaggacatt accagcaagt ttcagggtta cagaagcatt
     3241 caaagctgac tcagccgcag attttgcctt tggttcaagg tcagtccact gttttacctg
     3301 tacatgtcct tggaccgaca gttgtttcac aaccccaggt ttccccatta actgttcaga
     3361 aggtcccaca gataaagcct gtatcccaac cagttggagc tgaacaacaa gcagctcttc
     3421 taaaaccaga tttagttcga agcttgaatc aagatgtggc aactacgaag gaaaacgtca
     3481 gtagccctga taacccaagt ggaaatggca aacaggatcg gatcaaacag agaagagctt
     3541 cctgtccccg accagagaag gggactaaat ttcagcttac cgtccttcag gtatcaacct
     3601 ctggagataa catggtagag tgtcagctgg agacacacaa caacaagatg gtcaccttca
     3661 agtttgatgt tgatggtgat gcgccagagg atattgcaga ctatatggtt gaagataact
     3721 ttgtgctgga aagtgagaaa gaaaaatttg tagaagaatt gagagctatt gtaggtcaag
     3781 cccaggagat ccttcatgtc cactttgcca cagaaagagc cactggagtt gattctatta
     3841 ctgtggactc caacagtagc cagacagggt catctgaaca agtacagata aattctacat
     3901 ctactcaaac cagcaatgaa tctgctcctc agtcatcccc agttggtcgg tggcgattct
     3961 gtatcaatca gacgataaga aaccgtgaga ctcagtctcc tccttctctt cagcattcca
     4021 tgtctgcggt tcctggccga catccacttc ctagtccaaa aaacacaagt aataaggaaa
     4081 tatcacggga cacattgctc actatagaaa ataatccatg ccaccgtgca cttttcacct
     4141 ccaaatcaga acacaaggat gtggttgatg gtaagatttc tgaatgtgct agtgtagaaa
     4201 ccaagcagcc agctatactt tatcaagtgg aagataacag gcagataatg gcaccagtta
     4261 ctaatagttc cagttactct actacttcag ttcgtgcagt tccagctgaa tgtgagggac
     4321 tcaccaaaca agcaagcata ttcatacctg tgtatccatg tcaccaaact gccagtcagg
     4381 ctgatgcact tatgtcccat cctggcgaat caactcagac ttctggtaac tctcttacaa
     4441 ctctggcatt tgatcaaaag cctcaaacct tatcagtaca gcagccagct atggatgcag
     4501 agtttatttc tcaagaagga gaaactacag tgaacactga agcaagttct cctaagacag
     4561 tcattcccac tcagacccct ggccttgaac caactaccct tcaacccact actgtcctgg
     4621 aatcagatgg agaaagacct ccaaaactgg agtttgcaga caaccgaatt aaaactctgg
     4681 atgaaaaatt aagaaacttg ctctatcagg agcacagcat ctctagcatc tatcccgaga
     4741 gtcagaagga tacccaaagc atagactctc cattttcttc ctctgctgaa gataccctct
     4801 cctgtccagt gacagaagtc atagccatca gtcactgtgg aattaaagat agccctgtac
     4861 aatcccctaa tttccaacag acaggctcta agcttctgtc caatgtggct gcaagtcagc
     4921 ctgctaatat atcagtgttc aaaagggacc tgaatgtgat aacttctgta cccagcgaat
     4981 tgtgtttaca tgagatgtcc tcagatgctt cacttccagg ggatccagag gcctatcctg
     5041 ctgctgtgtc aagcggtgga gccattcatc tgcagacagg agtggaaaca gaagagatga
     5101 gatcagcaat tgctcctgat cccattcctc tgacacggga gtccacagct gatactaggg
     5161 ctttgaatag atgtaaagcg atgagtggat catttcagcg gggtcggttc caggtgatta
     5221 caattcctca gcagcagtca gcaaaaatga catcttttgg aatagaacac atatcagtgt
     5281 tcagtgagac aaaccattct agtgaagaag cctttattaa aacagcaaag tctcagttgg
     5341 tagaaataga acctgccaca caaaatccaa aaacttcgtt ttcttatgag aagttacaag
     5401 ctcttcagga aacctgtaaa gaaaataaag gagttcccaa acaaggtgac aacttcttat
     5461 ctttcagcgc agcttgtgag actgatgtat cttcagtgac cccagaaaag gaatttgaag
     5521 aaacttcagc cacaggaagt agcatgcagt ctggatctga actgttgctt aaagagagag
     5581 agatattgac tgctgggaaa cagcctagct ctgatagtga attttcagcc agtcttgctg
     5641 gcagtggaaa gtcagtggca aagactggtc cagagagtaa tcagtgctta ccacaccacg
     5701 aagaacaagc ttatgctcaa acacagagtt cactcttcta ttcgccatct tccccaatga
     5761 gcagtgatga tgaatcagaa atagaggatg aggacttgaa ggtggagctt caaagattac
     5821 gagaaaaaca cattcaggag gtggtaaatc ttcaaaccca gcagaataag gagctgcagg
     5881 agctctatga acgccttcgg tcaattaaag atagcaaaac ccaatctact gagattcctt
     5941 tgccacctgc atcaccacgt cgaccaagat ctttcaaaag caaacttcga agccgccccc
     6001 agtccttgac acatgtggac aatggcatag ttgctacaga tccactgtgt gtggagagta
     6061 atgcagcatc atgccaacag tctccagcca gtaaaaaagg gatgttcaca gatgacttac
     6121 acaagctggt ggatgactgg acaaaggaag cagtaggaaa ttctcttatt aagccaagtt
     6181 taaaccaact taaacaaagt caacacaaac tagagacaga aaactggaat aaagtatctg
     6241 aaaatactcc gtctactatg ggctacacat caacatggat ttcttctctg tcccaaatcc
     6301 gtggagctgt cccaacttcc ttgccacaag gactctcact cccttcattt cctgggccat
     6361 tatcatcata tggaatgcct cacgtttgtc agtataatgc tgtggcgggg gcggggtatc
     6421 cagtacagtg ggtaggaatt tcaggaacaa cacaacaatc tgtagtaatt cccgcccaat
     6481 ctgggggacc attccagcca gggatgaata tgcaggcatt tccaacttca tcagtgcaga
     6541 atcctgccac aatccctcct ggtcctaaat gattgccaac tttcttgtac aaagttggca
     6601 ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata
     6661 aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag
     6721 ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata
     6781 aaaatatatc atcatgcctc ctctagacca gccaggacag aaatgcctcg acttcgctgc
     6841 tacccaaggt tgccgggtga cgcacaccgt ggaaacggat gaaggcacga acccagtgga
     6901 cataagcctg ttcggttcgt aagctgtaat gcaagtagcg tatgcgctca cgcaactggt
     6961 ccagaacctt gaccgaacgc agcggtggta acggcgcagt ggcggttttc atggcttgtt
     7021 atgactgttt ttttggggta cagtctatgc ctcgggcatc caagcagcaa gcgcgttacg
     7081 ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt tacgcagcag ggcagtcgcc
     7141 ctaaaacaaa gttaaacatt atgagggaag cggtgatcgc cgaagtatcg actcaactat
     7201 cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc gtacatttgt
     7261 acggctccgc agtggatggc ggcctgaagc cacacagtga tattgatttg ctggttacgg
     7321 tgaccgtaag gcttgatgaa acaacgcggc gagctttgat caacgacctt ttggaaactt
     7381 cggcttcccc tggagagagc gagattctcc gcgctgtaga agtcaccatt gttgtgcacg
     7441 acgacatcat tccgtggcgt tatccagcta agcgcgaact gcaatttgga gaatggcagc
     7501 gcaatgacat tcttgcaggt atcttcgagc cagccacgat cgacattgat ctggctatct
     7561 tgctgacaaa agcaagagaa catagcgttg ccttggtagg tccagcggcg gaggaactct
     7621 ttgatccggt tcctgaacag gatctatttg aggcgctaaa tgaaacctta acgctatgga
     7681 actcgccgcc cgactgggct ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt
     7741 ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg
     7801 agcgcctgcc ggcccagtat cagcccgtca tacttgaagc tagacaggct tatcttggac
     7861 aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc cactacgtga
     7921 aaggcgagat caccaaggta gtcggcaaat aaccctcgag ccacccatga ccaaaatccc
     7981 ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtag
//
