LOCUS       sequence-165308-        6960 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing MET , part of the Target Accelerator
            Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6960)
  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 6960)
  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_MET_WT
FEATURES             Location/Qualifiers
     source          1..6960
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     43..65
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     57..74
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     58..74
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    90..189
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(4360..4459)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4476..4495)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4477..4495)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4500..4516)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4500..4516)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4947..5738
                     /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      5831..6419
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6320..6339
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6573..6590
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(6749..6776)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6868..6954)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta
       61 aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga
      121 cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa
      181 aagttggcac catgaaggcc cccgctgtgc ttgcacctgg catcctcgtg ctcctgttta
      241 ccttggtgca gaggagcaat ggggagtgta aagaggcact agcaaagtcc gagatgaatg
      301 tgaatatgaa gtatcagctt cccaacttca ccgcggaaac acccatccag aatgtcattc
      361 tacatgagca tcacattttc cttggtgcca ctaactacat ttatgtttta aatgaggaag
      421 accttcagaa ggttgctgag tacaagactg ggcctgtgct ggaacaccca gattgtttcc
      481 catgtcagga ctgcagcagc aaagccaatt tatcaggagg tgtttggaaa gataacatca
      541 acatggctct agttgtcgac acctactatg atgatcaact cattagctgt ggcagcgtca
      601 acagagggac ctgccagcga catgtctttc cccacaatca tactgctgac atacagtcgg
      661 aggttcactg catattctcc ccacagatag aagagcccag ccagtgtcct gactgtgtgg
      721 tgagcgccct gggagccaaa gtcctttcat ctgtaaagga ccggttcatc aacttctttg
      781 taggcaatac cataaattct tcttatttcc cagatcatcc attgcattcg atatcagtga
      841 gaaggctaaa ggaaacgaaa gatggtttta tgtttttgac ggaccagtcc tacattgatg
      901 ttttacctga gttcagagat tcttacccca ttaagtatgt ccatgccttt gaaagcaaca
      961 attttattta cttcttgacg gtccaaaggg aaactctaga tgctcagact tttcacacaa
     1021 gaataatcag gttctgttcc ataaactctg gattgcattc ctacatggaa atgcctctgg
     1081 agtgtattct cacagaaaag agaaaaaaga gatccacaaa gaaggaagtg tttaatatac
     1141 ttcaggctgc gtatgtcagc aagcctgggg cccagcttgc tagacaaata ggagccagcc
     1201 tgaatgatga cattcttttc ggggtgttcg cacaaagcaa gccagattct gccgaaccaa
     1261 tggatcgatc tgccatgtgt gcattcccta tcaaatatgt caacgacttc ttcaacaaga
     1321 tcgtcaacaa aaacaatgtg agatgtctcc agcattttta cggacccaat catgagcact
     1381 gctttaatag gacacttctg agaaattcat caggctgtga agcgcgccgt gatgaatatc
     1441 gaacagagtt taccacagct ttgcagcgcg ttgacttatt catgggtcaa ttcagcgaag
     1501 tcctcttaac atctatatcc accttcatta aaggagacct caccatagct aatcttggga
     1561 catcagaggg tcgcttcatg caggttgtgg tttctcgatc aggaccatca acccctcatg
     1621 tgaattttct cctggactcc catccagtgt ctccagaagt gattgtggag catacattaa
     1681 accaaaatgg ctacacactg gttatcactg ggaagaagat cacgaagatc ccattgaatg
     1741 gcttgggctg cagacatttc cagtcctgca gtcaatgcct ctctgcccca ccctttgttc
     1801 agtgtggctg gtgccacgac aaatgtgtgc gatcggagga atgcctgagc gggacatgga
     1861 ctcaacagat ctgtctgcct gcaatctaca aggttttccc aaatagtgca ccccttgaag
     1921 gagggacaag gctgaccata tgtggctggg actttggatt tcggaggaat aataaatttg
     1981 atttaaagaa aactagagtt ctccttggaa atgagagctg caccttgact ttaagtgaga
     2041 gcacgatgaa tacattgaaa tgcacagttg gtcctgccat gaataagcat ttcaatatgt
     2101 ccataattat ttcaaatggc cacgggacaa cacagtacag tacattctcc tatgtggatc
     2161 ctgtaataac aagtatttcg ccgaaatacg gtcctatggc tggtggcact ttacttactt
     2221 taactggaaa ttacctaaac agtgggaatt ctagacacat ttcaattggt ggaaaaacat
     2281 gtactttaaa aagtgtgtca aacaatattc ttgaatgtta taccccagcc caaaccattt
     2341 caactgagtt tgctgttaaa ttgaaaattg acttagccaa ccgagagaca agcatcttca
     2401 gttaccgtga agatcccatt gtctatgaaa ttcatccaac caaatctttt attagtggtg
     2461 ggagcacaat aacaggtgtt gggaaaaacc tgaattcagt tagtgtcccg agaatggtca
     2521 taaatgtgca tgaagcagga aggaacttta cagtggcatg tcaacatcgc tctaattcag
     2581 agataatctg ttgtaccact ccttccctgc aacagctgaa tctgcaactc cccctgaaaa
     2641 ccaaagcctt tttcatgtta gatgggatcc tttccaaata ctttgatctc atttatgtac
     2701 ataatcctgt gtttaagcct tttgaaaagc cagtgatgat ctcaatgggc aatgaaaatg
     2761 tactggaaat taagggaaat gatattgacc ctgaagcagt taaaggtgaa gtgttaaaag
     2821 ttggaaataa gagctgtgag aatatacact tacattctga agccgtttta tgcacggtcc
     2881 ccaatgacct gctgaaattg aacagcgagc taaatataga gtggaagcaa gcaatttctt
     2941 caaccgtcct tggaaaagta atagttcaac cagatcagaa tttcacagga ttgattgctg
     3001 gtgttgtctc aatatcaaca gcactgttat tactacttgg gtttttcctg tggctgaaaa
     3061 agagaaagca aattaaagat ctgggcagtg aattagttcg ctacgatgca agagtacaca
     3121 ctcctcattt ggataggctt gtaagtgccc gaagtgtaag cccaactaca gaaatggttt
     3181 caaatgaatc tgtagactac cgagctactt ttccagaaga tcagtttcct aattcatctc
     3241 agaacggttc atgccgacaa gtgcagtatc ctctgacaga catgtccccc atcctaacta
     3301 gtggggactc tgatatatcc agtccattac tgcaaaatac tgtccacatt gacctcagtg
     3361 ctctaaatcc agagctggtc caggcagtgc agcatgtagt gattgggccc agtagcctga
     3421 ttgtgcattt caatgaagtc ataggaagag ggcattttgg ttgtgtatat catgggactt
     3481 tgttggacaa tgatggcaag aaaattcact gtgctgtgaa atccttgaac agaatcactg
     3541 acataggaga agtttcccaa tttctgaccg agggaatcat catgaaagat tttagtcatc
     3601 ccaatgtcct ctcgctcctg ggaatctgcc tgcgaagtga agggtctccg ctggtggtcc
     3661 taccatacat gaaacatgga gatcttcgaa atttcattcg aaatgagact cataatccaa
     3721 ctgtaaaaga tcttattggc tttggtcttc aagtagccaa aggcatgaaa tatcttgcaa
     3781 gcaaaaagtt tgtccacaga gacttggctg caagaaactg tatgctggat gaaaaattca
     3841 cagtcaaggt tgctgatttt ggtcttgcca gagacatgta tgataaagaa tactatagtg
     3901 tacacaacaa aacaggtgca aagctgccag tgaagtggat ggctttggaa agtctgcaaa
     3961 ctcaaaagtt taccaccaag tcagatgtgt ggtcctttgg cgtgctcctc tgggagctga
     4021 tgacaagagg agccccacct tatcctgacg taaacacctt tgatataact gtttacttgt
     4081 tgcaagggag aagactccta caacccgaat actgcccaga ccccttatat gaagtaatgc
     4141 taaaatgctg gcaccctaaa gccgaaatgc gcccatcctt ttctgaactg gtgtcccgga
     4201 tatcagcgat cttctctact ttcattgggg agcactatgt ccatgtgaac gctacttatg
     4261 tgaacgtaaa atgtgtcgct ccgtatcctt ctctgttgtc atcagaagat aacgctgatg
     4321 atgaggtgga cacacgacca gcctccttct gggagacatc accaactttc ttgtacaaag
     4381 ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc
     4441 aaaataaaat cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg
     4501 tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac
     4561 aagataaaaa tatatcatca tgcctcctct agaccagcca ggacagaaat gcctcgactt
     4621 cgctgctacc caaggttgcc gggtgacgca caccgtggaa acggatgaag gcacgaaccc
     4681 agtggacata agcctgttcg gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca
     4741 actggtccag aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg
     4801 cttgttatga ctgttttttt ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc
     4861 gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagggca
     4921 gtcgccctaa aacaaagtta aacattatga gggaagcggt gatcgccgaa gtatcgactc
     4981 aactatcaga ggtagttggc gtcatcgagc gccatctcga accgacgttg ctggccgtac
     5041 atttgtacgg ctccgcagtg gatggcggcc tgaagccaca cagtgatatt gatttgctgg
     5101 ttacggtgac cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg
     5161 aaacttcggc ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc accattgttg
     5221 tgcacgacga catcattccg tggcgttatc cagctaagcg cgaactgcaa tttggagaat
     5281 ggcagcgcaa tgacattctt gcaggtatct tcgagccagc cacgatcgac attgatctgg
     5341 ctatcttgct gacaaaagca agagaacata gcgttgcctt ggtaggtcca gcggcggagg
     5401 aactctttga tccggttcct gaacaggatc tatttgaggc gctaaatgaa accttaacgc
     5461 tatggaactc gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc
     5521 gcatttggta cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg
     5581 caatggagcg cctgccggcc cagtatcagc ccgtcatact tgaagctaga caggcttatc
     5641 ttggacaaga agaagatcgc ttggcctcgc gcgcagatca gttggaagaa tttgtccact
     5701 acgtgaaagg cgagatcacc aaggtagtcg gcaaataacc ctcgagccac ccatgaccaa
     5761 aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca
     5821 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5881 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5941 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     6001 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6061 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6121 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6181 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6241 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6301 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6361 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6421 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     6481 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     6541 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     6601 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     6661 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg
     6721 ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt
     6781 agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt
     6841 cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa
     6901 acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct
//
