LOCUS       sequence-164654-        4404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing GRB10 , part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4404)
  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 4404)
  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_GRB10_WT
FEATURES             Location/Qualifiers
     source          1..4404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2283..2382)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2399..2418)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2400..2418)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2423..2439)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2423..2439)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2870..3661
                     /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      3754..4342
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4243..4262
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agttggcacc atgaatgcat ccctggagag cctgtactcg gcctgcagca tgcagtcaga
      721 cacggtgccc ctcctgcaga atggccagca tgcccgcagc cagcctcggg cttcaggccc
      781 tcctcggtcc atccagccac aggtgtcccc aaggcagagg gtgcagcgct cccagcctgt
      841 gcacatcctc gctgtcaggc gccttcagga ggaagaccag cagtttagaa cctcatctct
      901 gccggccatc cccaatcctt ttcctgaact ctgtggccct gggagccccc ctgtgctcac
      961 gccgggttct ttacctccga gccaggccgc cgcaaagcag gatgttaaag tctttagtga
     1021 agatgggaca agcaaagtgg tggagattct agcagacatg acagccagag acctgtgcca
     1081 attgctggtt tacaaaagtc actgtgtgga tgacaacagc tggacactag tggagcacca
     1141 cccgcaccta ggattagaga ggtgcttgga agaccatgag ctggtggtcc aggtggagag
     1201 taccatggcc agtgagagta aatttctatt caggaagaat tacgcaaaat acgagttctt
     1261 taaaaatccc atgaatttct tcccagaaca gatggttact tggtgccagc agtcaaatgg
     1321 cagtcaaacc cagcttttgc agaattttct gaactccagt agttgtcctg aaattcaagg
     1381 gtttttgcat gtgaaagagc tgggaaagaa atcatggaaa aagctgtatg tgtgtttgcg
     1441 gagatctggc ctttattgct ccaccaaggg aacttcaaag gaacccagac acctgcagct
     1501 gctggccgac ctggaggaca gcaacatctt ctccctgatc gctggcagga agcagtacaa
     1561 cgcccctaca gaccacgggc tctgcataaa gccaaacaaa gtcaggaatg aaactaaaga
     1621 gctgaggttg ctctgtgcag aggacgagca aaccaggacg tgctggatga cagcgttcag
     1681 actcctcaag tatggaatgc tcctttacca gaattaccga atccctcagc agaggaaggc
     1741 cttgctgtcc ccgttctcga cgccagtgcg cagtgtctcc gagaactccc tcgtggcaat
     1801 ggatttttct gggcaaacag gacgcgtgat agagaatccg gcggaggccc agagcgcagc
     1861 cctggaggag ggccacgcct ggaggaagcg aagcacacgg atgaacatcc taggtagcca
     1921 aagtcccctc cacccttcta ccctaagtac agtgattcac aggacacagc actggtttca
     1981 cgggaggatc tccagggagg aatcccacag gatcattaaa cagcaagggc tcgtggatgg
     2041 gctttttctc ctccgtgaca gccagagtaa tccaaaggca tttgtactca cactgtgtca
     2101 tcaccagaaa attaaaaatt tccagatctt accttgcgag gacgacgggc agacgttctt
     2161 cagcctagat gacgggaaca ccaaattctc tgacctgatc cagctggttg acttttacca
     2221 gctgaacaaa ggagtcctgc cttgcaaact caagcaccac tgcatccgag tggccttatg
     2281 attgccaact ttcttgtaca aagttggcat tataagaaag cattgcttat caatttgttg
     2341 caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag ctgatatccc
     2401 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca
     2461 aaatctctga tgttacattg cacaagataa aaatatatca tcatgcctcc tctagaccag
     2521 ccaggacaga aatgcctcga cttcgctgct acccaaggtt gccgggtgac gcacaccgtg
     2581 gaaacggatg aaggcacgaa cccagtggac ataagcctgt tcggttcgta agctgtaatg
     2641 caagtagcgt atgcgctcac gcaactggtc cagaaccttg accgaacgca gcggtggtaa
     2701 cggcgcagtg gcggttttca tggcttgtta tgactgtttt tttggggtac agtctatgcc
     2761 tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag
     2821 caacgatgtt acgcagcagg gcagtcgccc taaaacaaag ttaaacatta tgagggaagc
     2881 ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg agcgccatct
     2941 cgaaccgacg ttgctggccg tacatttgta cggctccgca gtggatggcg gcctgaagcc
     3001 acacagtgat attgatttgc tggttacggt gaccgtaagg cttgatgaaa caacgcggcg
     3061 agctttgatc aacgaccttt tggaaacttc ggcttcccct ggagagagcg agattctccg
     3121 cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa
     3181 gcgcgaactg caatttggag aatggcagcg caatgacatt cttgcaggta tcttcgagcc
     3241 agccacgatc gacattgatc tggctatctt gctgacaaaa gcaagagaac atagcgttgc
     3301 cttggtaggt ccagcggcgg aggaactctt tgatccggtt cctgaacagg atctatttga
     3361 ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc gactgggctg gcgatgagcg
     3421 aaatgtagtg cttacgttgt cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc
     3481 gaaggatgtc gctgccgact gggcaatgga gcgcctgccg gcccagtatc agcccgtcat
     3541 acttgaagct agacaggctt atcttggaca agaagaagat cgcttggcct cgcgcgcaga
     3601 tcagttggaa gaatttgtcc actacgtgaa aggcgagatc accaaggtag tcggcaaata
     3661 accctcgagc cacccatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     3721 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3781 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3841 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3901 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3961 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4021 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4081 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4141 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4201 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4261 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4321 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4381 ttgctggcct tttgctcaca tgtt
//
