LOCUS       sequence-162761-        3798 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing DARC, 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 3798)
  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 3798)
  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_DARC_WT_V5
FEATURES             Location/Qualifiers
     source          1..3798
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     705..721
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     705..721
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     726..745
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        726..744
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    762..861
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1870..1969)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1985..2002)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1985..2001)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1994..2016)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      2065..2151
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2243..2270
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2429..2446)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2600..3188)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2680..2699)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3281..3798,1..274))
                     /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 caacaatggt gacttctaca gcgcggagaa tctcgctctc tccaggggaa gccgaagttt
       61 ccaaaaggtc gttgatcaaa gctcgccgcg ttgtttcatc aagccttacg gtcaccgtaa
      121 ccagcaaatc aatatcactg tgtggcttca ggccgccatc cactgcggag ccgtacaaat
      181 gtacggccag caacgtcggt tcgagatggc gctcgatgac gccaactacc tctgatagtt
      241 gagtcgatac ttcggcgatc accgcttccc tcataatgtt taactttgtt ttagggcgac
      301 tgccctgctg cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc acggcgtaac
      361 gcgcttgctg cttggatgcc cgaggcatag actgtacccc aaaaaaacag tcataacaag
      421 ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt
      481 tgcgtgagcg catacgctac ttgcattaca gcttacgaac cgaacaggct tatgtccact
      541 gggttcgtgc cttcatccgt ttccacggtg tgcgtcaccc ggcaaccttg ggtagcagcg
      601 aagtcgaggc atttctgtcc tggctggtct agaggaggca tgatgatata tttttatctt
      661 gtgcaatgta acatcagaga ttttgagaca cgggccagag ctgccaggaa acagctatga
      721 ccatgtaata cgactcacta taggggatat cagctggatg gcaaataatg attttatttt
      781 gactgatagt gacctgttcg ttgcaacaaa ttgataagca atgctttctt ataatgccaa
      841 ctttgtacaa gaaagttggg taggatttgc ttccaagggt gtccagatga gaaaaccatc
      901 cttcagggag gggcagagag ggcaagaggg tgcgggtggc ctggtggcag aatagggcga
      961 ggagcagggg cgtagccaca cagtgcaaaa ttgccagggc ttctgccagg ttcagcagca
     1021 ggtccagagc ctgctgggcc agacatgttg acaacagcaa cagcttggac ctcaccagga
     1081 aatccagtcc tagaaccacc ccatgaggcc accagaaaat aaaccaggcc cacaggatgt
     1141 tcatccaggg gcctggcccc atacccaatg ccttcttcag ccccttggct ccaaacaaac
     1201 ccaatggcaa caagacaaag atggcaagac aggctacagt gtgtgtggcc tgcaaagcct
     1261 tcagctccgt gctgtatatc agggtgcaga gtccaccaga agcaccactg gccagggtga
     1321 caggcagtgt cagtagggca gccactcccc aaattcccac agtgagcccc agggtgaggc
     1381 ctgggacctg gcctgcaccc agtctgtggc ccagggaggc atggcaccct agcagcaaag
     1441 cctgggcaaa ggctgagcca taccagacac agtagcccag gctacacagg gcagagctgc
     1501 gagtgctacc tagccctggg gccaagacgg gcaccacaat gctgaagagg gcactgccca
     1561 cagccagctg tgccaggaca ggccagccag ggcagagctg ccagcggaag agaggtctga
     1621 aaagcatgaa gaggacagtg ctgctagcta ggatacccag gacactggtg aggatgaaga
     1681 agggcagtgc agagtcatcc agcaggttac aggagtggca gggggcagct gcttccaggt
     1741 tggcaccata gtctccatct gggaaggaat cattcacacc ataggaagaa ttccatacat
     1801 cttcgaagtc cagctgactt gagttctcag ttgaggggga gagctccgcc ctgtgcagac
     1861 agttccccat gccaactttt ttgtacaaag ttggcattat aaaaaagcat tgctcatcaa
     1921 tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgg ggcccgagct
     1981 taagactggc cgtcgtttta caacgtcgtg actgggaaaa catccatgct agcgttaacg
     2041 cgagagtagg gaactgccag gcatcaaata aaacgaaagg ctcagtcgga agactgggcc
     2101 tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga
     2161 gcggatttga acgttgtgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa
     2221 actgccaggc atcaaactaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta
     2281 caaactcttc ctggctagcg gtacgcgtat taattgcgtt gcgctcactg cccgctttcc
     2341 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     2401 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2461 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2521 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2581 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2641 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2701 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2761 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2821 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2881 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2941 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3001 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3061 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3121 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3181 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgacgcgt
     3241 aactcacgtt aagggatttt ggtcatgggt ggctcgaggg ttatttgccg actaccttgg
     3301 tgatctcgcc tttcacgtag tggacaaatt cttccaactg atctgcgcgc gaggccaagc
     3361 gatcttcttc ttgtccaaga taagcctgtc tagcttcaag tatgacgggc tgatactggg
     3421 ccggcaggcg ctccattgcc cagtcggcag cgacatcctt cggcgcgatt ttgccggtta
     3481 ctgcgctgta ccaaatgcgg gacaacgtaa gcactacatt tcgctcatcg ccagcccagt
     3541 cgggcggcga gttccatagc gttaaggttt catttagcgc ctcaaataga tcctgttcag
     3601 gaaccggatc aaagagttcc tccgccgctg gacctaccaa ggcaacgcta tgttctcttg
     3661 cttttgtcag caagatagcc agatcaatgt cgatcgtggc tggctcgaag atacctgcaa
     3721 gaatgtcatt gcgctgccat tctccaaatt gcagttcgcg cttagctgga taacgccacg
     3781 gaatgatgtc gtcgtgca
//
