LOCUS       sequence-163162-        3876 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing MAPK1 , part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3876)
  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 3876)
  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_MAPK1_WT
FEATURES             Location/Qualifiers
     source          1..3876
                     /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"
     CDS             1236..1271
                     /codon_start=1
                     /product="epitope tag derived from MAP kinase 2"
                     /label=Tag-100
                     /translation="EETARFQPGYRS"
     protein_bind    complement(1276..1375)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1392..1411)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1393..1411)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1416..1432)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1416..1432)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1863..2654
                     /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      2747..3335
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3236..3255
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3489..3506
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3665..3692)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3784..3870)
                     /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 catggcggcg gcggcggcgg cgggcgcggg cccggagatg gtccgcgggc
      241 aggtgttcga cgtggggccg cgctacacca acctctcgta catcggcgag ggcgcctacg
      301 gcatggtgtg ctctgcttat gataatgtca acaaagttcg agtagctatc aagaaaatca
      361 gcccctttga gcaccagacc tactgccaga gaaccctgag ggagataaaa atcttactgc
      421 gcttcagaca tgagaacatc attggaatca atgacattat tcgagcacca accatcgagc
      481 aaatgaaaga tgtatatata gtacaggacc tcatggaaac agatctttac aagctcttga
      541 agacacaaca cctcagcaat gaccatatct gctattttct ctaccagatc ctcagagggt
      601 taaaatatat ccattcagct aacgttctgc accgtgacct caagccttcc aacctgctgc
      661 tcaacaccac ctgtgatctc aagatctgtg actttggcct ggcccgtgtt gcagatccag
      721 accatgatca cacagggttc ctgacagaat atgtggccac acgttggtac agggctccag
      781 aaattatgtt gaattccaag ggctacacca agtccattga tatttggtct gtaggctgca
      841 ttctggcaga aatgctttct aacaggccca tctttccagg gaagcattat cttgaccagc
      901 tgaaccacat tttgggtatt cttggatccc catcacaaga agacctgaat tgtataataa
      961 atttaaaagc taggaactat ttgctttctc ttccacacaa aaataaggtg ccatggaaca
     1021 ggctgttccc aaatgctgac tccaaagctc tggacttatt ggacaaaatg ttgacattca
     1081 acccacacaa gaggattgaa gtagaacagg ctctggccca cccatatctg gagcagtatt
     1141 acgacccgag tgacgagccc atcgccgaag caccattcaa gttcgacatg gaattggatg
     1201 acttgcctaa ggaaaagctc aaagaactaa tttttgaaga gactgctaga ttccagccag
     1261 gatacagatc ttagttgcca actttcttgt acaaagttgg cattataaga aagcattgct
     1321 tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc
     1381 cagctgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct
     1441 ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgcc
     1501 tcctctagac cagccaggac agaaatgcct cgacttcgct gctacccaag gttgccgggt
     1561 gacgcacacc gtggaaacgg atgaaggcac gaacccagtg gacataagcc tgttcggttc
     1621 gtaagctgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac
     1681 gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg
     1741 tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga
     1801 tgttatggag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca
     1861 ttatgaggga agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca
     1921 tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg
     1981 gcggcctgaa gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg
     2041 aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga
     2101 gcgagattct ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc
     2161 gttatccagc taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag
     2221 gtatcttcga gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag
     2281 aacatagcgt tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac
     2341 aggatctatt tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg
     2401 ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg
     2461 gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt
     2521 atcagcccgt catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg
     2581 cctcgcgcgc agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg
     2641 tagtcggcaa ataaccctcg agccacccat gaccaaaatc ccttaacgtg agttacgcgt
     2701 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     2761 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     2821 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     2881 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     2941 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3001 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3061 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3121 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3181 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     3241 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     3301 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3361 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     3421 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     3481 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     3541 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     3601 cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa
     3661 cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg
     3721 cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg
     3781 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
     3841 gtcttccgac tgagcctttc gttttatttg atgcct
//
