LOCUS       sequence-167935-        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.G12A
FEATURES             Location/Qualifiers
     source          1..3360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      17..605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     506..525
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     759..776
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(935..962)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1054..1140)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1189..1211
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1203..1220
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1204..1220
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1236..1335
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(1338..1370,1371..1373,1374..1904)
                     /codon_start=1
                     /product="human oncoprotein generated by the G12V mutation 
                     in the small GTPase K-Ras"
                     /label=K-Ras (G12V)
                     /translation="MTEYKLVVVGAAGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
                     DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
                     RKHKEKMSKDGKKKKKKSKTKCVIM"
     CDS             1338..1904
                     /codon_start=1
                     /product="human small GTPase proto-oncoprotein"
                     /label=K-Ras
                     /translation="MTEYKLVVVGAAGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
                     DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
                     RKHKEKMSKDGKKKKKKSKTKCVIM"
     protein_bind    complement(1906..2005)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2022..2041)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2023..2041)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2046..2062)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2046..2062)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2493..3284
                     /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 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
       61 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      121 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      181 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      241 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      301 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      361 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      421 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      481 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      541 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      601 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      661 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
      721 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
      781 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
      841 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatacgc
      901 gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt
      961 ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg
     1021 ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga gagcgttcac
     1081 cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc gttttatttg
     1141 atgcctggca gttccctact ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac
     1201 gttgtaaaac gacggccagt cttaagctcg ggccccaaat aatgatttta ttttgactga
     1261 tagtgacctg ttcgttgcaa caaattgatg agcaatgctt ttttataatg ccaactttgt
     1321 acaaaaaagt tggcaccatg actgaatata aacttgtggt agttggagct gctggcgtag
     1381 gcaagagtgc cttgacgata cagctaattc agaatcattt tgtggacgaa tatgatccaa
     1441 caatagagga ttcctacagg aagcaagtag taattgatgg agaaacctgt ctcttggata
     1501 ttctcgacac agcaggtcaa gaggagtaca gtgcaatgag ggaccagtac atgaggactg
     1561 gggagggctt tctttgtgta tttgccataa ataatactaa atcatttgaa gatattcacc
     1621 attatagaga acaaattaaa agagttaagg actctgaaga tgtacctatg gtcctagtag
     1681 gaaataaatg tgatttgcct tctagaacag tagacacaaa acaggctcag gacttagcaa
     1741 gaagttatgg aattcctttt attgaaacat cagcaaagac aagacagggt gttgatgatg
     1801 ccttctatac attagttcga gaaattcgaa aacataaaga aaagatgagc aaagatggta
     1861 aaaagaagaa aaagaagtca aagacaaagt gtgtaattat gtaattgcca actttcttgt
     1921 acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta
     1981 tcagtcaaaa taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt
     2041 acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca
     2101 ttgcacaaga taaaaatata tcatcatgcc tcctctagac cagccaggac agaaatgcct
     2161 cgacttcgct gctacccaag gttgccgggt gacgcacacc gtggaaacgg atgaaggcac
     2221 gaacccagtg gacataagcc tgttcggttc gtaagctgta atgcaagtag cgtatgcgct
     2281 cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt
     2341 tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc
     2401 aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc
     2461 agggcagtcg ccctaaaaca aagttaaaca ttatgaggga agcggtgatc gccgaagtat
     2521 cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg
     2581 ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt
     2641 tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc
     2701 ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca
     2761 ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg
     2821 gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg
     2881 atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg
     2941 cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct
     3001 taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt
     3061 tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg
     3121 actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg
     3181 cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg
     3241 tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg agccacccat
     3301 gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa
//
