LOCUS       sequence-162653-        3756 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing RBKS, used as an over expression 
            control for gene expression analysis. Part of the Target Accelerator
            Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3756)
  AUTHORS   Berger AH, Brooks AN, Wu X, Shrestha Y, Chouinard C, Piccioni F, 
            Bagul M, Kamburov A, Imielinski M, Hogstrom L, Zhu C, Yang X, Pantel
            S, Sakai R, Watson J, Kaplan N, Campbell JD, Singh S, Root DE, 
            Narayan R, Natoli T, Lahr DL, Tirosh I, Tamayo P, Getz G, Wong B, 
            Doench J, Subramanian A, Golub TR, Meyerson M, Boehm JS
  TITLE     High-throughput Phenotyping of Lung Cancer Somatic Mutations.
  JOURNAL   Cancer Cell. 2016 Aug 8;30(2):214-28. doi: 
            10.1016/j.ccell.2016.06.022. Epub 2016 Jul 28.
  PUBMED    27478040
REFERENCE   2  (bases 1 to 3756)
  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: 
            "10.1016/j.ccell.2016.06.022"; journalName: "Cancer Cell"; date: 
            "2016-08-8- 8"; volume: "30"; issue: "2"; pages: "214-28"
COMMENT     Sequence Label: pDONR223_RBKS_WT_V5
FEATURES             Location/Qualifiers
     source          1..3756
                     /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(1156..1255)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1272..1291)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1273..1291)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1296..1312)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1296..1312)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1743..2534
                     /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      2627..3215
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3116..3135
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3369..3386
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3545..3572)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3664..3750)
                     /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 aagttggcat ggcggcgtct ggggaacccc agaggcagtg gcaagaggag gtggcggcgg
      241 tggtagtggt gggctcctgc atgaccgacc tggtcagtct tacttctcgt ttgccaaaaa
      301 ctggagaaac catccatgga cataagtttt ttattggctt tggagggaaa ggtgccaacc
      361 agtgtgtcca agctgctcgg cttggagcaa tgacgtccat ggtgtgtaag gttggcaaag
      421 attcttttgg caatgattat atagaaaact taaaacagaa tgatatttct acagaattta
      481 catatcagac taaagatgct gctacaggaa ctgcttctat aattgtcaat aatgaaggcc
      541 agaatatcat tgtcatagtg gctggagcaa atttactttt gaatacggag gatctgaggg
      601 cagcagccaa tgtcattagc agagccaaag tcatggtctg ccagctcgaa ataactccag
      661 caacttcttt ggaagcccta acaatggccc gcaggagtgg agtgaaaacc ttgttcaatc
      721 cagcccctgc cattgctgac ctggatcccc agttctacac cctctcagat gtgttctgct
      781 gcaatgaaag tgaggctgag attttaactg gcctcacggt gggcagcgct gcagatgctg
      841 gggaggctgc attagtgctc ttgaaaaggg gctgccaggt ggtaatcatt accttagggg
      901 ctgaaggatg tgtggtgctg tcacagacag aacctgagcc aaagcacatt cccacagaga
      961 aagtcaaggc tgtggatacc acgggtgctg gtgacagctt tgtgggagct ctggccttct
     1021 acctggctta ctatccaaat ctgtccttgg aagacatgct caacagatcc aatttcattg
     1081 cagcagtcag tgtccaggct gcaggaacac agtcatctta cccttacaaa aaagaccttc
     1141 cgcttactct gttttgccca actttcttgt acaaagttgg cattataaga aagcattgct
     1201 tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc
     1261 cagctgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct
     1321 ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgcc
     1381 tcctctagac cagccaggac agaaatgcct cgacttcgct gctacccaag gttgccgggt
     1441 gacgcacacc gtggaaacgg atgaaggcac gaacccagtg gacataagcc tgttcggttc
     1501 gtaagctgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac
     1561 gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg
     1621 tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga
     1681 tgttatggag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca
     1741 ttatgaggga agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca
     1801 tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg
     1861 gcggcctgaa gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg
     1921 aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga
     1981 gcgagattct ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc
     2041 gttatccagc taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag
     2101 gtatcttcga gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag
     2161 aacatagcgt tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac
     2221 aggatctatt tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg
     2281 ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg
     2341 gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt
     2401 atcagcccgt catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg
     2461 cctcgcgcgc agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg
     2521 tagtcggcaa ataaccctcg agccacccat gaccaaaatc ccttaacgtg agttacgcgt
     2581 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     2641 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     2701 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     2761 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     2821 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     2881 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     2941 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3001 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3061 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     3121 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     3181 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3241 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     3301 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     3361 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     3421 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     3481 cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa
     3541 cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg
     3601 cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg
     3661 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
     3721 gtcttccgac tgagcctttc gttttatttg atgcct
//
