LOCUS       sequence-165278-        4734 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing RAF1 , part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4734)
  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 4734)
  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_RAF1_WT
FEATURES             Location/Qualifiers
     source          1..4734
                     /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(2134..2233)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2250..2269)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2251..2269)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2274..2290)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2274..2290)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2721..3512
                     /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      3605..4193
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4094..4113
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4347..4364
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4523..4550)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4642..4728)
                     /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 aagttggcat ggagcacata cagggagctt ggaagacgat cagcaatggt tttggattca
      241 aagatgccgt gtttgatggc tccagctgca tctctcctac aatagttcag cagtttggct
      301 atcagcgccg ggcatcagat gatggcaaac tcacagatcc ttctaagaca agcaacacta
      361 tccgtgtttt cttgccgaac aagcaaagaa cagtggtcaa tgtgcgaaat ggaatgagct
      421 tgcatgactg ccttatgaaa gcactcaagg tgaggggcct gcaaccagag tgctgtgcag
      481 tgttcagact tctccacgaa cacaaaggta aaaaagcacg cttagattgg aatactgatg
      541 ctgcgtcttt gattggagaa gaacttcaag tagatttcct ggatcatgtt cccctcacaa
      601 cacacaactt tgctcggaag acgttcctga agcttgcctt ctgtgacatc tgtcagaaat
      661 tcctgctcaa tggatttcga tgtcagactt gtggctacaa atttcatgag cactgtagca
      721 ccaaagtacc tactatgtgt gtggactgga gtaacatcag acaactctta ttgtttccaa
      781 attccactat tggtgatagt ggagtcccag cactaccttc tttgactatg cgtcgtatgc
      841 gagagtctgt ttccaggatg cctgttagtt ctcagcacag atattctaca cctcacgcct
      901 tcacctttaa cacctccagt ccctcatctg aaggttccct ctcccagagg cagaggtcga
      961 catccacacc taatgtccac atggtcagca ccaccctgcc tgtggacagc aggatgattg
     1021 aggatgcaat tcgaagtcac agcgaatcag cctcaccttc agccctgtcc agtagcccca
     1081 acaatctgag cccaacaggc tggtcacagc cgaaaacccc cgtgccagca caaagagagc
     1141 gggcaccagt atctgggacc caggagaaaa acaaaattag gcctcgtgga cagagagatt
     1201 caagctatta ttgggaaata gaagccagtg aagtgatgct gtccactcgg attgggtcag
     1261 gctcttttgg aactgtttat aagggtaaat ggcacggaga tgttgcagta aagatcctaa
     1321 aggttgtcga cccaacccca gagcaattcc aggccttcag gaatgaggtg gctgttctgc
     1381 gcaaaacacg gcatgtgaac attctgcttt tcatggggta catgacaaag gacaacctgg
     1441 caattgtgac ccagtggtgc gagggcagca gcctctacaa acacctgcat gtccaggaga
     1501 ccaagtttca gatgttccag ctaattgaca ttgcccggca gacggctcag ggaatggact
     1561 atttgcatgc aaagaacatc atccatagag acatgaaatc caacaatata tttctccatg
     1621 aaggcttaac agtgaaaatt ggagattttg gtttggcaac agtaaagtca cgctggagtg
     1681 gttctcagca ggttgaacaa cctactggct ctgtcctctg gatggcccca gaggtgatcc
     1741 gaatgcagga taacaaccca ttcagtttcc agtcggatgt ctactcctat ggcatcgtat
     1801 tgtatgaact gatgacgggg gagcttcctt attctcacat caacaaccga gatcagatca
     1861 tcttcatggt gggccgagga tatgcctccc cagatcttag taagctatat aagaactgcc
     1921 ccaaagcaat gaagaggctg gtagctgact gtgtgaagaa agtaaaggaa gagaggcctc
     1981 tttttcccca gatcctgtct tccattgagc tgctccaaca ctctctaccg aagatcaacc
     2041 ggagcgcttc cgagccatcc ttgcatcggg cagcccacac tgaggatatc aatgcttgca
     2101 cgctgaccac gtccccgagg ctgcctgtct tctacccaac tttcttgtac aaagttggca
     2161 ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata
     2221 aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag
     2281 ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata
     2341 aaaatatatc atcatgcctc ctctagacca gccaggacag aaatgcctcg acttcgctgc
     2401 tacccaaggt tgccgggtga cgcacaccgt ggaaacggat gaaggcacga acccagtgga
     2461 cataagcctg ttcggttcgt aagctgtaat gcaagtagcg tatgcgctca cgcaactggt
     2521 ccagaacctt gaccgaacgc agcggtggta acggcgcagt ggcggttttc atggcttgtt
     2581 atgactgttt ttttggggta cagtctatgc ctcgggcatc caagcagcaa gcgcgttacg
     2641 ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt tacgcagcag ggcagtcgcc
     2701 ctaaaacaaa gttaaacatt atgagggaag cggtgatcgc cgaagtatcg actcaactat
     2761 cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc gtacatttgt
     2821 acggctccgc agtggatggc ggcctgaagc cacacagtga tattgatttg ctggttacgg
     2881 tgaccgtaag gcttgatgaa acaacgcggc gagctttgat caacgacctt ttggaaactt
     2941 cggcttcccc tggagagagc gagattctcc gcgctgtaga agtcaccatt gttgtgcacg
     3001 acgacatcat tccgtggcgt tatccagcta agcgcgaact gcaatttgga gaatggcagc
     3061 gcaatgacat tcttgcaggt atcttcgagc cagccacgat cgacattgat ctggctatct
     3121 tgctgacaaa agcaagagaa catagcgttg ccttggtagg tccagcggcg gaggaactct
     3181 ttgatccggt tcctgaacag gatctatttg aggcgctaaa tgaaacctta acgctatgga
     3241 actcgccgcc cgactgggct ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt
     3301 ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg
     3361 agcgcctgcc ggcccagtat cagcccgtca tacttgaagc tagacaggct tatcttggac
     3421 aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc cactacgtga
     3481 aaggcgagat caccaaggta gtcggcaaat aaccctcgag ccacccatga ccaaaatccc
     3541 ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3601 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3661 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3721 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     3781 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3841 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3901 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3961 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4021 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4081 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4141 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4201 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     4261 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     4321 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     4381 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     4441 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt accgctagcc
     4501 aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttagtttg
     4561 atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac
     4621 gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
     4681 gataaaacga aaggcccagt cttccgactg agcctttcgt tttatttgat gcct
//
