LOCUS       sequence-166507-        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.D33E
FEATURES             Location/Qualifiers
     source          1..3360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             311..1102
                     /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      1195..1783
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1684..1703
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1937..1954
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2113..2140)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2232..2318)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2367..2389
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2381..2398
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2382..2398
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2414..2513
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2516..3082
                     /codon_start=1
                     /product="human small GTPase proto-oncoprotein"
                     /label=K-Ras
                     /translation="MTEYKLVVVGAGGVGKSALTIQLIQNHFVDEYEPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
                     DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
                     RKHKEKMSKDGKKKKKKSKTKCVIM"
     protein_bind    complement(3084..3183)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3200..3219)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3201..3219)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3224..3240)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3224..3240)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 tgccgggtga cgcacaccgt ggaaacggat gaaggcacga acccagtgga cataagcctg
       61 ttcggttcgt aagctgtaat gcaagtagcg tatgcgctca cgcaactggt ccagaacctt
      121 gaccgaacgc agcggtggta acggcgcagt ggcggttttc atggcttgtt atgactgttt
      181 ttttggggta cagtctatgc ctcgggcatc caagcagcaa gcgcgttacg ccgtgggtcg
      241 atgtttgatg ttatggagca gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa
      301 gttaaacatt atgagggaag cggtgatcgc cgaagtatcg actcaactat cagaggtagt
      361 tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc gtacatttgt acggctccgc
      421 agtggatggc ggcctgaagc cacacagtga tattgatttg ctggttacgg tgaccgtaag
      481 gcttgatgaa acaacgcggc gagctttgat caacgacctt ttggaaactt cggcttcccc
      541 tggagagagc gagattctcc gcgctgtaga agtcaccatt gttgtgcacg acgacatcat
      601 tccgtggcgt tatccagcta agcgcgaact gcaatttgga gaatggcagc gcaatgacat
      661 tcttgcaggt atcttcgagc cagccacgat cgacattgat ctggctatct tgctgacaaa
      721 agcaagagaa catagcgttg ccttggtagg tccagcggcg gaggaactct ttgatccggt
      781 tcctgaacag gatctatttg aggcgctaaa tgaaacctta acgctatgga actcgccgcc
      841 cgactgggct ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt ggtacagcgc
      901 agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg agcgcctgcc
      961 ggcccagtat cagcccgtca tacttgaagc tagacaggct tatcttggac aagaagaaga
     1021 tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc cactacgtga aaggcgagat
     1081 caccaaggta gtcggcaaat aaccctcgag ccacccatga ccaaaatccc ttaacgtgag
     1141 ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     1201 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     1261 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     1321 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     1381 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     1441 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     1501 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     1561 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     1621 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     1681 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     1741 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     1801 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     1861 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     1921 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     1981 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     2041 ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt accgctagcc aggaagagtt
     2101 tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttagtttg atgcctggca
     2161 gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc
     2221 gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga
     2281 aaggcccagt cttccgactg agcctttcgt tttatttgat gcctggcagt tccctactct
     2341 cgcgttaacg ctagcatgga tgttttccca gtcacgacgt tgtaaaacga cggccagtct
     2401 taagctcggg ccccaaataa tgattttatt ttgactgata gtgacctgtt cgttgcaaca
     2461 aattgatgag caatgctttt ttataatgcc aactttgtac aaaaaagttg gcaccatgac
     2521 tgaatataaa cttgtggtag ttggagctgg tggcgtaggc aagagtgcct tgacgataca
     2581 gctaattcag aatcattttg tggacgaata tgaaccaaca atagaggatt cctacaggaa
     2641 gcaagtagta attgatggag aaacctgtct cttggatatt ctcgacacag caggtcaaga
     2701 ggagtacagt gcaatgaggg accagtacat gaggactggg gagggctttc tttgtgtatt
     2761 tgccataaat aatactaaat catttgaaga tattcaccat tatagagaac aaattaaaag
     2821 agttaaggac tctgaagatg tacctatggt cctagtagga aataaatgtg atttgccttc
     2881 tagaacagta gacacaaaac aggctcagga cttagcaaga agttatggaa ttccttttat
     2941 tgaaacatca gcaaagacaa gacagggtgt tgatgatgcc ttctatacat tagttcgaga
     3001 aattcgaaaa cataaagaaa agatgagcaa agatggtaaa aagaagaaaa agaagtcaaa
     3061 gacaaagtgt gtaattatgt gattgccaac tttcttgtac aaagttggca ttataagaaa
     3121 gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat
     3181 ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     3241 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     3301 atcatgcctc ctctagacca gccaggacag aaatgcctcg acttcgctgc tacccaaggt
//
