LOCUS       sequence-166545-        3360 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing KRAS,part of the Target Accelerator 
            Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3360)
  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 3360)
  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_KRAS_p.G12V
FEATURES             Location/Qualifiers
     source          1..3360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      18..606
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     507..526
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     760..777
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(936..963)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1055..1141)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1190..1212
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1204..1221
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1205..1221
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1237..1336
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(1339..1371,1372..1374,1375..1905)
                     /codon_start=1
                     /product="human oncoprotein generated by the G12V mutation 
                     in the small GTPase K-Ras"
                     /label=K-Ras (G12V)
                     /translation="MTEYKLVVVGAVGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
                     DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
                     RKHKEKMSKDGKKKKKKSKTKCVIM"
     protein_bind    complement(1907..2006)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2023..2042)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2024..2042)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2047..2063)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2047..2063)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2494..3285
                     /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 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
       61 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
      121 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
      181 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
      241 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
      301 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
      361 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
      421 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
      481 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
      541 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
      601 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
      661 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
      721 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
      781 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
      841 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg
      901 cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct
      961 tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc
     1021 gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca
     1081 ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt
     1141 gatgcctggc agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga
     1201 cgttgtaaaa cgacggccag tcttaagctc gggccccaaa taatgatttt attttgactg
     1261 atagtgacct gttcgttgca acaaattgat gagcaatgct tttttataat gccaactttg
     1321 tacaaaaaag ttggcaccat gactgaatat aaacttgtgg tagttggagc tgttggcgta
     1381 ggcaagagtg ccttgacgat acagctaatt cagaatcatt ttgtggacga atatgatcca
     1441 acaatagagg attcctacag gaagcaagta gtaattgatg gagaaacctg tctcttggat
     1501 attctcgaca cagcaggtca agaggagtac agtgcaatga gggaccagta catgaggact
     1561 ggggagggct ttctttgtgt atttgccata aataatacta aatcatttga agatattcac
     1621 cattatagag aacaaattaa aagagttaag gactctgaag atgtacctat ggtcctagta
     1681 ggaaataaat gtgatttgcc ttctagaaca gtagacacaa aacaggctca ggacttagca
     1741 agaagttatg gaattccttt tattgaaaca tcagcaaaga caagacaggg tgttgatgat
     1801 gccttctata cattagttcg agaaattcga aaacataaag aaaagatgag caaagatggt
     1861 aaaaagaaga aaaagaagtc aaagacaaag tgtgtaatta tgtaattgcc aactttcttg
     1921 tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact
     1981 atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat
     2041 tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac
     2101 attgcacaag ataaaaatat atcatcatgc ctcctctaga ccagccagga cagaaatgcc
     2161 tcgacttcgc tgctacccaa ggttgccggg tgacgcacac cgtggaaacg gatgaaggca
     2221 cgaacccagt ggacataagc ctgttcggtt cgtaagctgt aatgcaagta gcgtatgcgc
     2281 tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt
     2341 ttcatggctt gttatgactg tttttttggg gtacagtcta tgcctcgggc atccaagcag
     2401 caagcgcgtt acgccgtggg tcgatgtttg atgttatgga gcagcaacga tgttacgcag
     2461 cagggcagtc gccctaaaac aaagttaaac attatgaggg aagcggtgat cgccgaagta
     2521 tcgactcaac tatcagaggt agttggcgtc atcgagcgcc atctcgaacc gacgttgctg
     2581 gccgtacatt tgtacggctc cgcagtggat ggcggcctga agccacacag tgatattgat
     2641 ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac
     2701 cttttggaaa cttcggcttc ccctggagag agcgagattc tccgcgctgt agaagtcacc
     2761 attgttgtgc acgacgacat cattccgtgg cgttatccag ctaagcgcga actgcaattt
     2821 ggagaatggc agcgcaatga cattcttgca ggtatcttcg agccagccac gatcgacatt
     2881 gatctggcta tcttgctgac aaaagcaaga gaacatagcg ttgccttggt aggtccagcg
     2941 gcggaggaac tctttgatcc ggttcctgaa caggatctat ttgaggcgct aaatgaaacc
     3001 ttaacgctat ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt agtgcttacg
     3061 ttgtcccgca tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc
     3121 gactgggcaa tggagcgcct gccggcccag tatcagcccg tcatacttga agctagacag
     3181 gcttatcttg gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt
     3241 gtccactacg tgaaaggcga gatcaccaag gtagtcggca aataaccctc gagccaccca
     3301 tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa
//
