LOCUS       sequence-163898-        3972 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 3972)
  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 3972)
  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_p.S218D_S222D
FEATURES             Location/Qualifiers
     source          1..3972
                     /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(1372..1471)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1488..1507)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1489..1507)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1512..1528)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1512..1528)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1959..2750
                     /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      2843..3431
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3332..3351
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3585..3602
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3761..3788)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3880..3966)
                     /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 catgcccaag aagaagccga cgcccatcca gctgaacccg gcccccgacg
      241 gctctgcagt taacgggacc agctctgcgg agaccaactt ggaggccttg cagaagaagc
      301 tggaggagct agagcttgat gagcagcagc gaaagcgcct tgaggccttt cttacccaga
      361 agcagaaggt gggagaactg aaggatgacg actttgagaa gatcagtgag ctgggggctg
      421 gcaatggcgg tgtggtgttc aaggtctccc acaagccttc tggcctggtc atggccagaa
      481 agctaattca tctggagatc aaacccgcaa tccggaacca gatcataagg gagctgcagg
      541 ttctgcatga gtgcaactct ccgtacatcg tgggcttcta tggtgcgttc tacagcgatg
      601 gcgagatcag tatctgcatg gagcacatgg atggaggttc tctggatcaa gtcctgaaga
      661 aagctggaag aattcctgaa caaattttag gaaaagttag cattgctgta ataaaaggcc
      721 tgacatatct gagggagaag cacaagatca tgcacagaga tgtcaagccc tccaacatcc
      781 tagtcaactc ccgtggggag atcaagctct gtgactttgg ggtcagcggg cagctcatcg
      841 acgacatggc caacgacttc gtgggcacaa ggtcctacat gtcgccagaa agactccagg
      901 ggactcatta ctctgtgcag tcagacatct ggagcatggg actgtctctg gtagagatgg
      961 cggttgggag gtatcccatc cctcctccag atgccaagga gctggagctg atgtttgggt
     1021 gccaggtgga aggagatgcg gctgagaccc cacccaggcc aaggaccccc gggaggcccc
     1081 ttagctcata cggaatggac agccgacctc ccatggcaat ttttgagttg ttggattaca
     1141 tagtcaacga gcctcctcca aaactgccca gtggagtgtt cagtctggaa tttcaagatt
     1201 ttgtgaataa atgcttaata aaaaaccccg cagagagagc agatttgaag caactcatgg
     1261 ttcatgcttt tatcaagaga tctgatgctg aggaagtgga ttttgcaggt tggctctgct
     1321 ccaccatcgg ccttaaccag cccagcacac caacccatgc tgctggcgtc ttgccaactt
     1381 tcttgtacaa agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg
     1441 tcactatcag tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt
     1501 cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat
     1561 gttacattgc acaagataaa aatatatcat catgcctcct ctagaccagc caggacagaa
     1621 atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg cacaccgtgg aaacggatga
     1681 aggcacgaac ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta
     1741 tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg
     1801 cggttttcat ggcttgttat gactgttttt ttggggtaca gtctatgcct cgggcatcca
     1861 agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta
     1921 cgcagcaggg cagtcgccct aaaacaaagt taaacattat gagggaagcg gtgatcgccg
     1981 aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt
     2041 tgctggccgt acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagtgata
     2101 ttgatttgct ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca
     2161 acgacctttt ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag
     2221 tcaccattgt tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc
     2281 aatttggaga atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg
     2341 acattgatct ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc
     2401 cagcggcgga ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg
     2461 aaaccttaac gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc
     2521 ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg
     2581 ctgccgactg ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta
     2641 gacaggctta tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag
     2701 aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa ccctcgagcc
     2761 acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg
     2821 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2881 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2941 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     3001 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3061 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3121 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3181 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     3241 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3301 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     3361 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3421 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3481 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     3541 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     3601 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     3661 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     3721 atacgcgtac cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat
     3781 ggccttctgc ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc
     3841 gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc
     3901 gttcaccgac aaacaacaga taaaacgaaa ggcccagtct tccgactgag cctttcgttt
     3961 tatttgatgc ct
//
