LOCUS       sequence-167969-        3993 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 3993)
  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 3993)
  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_p.S184L
FEATURES             Location/Qualifiers
     source          1..3993
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      25..613
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     514..533
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     767..784
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(943..970)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1062..1148)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1197..1219
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1211..1228
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1212..1228
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1244..1343
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      1344..1353
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2547..2646)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2663..2682)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2664..2682)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2687..2703)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2687..2703)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3134..3925
                     /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"
ORIGIN
        1 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
       61 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
      121 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
      181 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
      241 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
      301 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
      361 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
      421 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
      481 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
      541 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
      601 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
      661 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
      721 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
      781 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
      841 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
      901 taatacgcgt accgctagcc aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg
      961 atggccttct gcttagtttg atgcctggca gtttatggcg ggcgtcctgc ccgccaccct
     1021 ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg gatttgtcct actcaggaga
     1081 gcgttcaccg acaaacaaca gataaaacga aaggcccagt cttccgactg agcctttcgt
     1141 tttatttgat gcctggcagt tccctactct cgcgttaacg ctagcatgga tgttttccca
     1201 gtcacgacgt tgtaaaacga cggccagtct taagctcggg ccccaaataa tgattttatt
     1261 ttgactgata gtgacctgtt cgttgcaaca aattgatgag caatgctttt ttataatgcc
     1321 aactttgtac aaaaaagttg gcaccatggc ggctccgagc ccgagcggcg gcggcggctc
     1381 cgggggcggc agcggcagcg gcacccccgg ccccgtaggg tccccggcgc caggccaccc
     1441 ggccgtcagc agcatgcagg gtaaacgcaa agcactgaag ttgaattttg caaatccacc
     1501 tttcaaatct acagcaaggt ttactctgaa tcccaatcct acaggagttc aaaacccaca
     1561 catagagaga ctgagaacac acagcattga gtcatcagga aaactgaaga tctcccctga
     1621 acaacactgg gatttcactg cagaggactt gaaagacctt ggagaaattg gacgaggagc
     1681 ttatggttct gtcaacaaaa tggtccacaa accaagtggg caaataatgg cagttaaaag
     1741 aattcggtca acagtggatg aaaaagaaca aaaacaactt cttatggatt tggatgtagt
     1801 aatgcggagt agtgattgcc catacattgt tcagttttat ggtgcactct tcagagaggg
     1861 tgactgttgg atctgtatgg aactcatgtc taccttgttt gataagtttt acaaatatgt
     1921 atatagtgta ttagatgatg ttattccaga agaaatttta ggcaaaatca ctttagcaac
     1981 tgtgaaagca ctaaaccact taaaagaaaa cttgaaaatt attcacagag atatcaaacc
     2041 ttccaatatt cttctggaca gaagtggaaa tattaagctc tgtgacttcg gcatcagtgg
     2101 acagcttgtg gactctattg ccaagacaag agatgctggc tgtaggccat acatggcacc
     2161 tgaaagaata gacccaagcg catcacgaca aggatatgat gtccgctctg atgtctggag
     2221 tttggggatc acattgtatg agttggccac aggccgattt ccttatccaa agtggaatag
     2281 tgtatttgat caactaacac aagtcgtgaa aggagatcct ccgcagctga gtaattctga
     2341 ggaaagggaa ttctccccga gtttcatcaa ctttgtcaac ttgtgcctta cgaaggatga
     2401 atccaaaagg ccaaagtata aagagcttct gaaacatccc tttattttga tgtatgaaga
     2461 acgtgccgtt gaggtcgcat gctatgtttg taaaatcctg gatcaaatgc cagctactcc
     2521 cagctctccc atgtatgtcg attgattgcc aactttcttg tacaaagttg gcattataag
     2581 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
     2641 tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc
     2701 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     2761 atcatcatgc ctcctctaga ccagccagga cagaaatgcc tcgacttcgc tgctacccaa
     2821 ggttgccggg tgacgcacac cgtggaaacg gatgaaggca cgaacccagt ggacataagc
     2881 ctgttcggtt cgtaagctgt aatgcaagta gcgtatgcgc tcacgcaact ggtccagaac
     2941 cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt gttatgactg
     3001 tttttttggg gtacagtcta tgcctcgggc atccaagcag caagcgcgtt acgccgtggg
     3061 tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagggcagtc gccctaaaac
     3121 aaagttaaac attatgaggg aagcggtgat cgccgaagta tcgactcaac tatcagaggt
     3181 agttggcgtc atcgagcgcc atctcgaacc gacgttgctg gccgtacatt tgtacggctc
     3241 cgcagtggat ggcggcctga agccacacag tgatattgat ttgctggtta cggtgaccgt
     3301 aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa cttcggcttc
     3361 ccctggagag agcgagattc tccgcgctgt agaagtcacc attgttgtgc acgacgacat
     3421 cattccgtgg cgttatccag ctaagcgcga actgcaattt ggagaatggc agcgcaatga
     3481 cattcttgca ggtatcttcg agccagccac gatcgacatt gatctggcta tcttgctgac
     3541 aaaagcaaga gaacatagcg ttgccttggt aggtccagcg gcggaggaac tctttgatcc
     3601 ggttcctgaa caggatctat ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc
     3661 gcccgactgg gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca tttggtacag
     3721 cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa tggagcgcct
     3781 gccggcccag tatcagcccg tcatacttga agctagacag gcttatcttg gacaagaaga
     3841 agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gtccactacg tgaaaggcga
     3901 gatcaccaag gtagtcggca aataaccctc gagccaccca tgaccaaaat cccttaacgt
     3961 gagttacgcg tcgttccact gagcgtcaga ccc
//
