LOCUS       sequence-166475-        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.G12D
FEATURES             Location/Qualifiers
     source          1..3360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      28..616
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     517..536
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     770..787
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(946..973)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1065..1151)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1200..1222
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1214..1231
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1215..1231
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1247..1346
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(1349..1381,1382..1384,1385..1915)
                     /codon_start=1
                     /product="human oncoprotein generated by the G12V mutation 
                     in the small GTPase K-Ras"
                     /label=K-Ras (G12V)
                     /translation="MTEYKLVVVGADGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
                     DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
                     RKHKEKMSKDGKKKKKKSKTKCVIM"
     CDS             1349..1915
                     /codon_start=1
                     /product="human small GTPase proto-oncoprotein"
                     /label=K-Ras
                     /translation="MTEYKLVVVGADGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
                     DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
                     RKHKEKMSKDGKKKKKKSKTKCVIM"
     protein_bind    complement(1917..2016)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2033..2052)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2034..2052)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2057..2073)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2057..2073)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2504..3295
                     /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 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
       61 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
      121 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
      181 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
      241 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
      301 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
      361 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
      421 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
      481 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
      541 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
      601 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
      661 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
      721 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
      781 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
      841 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
      901 aattaatacg cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc
      961 aggatggcct tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac
     1021 cctccgggcc gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg
     1081 agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt
     1141 cgttttattt gatgcctggc agttccctac tctcgcgtta acgctagcat ggatgttttc
     1201 ccagtcacga cgttgtaaaa cgacggccag tcttaagctc gggccccaaa taatgatttt
     1261 attttgactg atagtgacct gttcgttgca acaaattgat gagcaatgct tttttataat
     1321 gacaactttg tacaaaaaag ttggcaccat gactgaatat aaacttgtgg tagttggagc
     1381 tgatggcgta ggcaagagtg ccttgacgat acagctaatt cagaatcatt ttgtggacga
     1441 atatgatcca acaatagagg attcctacag gaagcaagta gtaattgatg gagaaacctg
     1501 tctcttggat attctcgaca cagcaggtca agaggagtac agtgcaatga gggaccagta
     1561 catgaggact ggggagggct ttctttgtgt atttgccata aataatacta aatcatttga
     1621 agatattcac cattatagag aacaaattaa aagagttaag gactctgaag atgtacctat
     1681 ggtcctagta ggaaataaat gtgatttgcc ttctagaaca gtagacacaa aacaggctca
     1741 ggacttagca agaagttatg gaattccttt tattgaaaca tcagcaaaga caagacaggg
     1801 tgttgatgat gccttctata cattagttcg agaaattcga aaacataaag aaaagatgag
     1861 caaagatggt aaaaagaaga aaaagaagtc aaagacaaag tgtgtaatta tgtaattgcc
     1921 aactttcttg tacaaagttg gcattataag aaagcattgc ttatcaattt gttgcaacga
     1981 acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag
     2041 tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct
     2101 ctgatgttac attgcacaag ataaaaatat atcatcatgc ctcctctaga ccagccagga
     2161 cagaaatgcc tcgacttcgc tgctacccaa ggttgccggg tgacgcacac cgtggaaacg
     2221 gatgaaggca cgaacccagt ggacataagc ctgttcggtt cgtaagctgt aatgcaagta
     2281 gcgtatgcgc tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc
     2341 agtggcggtt ttcatggctt gttatgactg tttttttggg gtacagtcta tgcctcgggc
     2401 atccaagcag caagcgcgtt acgccgtggg tcgatgtttg atgttatgga gcagcaacga
     2461 tgttacgcag cagggcagtc gccctaaaac aaagttaaac attatgaggg aagcggtgat
     2521 cgccgaagta tcgactcaac tatcagaggt agttggcgtc atcgagcgcc atctcgaacc
     2581 gacgttgctg gccgtacatt tgtacggctc cgcagtggat ggcggcctga agccacacag
     2641 tgatattgat ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt
     2701 gatcaacgac cttttggaaa cttcggcttc ccctggagag agcgagattc tccgcgctgt
     2761 agaagtcacc attgttgtgc acgacgacat cattccgtgg cgttatccag ctaagcgcga
     2821 actgcaattt ggagaatggc agcgcaatga cattcttgca ggtatcttcg agccagccac
     2881 gatcgacatt gatctggcta tcttgctgac aaaagcaaga gaacatagcg ttgccttggt
     2941 aggtccagcg gcggaggaac tctttgatcc ggttcctgaa caggatctat ttgaggcgct
     3001 aaatgaaacc ttaacgctat ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt
     3061 agtgcttacg ttgtcccgca tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga
     3121 tgtcgctgcc gactgggcaa tggagcgcct gccggcccag tatcagcccg tcatacttga
     3181 agctagacag gcttatcttg gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt
     3241 ggaagaattt gtccactacg tgaaaggcga gatcaccaag gtagtcggca aataaccctc
     3301 gagccaccca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
//
