LOCUS       sequence-166483-        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.G12S
FEATURES             Location/Qualifiers
     source          1..3360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             329..1120
                     /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      1213..1801
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1702..1721
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1955..1972
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2131..2158)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2250..2336)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2385..2407
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2399..2416
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2400..2416
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2432..2531
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2534..3100
                     /codon_start=1
                     /product="human small GTPase proto-oncoprotein"
                     /label=K-Ras
                     /translation="MTEYKLVVVGASGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
                     DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
                     RKHKEKMSKDGKKKKKKSKTKCVIM"
     protein_bind    complement(3102..3201)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3218..3237)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3219..3237)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3242..3258)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3242..3258)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttcgctgcta cccaaggttg ccgggtgacg cacaccgtgg aaacggatga aggcacgaac
       61 ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta tgcgctcacg
      121 caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat
      181 ggcttgttat gactgttttt ttggggtaca gtctatgcct cgggcatcca agcagcaagc
      241 gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcaggg
      301 cagtcgccct aaaacaaagt taaacattat gagggaagcg gtgatcgccg aagtatcgac
      361 tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt
      421 acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagtgata ttgatttgct
      481 ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca acgacctttt
      541 ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag tcaccattgt
      601 tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc aatttggaga
      661 atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg acattgatct
      721 ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc cagcggcgga
      781 ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg aaaccttaac
      841 gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc ttacgttgtc
      901 ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg
      961 ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta gacaggctta
     1021 tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag aatttgtcca
     1081 ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa ccctcgagcc acccatgacc
     1141 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     1201 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     1261 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     1321 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     1381 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     1441 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     1501 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     1561 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     1621 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     1681 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     1741 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     1801 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     1861 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     1921 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     1981 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     2041 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac
     2101 cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc
     2161 ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc
     2221 ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac
     2281 aaacaacaga taaaacgaaa ggcccagtct tccgactgag cctttcgttt tatttgatgc
     2341 ctggcagttc cctactctcg cgttaacgct agcatggatg ttttcccagt cacgacgttg
     2401 taaaacgacg gccagtctta agctcgggcc ccaaataatg attttatttt gactgatagt
     2461 gacctgttcg ttgcaacaaa ttgataagca atgctttttt ataatgacaa ctttgtacaa
     2521 aaaagttggc accatgactg aatataaact tgtggtagtt ggagctagtg gcgtaggcaa
     2581 gagtgccttg acgatacagc taattcagaa tcattttgtg gacgaatatg atccaacaat
     2641 agaggattcc tacaggaagc aagtagtaat tgatggagaa acctgtctct tggatattct
     2701 cgacacagca ggtcaagagg agtacagtgc aatgagggac cagtacatga ggactgggga
     2761 gggctttctt tgtgtatttg ccataaataa tactaaatca tttgaagata ttcaccatta
     2821 tagagaacaa attaaaagag ttaaggactc tgaagatgta cctatggtcc tagtaggaaa
     2881 taaatgtgat ttgccttcta gaacagtaga cacaaaacag gctcaggact tagcaagaag
     2941 ttatggaatt ccttttattg aaacatcagc aaagacaaga cagggtgttg atgatgcctt
     3001 ctatacatta gttcgagaaa ttcgaaaaca taaagaaaag atgagcaaag atggtaaaaa
     3061 gaagaaaaag aagtcaaaga caaagtgtgt aattatgtga ttgccaactt tcttgtacaa
     3121 agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     3181 tcaaaataaa atcattattt gccatccagc tgatatcccc tatagtgagt cgtattacat
     3241 ggtcatagct gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc
     3301 acaagataaa aatatatcat catgcctcct ctagaccagc caggacagaa atgcctcgac
//
