LOCUS       sequence-166522-        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.A59G
FEATURES             Location/Qualifiers
     source          1..3360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      7..595
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     496..515
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     749..766
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(925..952)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1044..1130)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1179..1201
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1193..1210
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1194..1210
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1226..1325
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1328..1894
                     /codon_start=1
                     /product="human small GTPase proto-oncoprotein"
                     /label=K-Ras
                     /translation="MTEYKLVVVGAGGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTGGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
                     DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
                     RKHKEKMSKDGKKKKKKSKTKCVIM"
     protein_bind    complement(1896..1995)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2012..2031)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2013..2031)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2036..2052)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2036..2052)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2483..3274
                     /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 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
       61 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
      121 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
      181 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
      241 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
      301 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
      361 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
      421 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
      481 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
      541 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
      601 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
      661 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
      721 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
      781 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
      841 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatacgc gtaccgctag
      901 ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt
      961 tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca
     1021 acgttcaaat ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa
     1081 cagataaaac gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca
     1141 gttccctact ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac gttgtaaaac
     1201 gacggccagt cttaagctcg ggccccaaat aatgatttta ttttgactga tagtgacctg
     1261 ttcgttgcaa caaattgatg agcaatgctt ttttataatg acaactttgt acaaaaaagt
     1321 tggcaccatg actgaatata aacttgtggt agttggagct ggtggcgtag gcaagagtgc
     1381 cttgacgata cagctaattc agaatcattt tgtggacgaa tatgatccaa caatagagga
     1441 ttcctacagg aagcaagtag taattgatgg agaaacctgt ctcttggata ttctcgacac
     1501 aggaggtcaa gaggagtaca gtgcaatgag ggaccagtac atgaggactg gggagggctt
     1561 tctttgtgta tttgccataa ataatactaa atcatttgaa gatattcacc attatagaga
     1621 acaaattaaa agagttaagg actctgaaga tgtacctatg gtcctagtag gaaataaatg
     1681 tgatttgcct tctagaacag tagacacaaa acaggctcag gacttagcaa gaagttatgg
     1741 aattcctttt attgaaacat cagcaaagac aagacagggt gttgatgatg ccttctatac
     1801 attagttcga gaaattcgaa aacataaaga aaagatgagc aaagatggta aaaagaagaa
     1861 aaagaagtca aagacaaagt gtgtaattat gtaattgcca actttcttgt acaaagttgg
     1921 cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa
     1981 taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat
     2041 agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     2101 taaaaatata tcatcatgcc tcctctagac cagccaggac agaaatgcct cgacttcgct
     2161 gctacccaag gttgccgggt gacgcacacc gtggaaacgg atgaaggcac gaacccagtg
     2221 gacataagcc tgttcggttc gtaagctgta atgcaagtag cgtatgcgct cacgcaactg
     2281 gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt tcatggcttg
     2341 ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc aagcgcgtta
     2401 cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc agggcagtcg
     2461 ccctaaaaca aagttaaaca ttatgaggga agcggtgatc gccgaagtat cgactcaact
     2521 atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt
     2581 gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt tgctggttac
     2641 ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc ttttggaaac
     2701 ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca ttgttgtgca
     2761 cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg gagaatggca
     2821 gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg atctggctat
     2881 cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg cggaggaact
     2941 ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct taacgctatg
     3001 gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat
     3061 ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat
     3121 ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg cttatcttgg
     3181 acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg tccactacgt
     3241 gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg agccacccat gaccaaaatc
     3301 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
//
