LOCUS       sequence-166548-        3363 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing NRAS, part of the Target Accelerator
            Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3363)
  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 3363)
  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_NRAS_p.Y64D
FEATURES             Location/Qualifiers
     source          1..3363
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      39..627
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     528..547
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     781..798
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(957..984)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1076..1162)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1211..1233
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1225..1242
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1226..1242
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1258..1357
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1931..2030)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2047..2066)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2048..2066)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2071..2087)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2071..2087)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2518..3309
                     /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 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
       61 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
      121 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
      181 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
      241 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
      301 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
      361 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
      421 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
      481 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
      541 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
      601 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
      661 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
      721 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
      781 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
      841 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
      901 gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa
      961 ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc
     1021 ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg
     1081 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg
     1141 actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgctagca
     1201 tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaagct cgggccccaa
     1261 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga tgagcaatgc
     1321 ttttttataa tgccaacttt gtacaaaaaa gttggcacca tgactgagta caaactggtg
     1381 gtggttggag caggtggtgt tgggaaaagc gcactgacaa tccagctaat ccagaaccac
     1441 tttgtagatg aatatgatcc caccatagag gattcttaca gaaaacaagt ggttatagat
     1501 ggtgaaacct gtttgttgga catactggat acagctggac aagaagagga cagtgccatg
     1561 agagaccaat acatgaggac aggcgaaggc ttcctctgtg tatttgccat caataatagc
     1621 aagtcatttg cggatattaa cctctacagg gagcagatta agcgagtaaa agactcggat
     1681 gatgtaccta tggtgctagt gggaaacaag tgtgatttgc caacaaggac agttgataca
     1741 aaacaagccc acgaactggc caagagttac gggattccat tcattgaaac ctcagccaag
     1801 accagacagg gtgttgaaga tgctttttac acactggtaa gagaaatacg ccagtaccga
     1861 atgaaaaaac tcaacagcag tgatgatggg actcagggtt gtatgggatt gccatgtgtg
     1921 gtgatgtaat tgccaacttt cttgtacaaa gttggcatta taagaaagca ttgcttatca
     1981 atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct
     2041 gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc
     2101 gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc atgcctcctc
     2161 tagaccagcc aggacagaaa tgcctcgact tcgctgctac ccaaggttgc cgggtgacgc
     2221 acaccgtgga aacggatgaa ggcacgaacc cagtggacat aagcctgttc ggttcgtaag
     2281 ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac cgaacgcagc
     2341 ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt tggggtacag
     2401 tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta
     2461 tggagcagca acgatgttac gcagcagggc agtcgcccta aaacaaagtt aaacattatg
     2521 agggaagcgg tgatcgccga agtatcgact caactatcag aggtagttgg cgtcatcgag
     2581 cgccatctcg aaccgacgtt gctggccgta catttgtacg gctccgcagt ggatggcggc
     2641 ctgaagccac acagtgatat tgatttgctg gttacggtga ccgtaaggct tgatgaaaca
     2701 acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg cttcccctgg agagagcgag
     2761 attctccgcg ctgtagaagt caccattgtt gtgcacgacg acatcattcc gtggcgttat
     2821 ccagctaagc gcgaactgca atttggagaa tggcagcgca atgacattct tgcaggtatc
     2881 ttcgagccag ccacgatcga cattgatctg gctatcttgc tgacaaaagc aagagaacat
     2941 agcgttgcct tggtaggtcc agcggcggag gaactctttg atccggttcc tgaacaggat
     3001 ctatttgagg cgctaaatga aaccttaacg ctatggaact cgccgcccga ctgggctggc
     3061 gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa
     3121 atcgcgccga aggatgtcgc tgccgactgg gcaatggagc gcctgccggc ccagtatcag
     3181 cccgtcatac ttgaagctag acaggcttat cttggacaag aagaagatcg cttggcctcg
     3241 cgcgcagatc agttggaaga atttgtccac tacgtgaaag gcgagatcac caaggtagtc
     3301 ggcaaataac cctcgagcca cccatgacca aaatccctta acgtgagtta cgcgtcgttc
     3361 cac
//
