LOCUS       sequence-169335-        3975 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing MAP2K1 , part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3975)
  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 3975)
  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_MAP2K1_WT
FEATURES             Location/Qualifiers
     source          1..3975
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1854..1953)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1970..1989)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1971..1989)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1994..2010)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1994..2010)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2441..3232
                     /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      3325..3913
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3814..3833
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agttggcacc atgcccaaga agaagccgac gcccatccag ctgaacccgg cccccgacgg
      721 ctctgcagtt aacgggacca gctctgcgga gaccaacttg gaggccttgc agaagaagct
      781 ggaggagcta gagcttgatg agcagcagcg aaagcgcctt gaggcctttc ttacccagaa
      841 gcagaaggtg ggagaactga aggatgacga ctttgagaag atcagtgagc tgggggctgg
      901 caatggcggt gtggtgttca aggtctccca caagccttct ggcctggtca tggccagaaa
      961 gctaattcat ctggagatca aacccgcaat ccggaaccag atcataaggg agctgcaggt
     1021 tctgcatgag tgcaactctc cgtacatcgt gggcttctat ggtgcgttct acagcgatgg
     1081 cgagatcagt atctgcatgg agcacatgga tggaggttct ctggatcaag tcctgaagaa
     1141 agctggaaga attcctgaac aaattttagg aaaagttagc attgctgtaa taaaaggcct
     1201 gacatatctg agggagaagc acaagatcat gcacagagat gtcaagccct ccaacatcct
     1261 agtcaactcc cgtggggaga tcaagctctg tgactttggg gtcagcgggc agctcatcga
     1321 ctccatggcc aactccttcg tgggcacaag gtcctacatg tcgccagaaa gactccaggg
     1381 gactcattac tctgtgcagt cagacatctg gagcatggga ctgtctctgg tagagatggc
     1441 ggttgggagg tatcccatcc ctcctccaga tgccaaggag ctggagctga tgtttgggtg
     1501 ccaggtggaa ggagatgcgg ctgagacccc acccaggcca aggacccccg ggaggcccct
     1561 tagctcatac ggaatggaca gccgacctcc catggcaaat tttgagttgt tggattacat
     1621 agtcaacgag cctcctccaa aactgcccag tggagtgttc agtctggaat ttcaagattt
     1681 tgtgaataaa tgcttaataa aaaaccccgc agagagagca gatttgaagc aactcatggt
     1741 tcatgctttt atcaagagat ctgatgctga ggaagtggat tttgcaggtt ggctctgctc
     1801 caccatcggc cttaaccagc ccagcacacc aacccatgct gctggcgtct gattgccaac
     1861 tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca
     1921 ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga
     1981 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
     2041 atgttacatt gcacaagata aaaatatatc atcatgcctc ctctagacca gccaggacag
     2101 aaatgcctcg acttcgctgc tacccaaggt tgccgggtga cgcacaccgt ggaaacggat
     2161 gaaggcacga acccagtgga cataagcctg ttcggttcgt aagctgtaat gcaagtagcg
     2221 tatgcgctca cgcaactggt ccagaacctt gaccgaacgc agcggtggta acggcgcagt
     2281 ggcggttttc atggcttgtt atgactgttt ttttggggta cagtctatgc ctcgggcatc
     2341 caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt
     2401 tacgcagcag ggcagtcgcc ctaaaacaaa gttaaacatt atgagggaag cggtgatcgc
     2461 cgaagtatcg actcaactat cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac
     2521 gttgctggcc gtacatttgt acggctccgc agtggatggc ggcctgaagc cacacagtga
     2581 tattgatttg ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc gagctttgat
     2641 caacgacctt ttggaaactt cggcttcccc tggagagagc gagattctcc gcgctgtaga
     2701 agtcaccatt gttgtgcacg acgacatcat tccgtggcgt tatccagcta agcgcgaact
     2761 gcaatttgga gaatggcagc gcaatgacat tcttgcaggt atcttcgagc cagccacgat
     2821 cgacattgat ctggctatct tgctgacaaa agcaagagaa catagcgttg ccttggtagg
     2881 tccagcggcg gaggaactct ttgatccggt tcctgaacag gatctatttg aggcgctaaa
     2941 tgaaacctta acgctatgga actcgccgcc cgactgggct ggcgatgagc gaaatgtagt
     3001 gcttacgttg tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt
     3061 cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca tacttgaagc
     3121 tagacaggct tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga
     3181 agaatttgtc cactacgtga aaggcgagat caccaaggta gtcggcaaat aaccctcgag
     3241 ccacccatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc
     3301 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     3361 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     3421 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     3481 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3541 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3601 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3661 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3721 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3781 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3841 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3901 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3961 ttttgctcac atgtt
//
