LOCUS       sequence-166775-        5580 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing RB1 , part of the Target Accelerator
            Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5580)
  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 5580)
  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_RB1_p.V654L
FEATURES             Location/Qualifiers
     source          1..5580
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     43..65
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     57..74
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     58..74
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    90..189
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2980..3079)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3096..3115)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3097..3115)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3120..3136)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3120..3136)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3567..4358
                     /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      4451..5039
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4940..4959
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5193..5210
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5369..5396)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5488..5574)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta
       61 aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga
      121 cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa
      181 aagttggcac catgccgccc aaaacccccc gaaaaacggc cgccaccgcc gccgctgccg
      241 ccgcggaacc cccggcaccg ccgccgccgc cccctcctga ggaggaccca gagcaggaca
      301 gcggcccgga ggacctgcct ctcgtcaggc ttgagtttga agaaacagaa gaacctgatt
      361 ttactgcatt atgtcagaaa ttaaagatac cagatcatgt cagagagaga gcttggttaa
      421 cttgggagaa agtttcatct gtggatggag tattgggagg ttatattcaa aagaaaaagg
      481 aactgtgggg aatctgtatc tttattgcag cagttgacct agatgagatg tcgttcactt
      541 ttactgagct acagaaaaac atagaaatca gtgtccataa attctttaac ttactaaaag
      601 aaattgatac cagtaccaaa gttgataatg ctatgtcaag actgttgaag aagtatgatg
      661 tattgtttgc actcttcagc aaattggaaa ggacatgtga acttatatat ttgacacaac
      721 ccagcagttc gatatctact gaaataaatt ctgcattggt gctaaaagtt tcttggatca
      781 catttttatt agctaaaggg gaagtattac aaatggaaga tgatctggtg atttcatttc
      841 agttaatgct atgtgtcctt gactatttta ttaaactctc acctcccatg ttgctcaaag
      901 aaccatataa aacagctgtt atacccatta atggttcacc tcgaacaccc aggcgaggtc
      961 agaacaggag tgcacggata gcaaaacaac tagaaaatga tacaagaatt attgaagttc
     1021 tctgtaaaga acatgaatgt aatatagatg aggtgaaaaa tgtttatttc aaaaatttta
     1081 taccttttat gaattctctt ggacttgtaa catctaatgg acttccagag gttgaaaatc
     1141 tttctaaacg atacgaagaa atttatctta aaaataaaga tctagatgca agattatttt
     1201 tggatcatga taaaactctt cagactgatt ctatagacag ttttgaaaca cagagaacac
     1261 cacgaaaaag taaccttgat gaagaggtga atgtaattcc tccacacact ccagttagga
     1321 ctgttatgaa cactatccaa caattaatga tgattttaaa ttcagcaagt gatcaacctt
     1381 cagaaaatct gatttcctat tttaacaact gcacagtgaa tccaaaagaa agtatactga
     1441 aaagagtgaa ggatatagga tacatcttta aagagaaatt tgctaaagct gtgggacagg
     1501 gttgtgtcga aattggatca cagcgataca aacttggagt tcgcttgtat taccgagtaa
     1561 tggaatccat gcttaaatca gaagaagaac gattatccat tcaaaatttt agcaaacttc
     1621 tgaatgacaa catttttcat atgtctttat tggcgtgcgc tcttgaggtt gtaatggcca
     1681 catatagcag aagtacatct cagaatcttg attctggaac agatttgtct ttcccatgga
     1741 ttctgaatgt gcttaattta aaagcctttg atttttacaa agtgatcgaa agttttatca
     1801 aagcagaagg caacttgaca agagaaatga taaaacattt agaacgatgt gaacatcgaa
     1861 tcatggaatc ccttgcatgg ctctcagatt cacctttatt tgatcttatt aaacaatcaa
     1921 aggaccgaga aggaccaact gatcaccttg aatctgcttg tcctcttaat cttcctctcc
     1981 agaataatca cactgcagca gatatgtatc tttctcctgt aagatctcca aagaaaaaag
     2041 gttcaactac gcgtgtaaat tctactgcaa atgcagagac acaagcaacc tcagccttcc
     2101 agacccagaa gccattgaaa tctacctctc tttcactgtt ttataaaaaa ctgtatcggc
     2161 tagcctatct ccggctaaat acactttgtg aacgccttct gtctgagcac ccagaattag
     2221 aacatatcat ctggaccctt ttccagcaca ccctgcagaa tgagtatgaa ctcatgagag
     2281 acaggcattt ggaccaaatt atgatgtgtt ccatgtatgg catatgcaaa gtgaagaata
     2341 tagaccttaa attcaaaatc attgtaacag catacaagga tcttcctcat gctgttcagg
     2401 agacattcaa acgtgttttg atcaaagaag aggagtatga ttctattata gtattctata
     2461 actcggtctt catgcagaga ctgaaaacaa atattttgca gtatgcttcc accaggcccc
     2521 ctaccttgtc accaatacct cacattcctc gaagccctta caagtttcct agttcaccct
     2581 tacggattcc tggagggaac atctatattt cacccctgaa gagtccatat aaaatttcag
     2641 aaggtctgcc aacaccaaca aaaatgactc caagatcaag aatcttagta tcaattggtg
     2701 aatcattcgg gacttctgag aagttccaga aaataaatca gatggtatgt aacagcgacc
     2761 gtgtgctcaa aagaagtgct gaaggaagca accctcctaa accactgaaa aaactacgct
     2821 ttgatattga aggatcagat gaagcagatg gaagtaaaca tctcccagga gagtccaaat
     2881 ttcagcagaa actggcagaa atgacttcta ctcgaacacg aatgcaaaag cagaaaatga
     2941 atgatagcat ggatacctca aacaaggaag agaaatgatt gccaactttc ttgtacaaag
     3001 ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc
     3061 aaaataaaat cattatttgc catccagctg atatccccta tagtgagtcg tattacatgg
     3121 tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt tacattgcac
     3181 aagataaaaa tatatcatca tgcctcctct agaccagcca ggacagaaat gcctcgactt
     3241 cgctgctacc caaggttgcc gggtgacgca caccgtggaa acggatgaag gcacgaaccc
     3301 agtggacata agcctgttcg gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca
     3361 actggtccag aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg
     3421 cttgttatga ctgttttttt ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc
     3481 gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagggca
     3541 gtcgccctaa aacaaagtta aacattatga gggaagcggt gatcgccgaa gtatcgactc
     3601 aactatcaga ggtagttggc gtcatcgagc gccatctcga accgacgttg ctggccgtac
     3661 atttgtacgg ctccgcagtg gatggcggcc tgaagccaca cagtgatatt gatttgctgg
     3721 ttacggtgac cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg
     3781 aaacttcggc ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc accattgttg
     3841 tgcacgacga catcattccg tggcgttatc cagctaagcg cgaactgcaa tttggagaat
     3901 ggcagcgcaa tgacattctt gcaggtatct tcgagccagc cacgatcgac attgatctgg
     3961 ctatcttgct gacaaaagca agagaacata gcgttgcctt ggtaggtcca gcggcggagg
     4021 aactctttga tccggttcct gaacaggatc tatttgaggc gctaaatgaa accttaacgc
     4081 tatggaactc gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc
     4141 gcatttggta cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg
     4201 caatggagcg cctgccggcc cagtatcagc ccgtcatact tgaagctaga caggcttatc
     4261 ttggacaaga agaagatcgc ttggcctcgc gcgcagatca gttggaagaa tttgtccact
     4321 acgtgaaagg cgagatcacc aaggtagtcg gcaaataacc ctcgagccac ccatgaccaa
     4381 aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca
     4441 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4501 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4561 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     4621 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4681 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4741 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4801 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4861 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4921 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4981 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5041 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     5101 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     5161 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     5221 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     5281 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg
     5341 ctagccagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt
     5401 agtttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt
     5461 cacaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa
     5521 acaacagata aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct
//
