LOCUS       sequence-163006-        3693 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing CGREF1, 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 3693)
  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 3693)
  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_CGREF1_WT_V5
FEATURES             Location/Qualifiers
     source          1..3693
                     /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(1093..1192)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1209..1228)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1210..1228)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1233..1249)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1233..1249)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1680..2471
                     /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      2564..3152
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3053..3072
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3306..3323
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3482..3509)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3601..3687)
                     /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 gttacctttg acgatgacag tgttaatcct gctgctgctc cccacgggtc
      241 aggctgcccc aaaggatgga gtcacaaggc cagactctga agtgcagcat cagctcctgc
      301 ccaacccctt ccagccaggc caggagcagc tcggacttct gcagagctac ctaaagggac
      361 taggaaggac agaagtgcaa ctggagcatc tgagccggga gcaggttctc ctctacctct
      421 ttgccctcca tgactatgac cagagtggac agctggatgg cctggagctg ctgtccatgt
      481 tgacagctgc tctggcccct ggagctgcca actctcctac caccaacccg gtgatattga
      541 tagtggacaa agtgctcgag acgcaggacc tgaatgggga tgggctcatg acccctgctg
      601 agctcatcaa cttcccggga gtagccctca ggcacgtgga gcccggagag ccccttgctc
      661 catctcctca ggagccacaa gctgttggaa ggcagtccct attagctaaa agcccattaa
      721 gacaagaaac acaggaagcc cctggtccca gagaagaagc aaagggccag gtagaggcca
      781 gaagggagtc tttggatcct gtccaggagc ctgggggcca ggcagaggct gatggagatg
      841 ttccagggcc cagaggggaa gctgagggcc aggcagaggc taaaggagat gcccctgggc
      901 ccagagggga agctgggggc caggcagagg ctgaaggaga tgcccccggg cccagagggg
      961 aagctggggg ccaggcagag gccagggaga atggagagga ggccaaggaa cttccagggg
     1021 aaacactgga gtctaagaac acccaaaatg actttgaggt gcacattgtt caagtggaga
     1081 atgatgagat ctrcccaact ttcttgtaca aagttggcat tataagaaag cattgcttat
     1141 caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag
     1201 ctgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc
     1261 ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca tcatgcctcc
     1321 tctagaccag ccaggacaga aatgcctcga cttcgctgct acccaaggtt gccgggtgac
     1381 gcacaccgtg gaaacggatg aaggcacgaa cccagtggac ataagcctgt tcggttcgta
     1441 agctgtaatg caagtagcgt atgcgctcac gcaactggtc cagaaccttg accgaacgca
     1501 gcggtggtaa cggcgcagtg gcggttttca tggcttgtta tgactgtttt tttggggtac
     1561 agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt
     1621 tatggagcag caacgatgtt acgcagcagg gcagtcgccc taaaacaaag ttaaacatta
     1681 tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg
     1741 agcgccatct cgaaccgacg ttgctggccg tacatttgta cggctccgca gtggatggcg
     1801 gcctgaagcc acacagtgat attgatttgc tggttacggt gaccgtaagg cttgatgaaa
     1861 caacgcggcg agctttgatc aacgaccttt tggaaacttc ggcttcccct ggagagagcg
     1921 agattctccg cgctgtagaa gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt
     1981 atccagctaa gcgcgaactg caatttggag aatggcagcg caatgacatt cttgcaggta
     2041 tcttcgagcc agccacgatc gacattgatc tggctatctt gctgacaaaa gcaagagaac
     2101 atagcgttgc cttggtaggt ccagcggcgg aggaactctt tgatccggtt cctgaacagg
     2161 atctatttga ggcgctaaat gaaaccttaa cgctatggaa ctcgccgccc gactgggctg
     2221 gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg gtacagcgca gtaaccggca
     2281 aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga gcgcctgccg gcccagtatc
     2341 agcccgtcat acttgaagct agacaggctt atcttggaca agaagaagat cgcttggcct
     2401 cgcgcgcaga tcagttggaa gaatttgtcc actacgtgaa aggcgagatc accaaggtag
     2461 tcggcaaata accctcgagc cacccatgac caaaatccct taacgtgagt tacgcgtcgt
     2521 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     2581 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     2641 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     2701 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     2761 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     2821 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     2881 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     2941 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3001 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3061 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3121 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3181 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     3241 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     3301 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     3361 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     3421 gcagtgagcg caacgcaatt aatacgcgta ccgctagcca ggaagagttt gtagaaacgc
     3481 aaaaaggcca tccgtcagga tggccttctg cttagtttga tgcctggcag tttatggcgg
     3541 gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg
     3601 atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc
     3661 ttccgactga gcctttcgtt ttatttgatg cct
//
