LOCUS       sequence-163125-        3990 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing MAP2K4 , part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3990)
  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 3990)
  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_MAP2K4_WT
FEATURES             Location/Qualifiers
     source          1..3990
                     /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"
     regulatory      190..199
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1390..1489)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1506..1525)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1507..1525)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1530..1546)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1530..1546)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1977..2768
                     /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      2861..3449
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3350..3369
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3603..3620
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3779..3806)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3898..3984)
                     /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 catggcggct ccgagcccga gcggcggcgg ctccgggggc ggcagcggca
      241 gcggcacccc cggccccgta gggtcccggg cgccaggcca cccggccgtc agcagcatgc
      301 agggtaaacg caaagcactg aagttgaatt ttgcaaatcc acctttcaaa tctacagcaa
      361 ggtttactct gaatcccaat cctacaggag ttcaaaaccc acacatagag agactgagaa
      421 cacacagcat tgagtcatca ggaaaactga agatctcccc tgaacaacac tgggatttca
      481 ctgcagagga cttgaaagac cttggagaaa ttggacgagg agcttatggt tctgtcaaca
      541 aaatggtcca caaaccaagt gggcaaataa tggcagttaa aagaattcgg tcaacagtgg
      601 atgaaaaaga acaaaaacaa cttcttatgg atttggatgt agtaatgcgg agtagtgatt
      661 gcccatacat tgttcagttt tatggtgcac tcttcagaga gggtgactgt tggatctgta
      721 tggaactcat gtctacctcg tttgataagt tttacaaata tgtatatagt gtattagatg
      781 atgttattcc agaagaaatt ttaggcaaaa tcactttagc aactgtgaaa gcactaaacc
      841 acttaaaaga aaacttgaaa attattcaca gagatatcaa accttccaat attcttctgg
      901 acagaagtgg aaatattaag ctctgtgact tcggcatcag tggacagctt gtggactcta
      961 ttgccaagac aagagatgct ggctgtaggc catacatggc acctgaaaga atagacccaa
     1021 gcgcatcacg acaaggatat gatgtccgct ctgatgtctg gagtttgggg atcacattgt
     1081 atgagttggc cacaggccga tttccttatc caaagtggaa tagtgtattt gatcaactaa
     1141 cacaagtcgt gaaaggagat cctccgcagc tgagtaattc tgaggaaagg gaattctccc
     1201 cgagtttcat caactttgtc aacttgtgcc ttacgaagga tgaatccaaa aggccaaagt
     1261 ataaagagct tctgaaacat ccctttattt tgatgtatga agaacgtgcc gttgaggtcg
     1321 catgctatgt ttgtaaaatc ctggatcaaa tgccagctac tcccagctct cccatgtatg
     1381 tcgattgatt gccaactttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa
     1441 tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg
     1501 atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg
     1561 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgcctcctct
     1621 agaccagcca ggacagaaat gcctcgactt cgctgctacc caaggttgcc gggtgacgca
     1681 caccgtggaa acggatgaag gcacgaaccc agtggacata agcctgttcg gttcgtaagc
     1741 tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg
     1801 gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt ggggtacagt
     1861 ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat
     1921 ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta aacattatga
     1981 gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc gtcatcgagc
     2041 gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg gatggcggcc
     2101 tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa
     2161 cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga gagagcgaga
     2221 ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg tggcgttatc
     2281 cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt gcaggtatct
     2341 tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca agagaacata
     2401 gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct gaacaggatc
     2461 tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac tgggctggcg
     2521 atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta accggcaaaa
     2581 tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc cagtatcagc
     2641 ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc ttggcctcgc
     2701 gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc aaggtagtcg
     2761 gcaaataacc ctcgagccac ccatgaccaa aatcccttaa cgtgagttac gcgtcgttcc
     2821 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     2881 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     2941 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3001 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3061 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3121 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3181 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3241 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3301 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3361 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3421 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     3481 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     3541 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     3601 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     3661 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     3721 gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta gaaacgcaaa
     3781 aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt atggcgggcg
     3841 tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc ccggcggatt
     3901 tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttc
     3961 cgactgagcc tttcgtttta tttgatgcct
//
