LOCUS       sequence-168595-        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.T237M
FEATURES             Location/Qualifiers
     source          1..8025
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      31..619
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     520..539
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     773..790
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(949..976)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1068..1154)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1203..1225
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1217..1234
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1218..1234
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1250..1349
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(6585..6684)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(6701..6720)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6702..6720)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6725..6741)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6725..6741)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             7172..7963
                     /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 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
       61 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
      121 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
      181 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
      241 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
      301 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
      361 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
      421 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
      481 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
      541 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
      601 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
      661 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
      721 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
      781 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
      841 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
      901 cgcaattaat acgcgtaccg ctagccagga agagtttgta gaaacgcaaa aaggccatcc
      961 gtcaggatgg ccttctgctt agtttgatgc ctggcagttt atggcgggcg tcctgcccgc
     1021 caccctccgg gccgttgctt cacaacgttc aaatccgctc ccggcggatt tgtcctactc
     1081 aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttc cgactgagcc
     1141 tttcgtttta tttgatgcct ggcagttccc tactctcgcg ttaacgctag catggatgtt
     1201 ttcccagtca cgacgttgta aaacgacggc cagtcttaag ctcgggcccc aaataatgat
     1261 tttattttga ctgatagtga cctgttcgtt gcaacaaatt gatgagcaat gcttttttat
     1321 aatgccaact ttgtacaaaa aagttggcac catggccact gattcagggg atccagccag
     1381 cacagaagat tctgagaaac ctgatggaat ttcatttgaa aacagagttc cccaggtcgc
     1441 tgcaactttg acagtagaag ctagactaaa ggagaaaaac agtaccttct ctgcttctgg
     1501 ggaaactgta gaaaggaaga gatttttccg aaagagtgtt gaaatgacgg aagatgacaa
     1561 agttgccgaa tcatccccca aagatgagag aattaaggct gcaatgaata ttccaagagt
     1621 agataagctt ccttcaaatg tgttgagagg tggacaagaa gttaaatatg aacagtgttc
     1681 aaagtcaacc tcagaaatct caaaagattg tttcaaggag aaaaatgaaa aggaaatgga
     1741 agaagaagca gaaatgaagg ctgtagctac ttctcctagt ggcagattcc tgaaatttga
     1801 catagaacta ggaagaggag catttaaaac agtatataaa ggactggaca ctgaaacatg
     1861 ggttgaggtt gcttggtgtg agctgcagga ccgaaagtta accaaagctg agcagcaaag
     1921 attcaaggaa gaagcagaga tgttgaaggg tctccagcac cccaatatag ttcgatttta
     1981 tgattcctgg gaatctatat taaaaggaaa gaaatgtatt gtattagtga ctgaactaat
     2041 gacatctggg accttaaaga tgtacttaaa acgatttaaa gtcatgaaac caaaggtctt
     2101 aaggagctgg tgcaggcaaa ttttaaaggg gttgcagttc ttgcacacta ggactcctcc
     2161 tattattcac cgggatctga agtgtgacaa tattttcatc acgggaccca ctggatctgt
     2221 gaagattggt gatctaggat tagccacctt aatgcgtacc tcatttgcta agagtgtcat
     2281 tggaactcct gagtttatgg ctccagagat gtatgaagaa cactatgatg aatccgtaga
     2341 tgtttatgct tttggaatgt gtatgctgga aatggccaca tcggagtatc cttattctga
     2401 gtgtcagaat gcagctcaaa tataccggaa agtaactagt ggcataaaac cagccagctt
     2461 caataaagtc actgatcctg aagtcaaaga aatcattgaa ggatgtattc gtcaaaacaa
     2521 atctgaaagg ttgtctatca gggacctatt aaaccatgca ttttttgctg aggatacagg
     2581 actgagggtg gagttagcag aagaagatga ttgctcaaat tcatcccttg ctttaagact
     2641 ctgggttgaa gaccctaaaa aattgaaagg caaacacaaa gacaatgaag ctattgaatt
     2701 tagtttcaac ttagaaacag atacacctga ggaagtagca tatgaaatgg tcaagtctgg
     2761 gttcttccat gaaagtgatt ccaaagctgt tgctaaatcc attagagacc gggtgacgcc
     2821 aataaagaag acaagagaga agaagcctgc tggctgtttg gaagaacgca gggattctca
     2881 gtgcaagtct atggggaatg tattccctca gccccagaat acaactttac cccttgctcc
     2941 cgctcagcaa actggggctg aatgtgaaga aactgaagtt gatcaacatg ttagacaaca
     3001 gcttctacaa agaaaaccac agcagcactg ctcctctgtt acaggtgaca atttgtctga
     3061 ggcaggagct gcatcagtta tacattcaga tacttcaagt cagcccagtg tagcctattc
     3121 ctcaaatcaa acgatgggct ctcaaatggt ttctaatatc ccgcaggctg aagtaaatgt
     3181 tccagggcaa atttattcat ctcagcaact agtaggacat taccagcaag tttcagggtt
     3241 acagaagcat tcaaagctga ctcagccgca gattttgcct ttggttcaag gtcagtccac
     3301 tgttttacct gtacatgtcc ttggaccgac agttgtttca caaccccagg tttccccatt
     3361 aactgttcag aaggtcccac agataaagcc tgtatcccaa ccagttggag ctgaacaaca
     3421 agcagctctt ctaaaaccag atttagttcg aagcttgaat caagatgtgg caactacgaa
     3481 ggaaaacgtc agtagccctg ataacccaag tggaaatggc aaacaggatc ggatcaaaca
     3541 gagaagagct tcctgtcccc gaccagagaa ggggactaaa tttcagctta ccgtccttca
     3601 ggtatcaacc tctggagata acatggtaga gtgtcagctg gagacacaca acaacaagat
     3661 ggtcaccttc aagtttgatg ttgatggtga tgcgccagag gatattgcag actatatggt
     3721 tgaagataac tttgtgctgg aaagtgagaa agaaaaattt gtagaagaat tgagagctat
     3781 tgtaggtcaa gcccaggaga tccttcatgt ccactttgcc acagaaagag ccactggagt
     3841 tgattctatt actgtggact ccaacagtag ccagacaggg tcatctgaac aagtacagat
     3901 aaattctaca tctactcaaa ccagcaatga atctgctcct cagtcatccc cagttggtcg
     3961 gtggcgattc tgtatcaatc agacgataag aaaccgtgag actcagtctc ctccttctct
     4021 tcagcattcc atgtctgcgg ttcctggccg acatccactt cctagtccaa aaaacacaag
     4081 taataaggaa atatcacggg acacattgct cactatagaa aataatccat gccaccgtgc
     4141 acttttcacc tccaaatcag aacacaagga tgtggttgat ggtaagattt ctgaatgtgc
     4201 tagtgtagaa accaagcagc cagctatact ttatcaagtg gaagataaca ggcagataat
     4261 ggcaccagtt actaatagtt ccagttactc tactacttca gttcgtgcag ttccagctga
     4321 atgtgaggga ctcaccaaac aagcaagcat attcatacct gtgtatccat gtcaccaaac
     4381 tgccagtcag gctgatgcac ttatgtccca tcctggcgaa tcaactcaga cttctggtaa
     4441 ctctcttaca actctggcat ttgatcaaaa gcctcaaacc ttatcagtac agcagccagc
     4501 tatggatgca gagtttattt ctcaagaagg agaaactaca gtgaacactg aagcaagttc
     4561 tcctaagaca gtcattccca ctcagacccc tggccttgaa ccaactaccc ttcaacccac
     4621 tactgtcctg gaatcagatg gagaaagacc tccaaaactg gagtttgcag acaaccgaat
     4681 taaaactctg gatgaaaaat taagaaactt gctctatcag gagcacagca tctctagcat
     4741 ctatcccgag agtcagaagg atacccaaag catagactct ccattttctt cctctgctga
     4801 agataccctc tcctgtccag tgacagaagt catagccatc agtcactgtg gaattaaaga
     4861 tagccctgta caatccccta atttccaaca gacaggctct aagcttctgt ccaatgtggc
     4921 tgcaagtcag cctgctaata tatcagtgtt caaaagggac ctgaatgtga taacttctgt
     4981 acccagcgaa ttgtgtttac atgagatgtc ctcagatgct tcacttccag gggatccaga
     5041 ggcctatcct gctgctgtgt caagcggtgg agccattcat ctgcagacag gagtggaaac
     5101 agaagagatg agatcagcaa ttgctcctga tcccattcct ctgacacggg agtccacagc
     5161 tgatactagg gctttgaata gatgtaaagc gatgagtgga tcatttcagc ggggtcggtt
     5221 ccaggtgatt acaattcctc agcagcagtc agcaaaaatg acatcttttg gaatagaaca
     5281 catatcagtg ttcagtgaga caaaccattc tagtgaagaa gcctttatta aaacagcaaa
     5341 gtctcagttg gtagaaatag aacctgccac acaaaatcca aaaacttcgt tttcttatga
     5401 gaagttacaa gctcttcagg aaacctgtaa agaaaataaa ggagttccca aacaaggtga
     5461 caacttctta tctttcagcg cagcttgtga gactgatgta tcttcagtga ccccagaaaa
     5521 ggaatttgaa gaaacttcag ccacaggaag tagcatgcag tctggatctg aactgttgct
     5581 taaagagaga gagatattga ctgctgggaa acagcctagc tctgatagtg aattttcagc
     5641 cagtcttgct ggcagtggaa agtcagtggc aaagactggt ccagagagta atcagtgctt
     5701 accacaccac gaagaacaag cttatgctca aacacagagt tcactcttct attcgccatc
     5761 ttccccaatg agcagtgatg atgaatcaga aatagaggat gaggacttga aggtggagct
     5821 tcaaagatta cgagaaaaac acattcagga ggtggtaaat cttcaaaccc agcagaataa
     5881 ggagctgcag gagctctatg aacgccttcg gtcaattaaa gatagcaaaa cccaatctac
     5941 tgagattcct ttgccacctg catcaccacg tcgaccaaga tctttcaaaa gcaaacttcg
     6001 aagccgcccc cagtccttga cacatgtgga caatggcata gttgctacag atccactgtg
     6061 tgtggagagt aatgcagcat catgccaaca gtctccagcc agtaaaaaag ggatgttcac
     6121 agatgactta cacaagctgg tggatgactg gacaaaggaa gcagtaggaa attctcttat
     6181 taagccaagt ttaaaccaac ttaaacaaag tcaacacaaa ctagagacag aaaactggaa
     6241 taaagtatct gaaaatactc cgtctactat gggctacaca tcaacatgga tttcttctct
     6301 gtcccaaatc cgtggagctg tcccaacttc cttgccacaa ggactctcac tcccttcatt
     6361 tcctgggcca ttatcatcat atggaatgcc tcacgtttgt cagtataatg ctgtggcggg
     6421 ggcggggtat ccagtacagt gggtaggaat ttcaggaaca acacaacaat ctgtagtaat
     6481 tcccgcccaa tctgggggac cattccagcc agggatgaat atgcaggcat ttccaacttc
     6541 atcagtgcag aatcctgcca caatccctcc tggtcctaaa tgattgccaa ctttcttgta
     6601 caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat
     6661 cagtcaaaat aaaatcatta tttgccatcc agctgatatc ccctatagtg agtcgtatta
     6721 catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat
     6781 tgcacaagat aaaaatatat catcatgcct cctctagacc agccaggaca gaaatgcctc
     6841 gacttcgctg ctacccaagg ttgccgggtg acgcacaccg tggaaacgga tgaaggcacg
     6901 aacccagtgg acataagcct gttcggttcg taagctgtaa tgcaagtagc gtatgcgctc
     6961 acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt
     7021 catggcttgt tatgactgtt tttttggggt acagtctatg cctcgggcat ccaagcagca
     7081 agcgcgttac gccgtgggtc gatgtttgat gttatggagc agcaacgatg ttacgcagca
     7141 gggcagtcgc cctaaaacaa agttaaacat tatgagggaa gcggtgatcg ccgaagtatc
     7201 gactcaacta tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc
     7261 cgtacatttg tacggctccg cagtggatgg cggcctgaag ccacacagtg atattgattt
     7321 gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga tcaacgacct
     7381 tttggaaact tcggcttccc ctggagagag cgagattctc cgcgctgtag aagtcaccat
     7441 tgttgtgcac gacgacatca ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg
     7501 agaatggcag cgcaatgaca ttcttgcagg tatcttcgag ccagccacga tcgacattga
     7561 tctggctatc ttgctgacaa aagcaagaga acatagcgtt gccttggtag gtccagcggc
     7621 ggaggaactc tttgatccgg ttcctgaaca ggatctattt gaggcgctaa atgaaacctt
     7681 aacgctatgg aactcgccgc ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt
     7741 gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga
     7801 ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc atacttgaag ctagacaggc
     7861 ttatcttgga caagaagaag atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt
     7921 ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa taaccctcga gccacccatg
     7981 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtc
//
