LOCUS       sequence-168662-        8025 bp    DNA     circular SYN 15-APR-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 Apr 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.S617F
FEATURES             Location/Qualifiers
     source          1..8025
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      69..657
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     558..577
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     811..828
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(987..1014)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1106..1192)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1241..1263
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1255..1272
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1256..1272
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1288..1387
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(6623..6722)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(6739..6758)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6740..6758)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6763..6779)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6763..6779)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             7210..8001
                     /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 tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa
       61 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
      121 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
      181 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
      241 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
      301 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
      361 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
      421 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
      481 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
      541 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
      601 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
      661 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
      721 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
      781 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
      841 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
      901 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct
      961 agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag
     1021 tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca
     1081 caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac
     1141 aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg
     1201 cagttcccta ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa
     1261 acgacggcca gtcttaagct cgggccccaa ataatgattt tattttgact gatagtgacc
     1321 tgttcgttgc aacaaattga tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa
     1381 gttggcacca tggccactga ttcaggggat ccagccagca cagaagattc tgagaaacct
     1441 gatggaattt catttgaaaa cagagttccc caggtcgctg caactttgac agtagaagct
     1501 agactaaagg agaaaaacag taccttctct gcttctgggg aaactgtaga aaggaagaga
     1561 tttttccgaa agagtgttga aatgacggaa gatgacaaag ttgccgaatc atcccccaaa
     1621 gatgagagaa ttaaggctgc aatgaatatt ccaagagtag ataagcttcc ttcaaatgtg
     1681 ttgagaggtg gacaagaagt taaatatgaa cagtgttcaa agtcaacctc agaaatctca
     1741 aaagattgtt tcaaggagaa aaatgaaaag gaaatggaag aagaagcaga aatgaaggct
     1801 gtagctactt ctcctagtgg cagattcctg aaatttgaca tagaactagg aagaggagca
     1861 tttaaaacag tatataaagg actggacact gaaacatggg ttgaggttgc ttggtgtgag
     1921 ctgcaggacc gaaagttaac caaagctgag cagcaaagat tcaaggaaga agcagagatg
     1981 ttgaagggtc tccagcaccc caatatagtt cgattttatg attcctggga atctatatta
     2041 aaaggaaaga aatgtattgt attagtgact gaactaatga catctgggac cttaaagacg
     2101 tacttaaaac gatttaaagt catgaaacca aaggtcttaa ggagctggtg caggcaaatt
     2161 ttaaaggggt tgcagttctt gcacactagg actcctccta ttattcaccg ggatctgaag
     2221 tgtgacaata ttttcatcac gggacccact ggatctgtga agattggtga tctaggatta
     2281 gccaccttaa tgcgtacctc atttgctaag agtgtcattg gaactcctga gtttatggct
     2341 ccagagatgt atgaagaaca ctatgatgaa tccgtagatg tttatgcttt tggaatgtgt
     2401 atgctggaaa tggccacatc ggagtatcct tattctgagt gtcagaatgc agctcaaata
     2461 taccggaaag taactagtgg cataaaacca gccagcttca ataaagtcac tgatcctgaa
     2521 gtcaaagaaa tcattgaagg atgtattcgt caaaacaaat ctgaaaggtt gtctatcagg
     2581 gacctattaa accatgcatt ttttgctgag gatacaggac tgagggtgga gttagcagaa
     2641 gaagatgatt gctcaaattc atcccttgct ttaagactct gggttgaaga ccctaaaaaa
     2701 ttgaaaggca aacacaaaga caatgaagct attgaattta gtttcaactt agaaacagat
     2761 acacctgagg aagtagcata tgaaatggtc aagtctgggt tcttccatga aagtgattcc
     2821 aaagctgttg ctaaatccat tagagaccgg gtgacgccaa taaagaagac aagagagaag
     2881 aagcctgctg gctgtttgga agaacgcagg gattctcagt gcaagtctat ggggaatgta
     2941 ttccctcagc cccagaatac aactttaccc cttgctcccg ctcagcaaac tggggctgaa
     3001 tgtgaagaaa ctgaagttga tcaacatgtt agacaacagc ttctacaaag aaaaccacag
     3061 cagcactgct cctctgttac aggtgacaat ttgtctgagg caggagctgc atcagttata
     3121 cattcagata cttcaagtca gcccagtgta gcctattcct caaatcaaac gatgggctct
     3181 caaatggttt ctaatatccc gcaggctgaa gtaaatgttc cagggcaaat ttattcattt
     3241 cagcaactag taggacatta ccagcaagtt tcagggttac agaagcattc aaagctgact
     3301 cagccgcaga ttttgccttt ggttcaaggt cagtccactg ttttacctgt acatgtcctt
     3361 ggaccgacag ttgtttcaca accccaggtt tccccattaa ctgttcagaa ggtcccacag
     3421 ataaagcctg tatcccaacc agttggagct gaacaacaag cagctcttct aaaaccagat
     3481 ttagttcgaa gcttgaatca agatgtggca actacgaagg aaaacgtcag tagccctgat
     3541 aacccaagtg gaaatggcaa acaggatcgg atcaaacaga gaagagcttc ctgtccccga
     3601 ccagagaagg ggactaaatt tcagcttacc gtccttcagg tatcaacctc tggagataac
     3661 atggtagagt gtcagctgga gacacacaac aacaagatgg tcaccttcaa gtttgatgtt
     3721 gatggtgatg cgccagagga tattgcagac tatatggttg aagataactt tgtgctggaa
     3781 agtgagaaag aaaaatttgt agaagaattg agagctattg taggtcaagc ccaggagatc
     3841 cttcatgtcc actttgccac agaaagagcc actggagttg attctattac tgtggactcc
     3901 aacagtagcc agacagggtc atctgaacaa gtacagataa attctacatc tactcaaacc
     3961 agcaatgaat ctgctcctca gtcatcccca gttggtcggt ggcgattctg tatcaatcag
     4021 acgataagaa accgtgagac tcagtctcct ccttctcttc agcattccat gtctgcggtt
     4081 cctggccgac atccacttcc tagtccaaaa aacacaagta ataaggaaat atcacgggac
     4141 acattgctca ctatagaaaa taatccatgc caccgtgcac ttttcacctc caaatcagaa
     4201 cacaaggatg tggttgatgg taagatttct gaatgtgcta gtgtagaaac caagcagcca
     4261 gctatacttt atcaagtgga agataacagg cagataatgg caccagttac taatagttcc
     4321 agttactcta ctacttcagt tcgtgcagtt ccagctgaat gtgagggact caccaaacaa
     4381 gcaagcatat tcatacctgt gtatccatgt caccaaactg ccagtcaggc tgatgcactt
     4441 atgtcccatc ctggcgaatc aactcagact tctggtaact ctcttacaac tctggcattt
     4501 gatcaaaagc ctcaaacctt atcagtacag cagccagcta tggatgcaga gtttatttct
     4561 caagaaggag aaactacagt gaacactgaa gcaagttctc ctaagacagt cattcccact
     4621 cagacccctg gccttgaacc aactaccctt caacccacta ctgtcctgga atcagatgga
     4681 gaaagacctc caaaactgga gtttgcagac aaccgaatta aaactctgga tgaaaaatta
     4741 agaaacttgc tctatcagga gcacagcatc tctagcatct atcccgagag tcagaaggat
     4801 acccaaagca tagactctcc attttcttcc tctgctgaag ataccctctc ctgtccagtg
     4861 acagaagtca tagccatcag tcactgtgga attaaagata gccctgtaca atcccctaat
     4921 ttccaacaga caggctctaa gcttctgtcc aatgtggctg caagtcagcc tgctaatata
     4981 tcagtgttca aaagggacct gaatgtgata acttctgtac ccagcgaatt gtgtttacat
     5041 gagatgtcct cagatgcttc acttccaggg gatccagagg cctatcctgc tgctgtgtca
     5101 agcggtggag ccattcatct gcagacagga gtggaaacag aagagatgag atcagcaatt
     5161 gctcctgatc ccattcctct gacacgggag tccacagctg atactagggc tttgaataga
     5221 tgtaaagcga tgagtggatc atttcagcgg ggtcggttcc aggtgattac aattcctcag
     5281 cagcagtcag caaaaatgac atcttttgga atagaacaca tatcagtgtt cagtgagaca
     5341 aaccattcta gtgaagaagc ctttattaaa acagcaaagt ctcagttggt agaaatagaa
     5401 cctgccacac aaaatccaaa aacttcgttt tcttatgaga agttacaagc tcttcaggaa
     5461 acctgtaaag aaaataaagg agttcccaaa caaggtgaca acttcttatc tttcagcgca
     5521 gcttgtgaga ctgatgtatc ttcagtgacc ccagaaaagg aatttgaaga aacttcagcc
     5581 acaggaagta gcatgcagtc tggatctgaa ctgttgctta aagagagaga gatattgact
     5641 gctgggaaac agcctagctc tgatagtgaa ttttcagcca gtcttgctgg cagtggaaag
     5701 tcagtggcaa agactggtcc agagagtaat cagtgcttac cacaccacga agaacaagct
     5761 tatgctcaaa cacagagttc actcttctat tcgccatctt ccccaatgag cagtgatgat
     5821 gaatcagaaa tagaggatga ggacttgaag gtggagcttc aaagattacg agaaaaacac
     5881 attcaggagg tggtaaatct tcaaacccag cagaataagg agctgcagga gctctatgaa
     5941 cgccttcggt caattaaaga tagcaaaacc caatctactg agattccttt gccacctgca
     6001 tcaccacgtc gaccaagatc tttcaaaagc aaacttcgaa gccgccccca gtccttgaca
     6061 catgtggaca atggcatagt tgctacagat ccactgtgtg tggagagtaa tgcagcatca
     6121 tgccaacagt ctccagccag taaaaaaggg atgttcacag atgacttaca caagctggtg
     6181 gatgactgga caaaggaagc agtaggaaat tctcttatta agccaagttt aaaccaactt
     6241 aaacaaagtc aacacaaact agagacagaa aactggaata aagtatctga aaatactccg
     6301 tctactatgg gctacacatc aacatggatt tcttctctgt cccaaatccg tggagctgtc
     6361 ccaacttcct tgccacaagg actctcactc ccttcatttc ctgggccatt atcatcatat
     6421 ggaatgcctc acgtttgtca gtataatgct gtggcggggg cggggtatcc agtacagtgg
     6481 gtaggaattt caggaacaac acaacaatct gtagtaattc ccgcccaatc tgggggacca
     6541 ttccagccag ggatgaatat gcaggcattt ccaacttcat cagtgcagaa tcctgccaca
     6601 atccctcctg gtcctaaatg attgccaact ttcttgtaca aagttggcat tataagaaag
     6661 cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt
     6721 tgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg
     6781 cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca
     6841 tcatgcctcc tctagaccag ccaggacaga aatgcctcga cttcgctgct acccaaggtt
     6901 gccgggtgac gcacaccgtg gaaacggatg aaggcacgaa cccagtggac ataagcctgt
     6961 tcggttcgta agctgtaatg caagtagcgt atgcgctcac gcaactggtc cagaaccttg
     7021 accgaacgca gcggtggtaa cggcgcagtg gcggttttca tggcttgtta tgactgtttt
     7081 tttggggtac agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga
     7141 tgtttgatgt tatggagcag caacgatgtt acgcagcagg gcagtcgccc taaaacaaag
     7201 ttaaacatta tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt
     7261 ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg tacatttgta cggctccgca
     7321 gtggatggcg gcctgaagcc acacagtgat attgatttgc tggttacggt gaccgtaagg
     7381 cttgatgaaa caacgcggcg agctttgatc aacgaccttt tggaaacttc ggcttcccct
     7441 ggagagagcg agattctccg cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt
     7501 ccgtggcgtt atccagctaa gcgcgaactg caatttggag aatggcagcg caatgacatt
     7561 cttgcaggta tcttcgagcc agccacgatc gacattgatc tggctatctt gctgacaaaa
     7621 gcaagagaac atagcgttgc cttggtaggt ccagcggcgg aggaactctt tgatccggtt
     7681 cctgaacagg atctatttga ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc
     7741 gactgggctg gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg gtacagcgca
     7801 gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga gcgcctgccg
     7861 gcccagtatc agcccgtcat acttgaagct agacaggctt atcttggaca agaagaagat
     7921 cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc actacgtgaa aggcgagatc
     7981 accaaggtag tcggcaaata accctcgagc cacccatgac caaaa
//
