LOCUS       sequence-168829-        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_WT
FEATURES             Location/Qualifiers
     source          1..3360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     356..375
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     609..626
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(785..812)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(904..990)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1039..1061
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1053..1070
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1054..1070
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1086..1185
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1188..1754
                     /codon_start=1
                     /product="human small GTPase proto-oncoprotein"
                     /label=K-Ras
                     /translation="MTEYKLVVVGAGGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
                     DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
                     RKHKEKMSKDGKKKKKKSKTKCVIM"
     protein_bind    complement(1756..1855)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1872..1891)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1873..1891)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1896..1912)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1896..1912)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2343..3134
                     /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      join(3227..3360,1..455)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
       61 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
      121 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
      181 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
      241 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
      301 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
      361 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
      421 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
      481 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
      541 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
      601 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
      661 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
      721 cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa
      781 cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg
      841 cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg
      901 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
      961 gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa
     1021 cgctagcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt cttaagctcg
     1081 ggccccaaat aatgatttta ttttgactga tagtgacctg ttcgttgcaa caaattgatg
     1141 agcaatgctt ttttataatg ccaactttgt acaaaaaagt tggcaccatg actgaatata
     1201 aacttgtggt agttggagct ggtggcgtag gcaagagtgc cttgacgata cagctaattc
     1261 agaatcattt tgtggacgaa tatgatccaa caatagagga ttcctacagg aagcaagtag
     1321 taattgatgg agaaacctgt ctcttggata ttctcgacac agcaggtcaa gaggagtaca
     1381 gtgcaatgag ggaccagtac atgaggactg gggagggctt tctttgtgta tttgccataa
     1441 ataatactaa atcatttgaa gatattcacc attatagaga acaaattaaa agagttaagg
     1501 actctgaaga tgtacctatg gtcctagtag gaaataaatg tgatttgcct tctagaacag
     1561 tagacacaaa acaggctcag gacttagcaa gaagttatgg aattcctttt attgaaacat
     1621 cagcaaagac aagacagggt gttgatgatg ccttctatac attagttcga gaaattcgaa
     1681 aacataaaga aaagatgagc aaagatggta aaaagaagaa aaagaagtca aagacaaagt
     1741 gtgtaattat gtgattgcca actttcttgt acaaagttgg cattataaga aagcattgct
     1801 tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc
     1861 cagctgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct
     1921 ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgcc
     1981 tcctctagac cagccaggac agaaatgcct cgacttcgct gctacccaag gttgccgggt
     2041 gacgcacacc gtggaaacgg atgaaggcac gaacccagtg gacataagcc tgttcggttc
     2101 gtaagctgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac
     2161 gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg
     2221 tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga
     2281 tgttatggag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca
     2341 ttatgaggga agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca
     2401 tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg
     2461 gcggcctgaa gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg
     2521 aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga
     2581 gcgagattct ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc
     2641 gttatccagc taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag
     2701 gtatcttcga gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag
     2761 aacatagcgt tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac
     2821 aggatctatt tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg
     2881 ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg
     2941 gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt
     3001 atcagcccgt catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg
     3061 cctcgcgcgc agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg
     3121 tagtcggcaa ataaccctcg agccacccat gaccaaaatc ccttaacgtg agttacgcgt
     3181 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3241 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3301 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
//
