LOCUS       sequence-168753-        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.V488A
FEATURES             Location/Qualifiers
     source          1..8025
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      42..630
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     531..550
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     784..801
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(960..987)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1079..1165)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1214..1236
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1228..1245
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1229..1245
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1261..1360
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(6596..6695)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(6712..6731)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6713..6731)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6736..6752)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6736..6752)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             7183..7974
                     /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 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
       61 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
      121 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
      181 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
      241 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
      301 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
      361 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
      421 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
      481 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
      541 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
      601 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
      661 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
      721 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
      781 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
      841 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
      901 agtgagcgca acgcaattaa tacgcgtacc gctagccagg aagagtttgt agaaacgcaa
      961 aaaggccatc cgtcaggatg gccttctgct tagtttgatg cctggcagtt tatggcgggc
     1021 gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat
     1081 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
     1141 ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta
     1201 gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc
     1261 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
     1321 tgctttttta taatgccaac tttgtacaaa aaagttggca ccatggccac tgattcaggg
     1381 gatccagcca gcacagaaga ttctgagaaa cctgatggaa tttcatttga aaacagagtt
     1441 ccccaggtcg ctgcaacttt gacagtagaa gctagactaa aggagaaaaa cagtaccttc
     1501 tctgcttctg gggaaactgt agaaaggaag agatttttcc gaaagagtgt tgaaatgacg
     1561 gaagatgaca aagttgccga atcatccccc aaagatgaga gaattaaggc tgcaatgaat
     1621 attccaagag tagataagct tccttcaaat gtgttgagag gtggacaaga agttaaatat
     1681 gaacagtgtt caaagtcaac ctcagaaatc tcaaaagatt gtttcaagga gaaaaatgaa
     1741 aaggaaatgg aagaagaagc agaaatgaag gctgtagcta cttctcctag tggcagattc
     1801 ctgaaatttg acatagaact aggaagagga gcatttaaaa cagtatataa aggactggac
     1861 actgaaacat gggttgaggt tgcttggtgt gagctgcagg accgaaagtt aaccaaagct
     1921 gagcagcaaa gattcaagga agaagcagag atgttgaagg gtctccagca ccccaatata
     1981 gttcgatttt atgattcctg ggaatctata ttaaaaggaa agaaatgtat tgtattagtg
     2041 actgaactaa tgacatctgg gaccttaaag acgtacttaa aacgatttaa agtcatgaaa
     2101 ccaaaggtct taaggagctg gtgcaggcaa attttaaagg ggttgcagtt cttgcacact
     2161 aggactcctc ctattattca ccgggatctg aagtgtgaca atattttcat cacgggaccc
     2221 actggatctg tgaagattgg tgatctagga ttagccacct taatgcgtac ctcatttgct
     2281 aagagtgtca ttggaactcc tgagtttatg gctccagaga tgtatgaaga acactatgat
     2341 gaatccgtag atgtttatgc ttttggaatg tgtatgctgg aaatggccac atcggagtat
     2401 ccttattctg agtgtcagaa tgcagctcaa atataccgga aagtaactag tggcataaaa
     2461 ccagccagct tcaataaagt cactgatcct gaagtcaaag aaatcattga aggatgtatt
     2521 cgtcaaaaca aatctgaaag gttgtctatc agggacctat taaaccatgc attttttgct
     2581 gaggatacag gactgagggt ggagttagca gaagaagatg attgctcaaa ttcatccctt
     2641 gctttaagac tctgggttga agaccctaaa aaattgaaag gcaaacacaa agacaatgaa
     2701 gctattgaat ttagtttcaa cttagaaaca gatacacctg aggaagtagc atatgaaatg
     2761 gtcaagtctg ggttcttcca tgaaagtgat tccaaagctg ttgctaaatc cattagagac
     2821 cgggcgacgc caataaagaa gacaagagag aagaagcctg ctggctgttt ggaagaacgc
     2881 agggattctc agtgcaagtc tatggggaat gtattccctc agccccagaa tacaacttta
     2941 ccccttgctc ccgctcagca aactggggct gaatgtgaag aaactgaagt tgatcaacat
     3001 gttagacaac agcttctaca aagaaaacca cagcagcact gctcctctgt tacaggtgac
     3061 aatttgtctg aggcaggagc tgcatcagtt atacattcag atacttcaag tcagcccagt
     3121 gtagcctatt cctcaaatca aacgatgggc tctcaaatgg tttctaatat cccgcaggct
     3181 gaagtaaatg ttccagggca aatttattca tctcagcaac tagtaggaca ttaccagcaa
     3241 gtttcagggt tacagaagca ttcaaagctg actcagccgc agattttgcc tttggttcaa
     3301 ggtcagtcca ctgttttacc tgtacatgtc cttggaccga cagttgtttc acaaccccag
     3361 gtttccccat taactgttca gaaggtccca cagataaagc ctgtatccca accagttgga
     3421 gctgaacaac aagcagctct tctaaaacca gatttagttc gaagcttgaa tcaagatgtg
     3481 gcaactacga aggaaaacgt cagtagccct gataacccaa gtggaaatgg caaacaggat
     3541 cggatcaaac agagaagagc ttcctgtccc cgaccagaga aggggactaa atttcagctt
     3601 accgtccttc aggtatcaac ctctggagat aacatggtag agtgtcagct ggagacacac
     3661 aacaacaaga tggtcacctt caagtttgat gttgatggtg atgcgccaga ggatattgca
     3721 gactatatgg ttgaagataa ctttgtgctg gaaagtgaga aagaaaaatt tgtagaagaa
     3781 ttgagagcta ttgtaggtca agcccaggag atccttcatg tccactttgc cacagaaaga
     3841 gccactggag ttgattctat tactgtggac tccaacagta gccagacagg gtcatctgaa
     3901 caagtacaga taaattctac atctactcaa accagcaatg aatctgctcc tcagtcatcc
     3961 ccagttggtc ggtggcgatt ctgtatcaat cagacgataa gaaaccgtga gactcagtct
     4021 cctccttctc ttcagcattc catgtctgcg gttcctggcc gacatccact tcctagtcca
     4081 aaaaacacaa gtaataagga aatatcacgg gacacattgc tcactataga aaataatcca
     4141 tgccaccgtg cacttttcac ctccaaatca gaacacaagg atgtggttga tggtaagatt
     4201 tctgaatgtg ctagtgtaga aaccaagcag ccagctatac tttatcaagt ggaagataac
     4261 aggcagataa tggcaccagt tactaatagt tccagttact ctactacttc agttcgtgca
     4321 gttccagctg aatgtgaggg actcaccaaa caagcaagca tattcatacc tgtgtatcca
     4381 tgtcaccaaa ctgccagtca ggctgatgca cttatgtccc atcctggcga atcaactcag
     4441 acttctggta actctcttac aactctggca tttgatcaaa agcctcaaac cttatcagta
     4501 cagcagccag ctatggatgc agagtttatt tctcaagaag gagaaactac agtgaacact
     4561 gaagcaagtt ctcctaagac agtcattccc actcagaccc ctggccttga accaactacc
     4621 cttcaaccca ctactgtcct ggaatcagat ggagaaagac ctccaaaact ggagtttgca
     4681 gacaaccgaa ttaaaactct ggatgaaaaa ttaagaaact tgctctatca ggagcacagc
     4741 atctctagca tctatcccga gagtcagaag gatacccaaa gcatagactc tccattttct
     4801 tcctctgctg aagataccct ctcctgtcca gtgacagaag tcatagccat cagtcactgt
     4861 ggaattaaag atagccctgt acaatcccct aatttccaac agacaggctc taagcttctg
     4921 tccaatgtgg ctgcaagtca gcctgctaat atatcagtgt tcaaaaggga cctgaatgtg
     4981 ataacttctg tacccagcga attgtgttta catgagatgt cctcagatgc ttcacttcca
     5041 ggggatccag aggcctatcc tgctgctgtg tcaagcggtg gagccattca tctgcagaca
     5101 ggagtggaaa cagaagagat gagatcagca attgctcctg atcccattcc tctgacacgg
     5161 gagtccacag ctgatactag ggctttgaat agatgtaaag cgatgagtgg atcatttcag
     5221 cggggtcggt tccaggtgat tacaattcct cagcagcagt cagcaaaaat gacatctttt
     5281 ggaatagaac acatatcagt gttcagtgag acaaaccatt ctagtgaaga agcctttatt
     5341 aaaacagcaa agtctcagtt ggtagaaata gaacctgcca cacaaaatcc aaaaacttcg
     5401 ttttcttatg agaagttaca agctcttcag gaaacctgta aagaaaataa aggagttccc
     5461 aaacaaggtg acaacttctt atctttcagc gcagcttgtg agactgatgt atcttcagtg
     5521 accccagaaa aggaatttga agaaacttca gccacaggaa gtagcatgca gtctggatct
     5581 gaactgttgc ttaaagagag agagatattg actgctggga aacagcctag ctctgatagt
     5641 gaattttcag ccagtcttgc tggcagtgga aagtcagtgg caaagactgg tccagagagt
     5701 aatcagtgct taccacacca cgaagaacaa gcttatgctc aaacacagag ttcactcttc
     5761 tattcgccat cttccccaat gagcagtgat gatgaatcag aaatagagga tgaggacttg
     5821 aaggtggagc ttcaaagatt acgagaaaaa cacattcagg aggtggtaaa tcttcaaacc
     5881 cagcagaata aggagctgca ggagctctat gaacgccttc ggtcaattaa agatagcaaa
     5941 acccaatcta ctgagattcc tttgccacct gcatcaccac gtcgaccaag atctttcaaa
     6001 agcaaacttc gaagccgccc ccagtccttg acacatgtgg acaatggcat agttgctaca
     6061 gatccactgt gtgtggagag taatgcagca tcatgccaac agtctccagc cagtaaaaaa
     6121 gggatgttca cagatgactt acacaagctg gtggatgact ggacaaagga agcagtagga
     6181 aattctctta ttaagccaag tttaaaccaa cttaaacaaa gtcaacacaa actagagaca
     6241 gaaaactgga ataaagtatc tgaaaatact ccgtctacta tgggctacac atcaacatgg
     6301 atttcttctc tgtcccaaat ccgtggagct gtcccaactt ccttgccaca aggactctca
     6361 ctcccttcat ttcctgggcc attatcatca tatggaatgc ctcacgtttg tcagtataat
     6421 gctgtggcgg gggcggggta tccagtacag tgggtaggaa tttcaggaac aacacaacaa
     6481 tctgtagtaa ttcccgccca atctggggga ccattccagc cagggatgaa tatgcaggca
     6541 tttccaactt catcagtgca gaatcctgcc acaatccctc ctggtcctaa atgattgcca
     6601 actttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa
     6661 caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt
     6721 gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc
     6781 tgatgttaca ttgcacaaga taaaaatata tcatcatgcc tcctctagac cagccaggac
     6841 agaaatgcct cgacttcgct gctacccaag gttgccgggt gacgcacacc gtggaaacgg
     6901 atgaaggcac gaacccagtg gacataagcc tgttcggttc gtaagctgta atgcaagtag
     6961 cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca
     7021 gtggcggttt tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca
     7081 tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat
     7141 gttacgcagc agggcagtcg ccctaaaaca aagttaaaca ttatgaggga agcggtgatc
     7201 gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg
     7261 acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt
     7321 gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg
     7381 atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta
     7441 gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa
     7501 ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg
     7561 atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta
     7621 ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta
     7681 aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta
     7741 gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat
     7801 gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa
     7861 gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg
     7921 gaagaatttg tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg
     7981 agccacccat gaccaaaatc ccttaacgtg agttacgcgt cgttc
//
