LOCUS       sequence-162399-        4230 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing CFLAR, 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 4230)
  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 4230)
  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_CFLAR_WT_V5
FEATURES             Location/Qualifiers
     source          1..4230
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(3..22)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4..22)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(27..43)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(27..43)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             474..1265
                     /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      1358..1946
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1847..1866
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2100..2117
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2276..2303)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2395..2481)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2530..2552
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2544..2561
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2545..2561
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2577..2676
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(4117..4216)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt
       61 ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatgc ctcctctaga
      121 ccagccagga cagaaatgcc tcgacttcgc tgctacccaa ggttgccggg tgacgcacac
      181 cgtggaaacg gatgaaggca cgaacccagt ggacataagc ctgttcggtt cgtaagctgt
      241 aatgcaagta gcgtatgcgc tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg
      301 gtaacggcgc agtggcggtt ttcatggctt gttatgactg tttttttggg gtacagtcta
      361 tgcctcgggc atccaagcag caagcgcgtt acgccgtggg tcgatgtttg atgttatgga
      421 gcagcaacga tgttacgcag cagggcagtc gccctaaaac aaagttaaac attatgaggg
      481 aagcggtgat cgccgaagta tcgactcaac tatcagaggt agttggcgtc atcgagcgcc
      541 atctcgaacc gacgttgctg gccgtacatt tgtacggctc cgcagtggat ggcggcctga
      601 agccacacag tgatattgat ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc
      661 ggcgagcttt gatcaacgac cttttggaaa cttcggcttc ccctggagag agcgagattc
      721 tccgcgctgt agaagtcacc attgttgtgc acgacgacat cattccgtgg cgttatccag
      781 ctaagcgcga actgcaattt ggagaatggc agcgcaatga cattcttgca ggtatcttcg
      841 agccagccac gatcgacatt gatctggcta tcttgctgac aaaagcaaga gaacatagcg
      901 ttgccttggt aggtccagcg gcggaggaac tctttgatcc ggttcctgaa caggatctat
      961 ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc gcccgactgg gctggcgatg
     1021 agcgaaatgt agtgcttacg ttgtcccgca tttggtacag cgcagtaacc ggcaaaatcg
     1081 cgccgaagga tgtcgctgcc gactgggcaa tggagcgcct gccggcccag tatcagcccg
     1141 tcatacttga agctagacag gcttatcttg gacaagaaga agatcgcttg gcctcgcgcg
     1201 cagatcagtt ggaagaattt gtccactacg tgaaaggcga gatcaccaag gtagtcggca
     1261 aataaccctc gagccaccca tgaccaaaat cccttaacgt gagttacgcg tcgttccact
     1321 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     1381 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1441 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     1501 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     1561 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     1621 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     1681 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     1741 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     1801 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     1861 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     1921 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     1981 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     2041 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     2101 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     2161 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     2221 agcgcaacgc aattaatacg cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag
     2281 gccatccgtc aggatggcct tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc
     2341 tgcccgccac cctccgggcc gttgcttcac aacgttcaaa tccgctcccg gcggatttgt
     2401 cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtcttccga
     2461 ctgagccttt cgttttattt gatgcctggc agttccctac tctcgcgtta acgctagcat
     2521 ggatgttttc ccagtcacga cgttgtaaaa cgacggccag tcttaagctc gggccccaaa
     2581 taatgatttt attttgactg atagtgacct gttcgttgca acaaattgat gagcaatgct
     2641 tttttataat gccaactttg tacaaaaaag ttggcatgtc tgctgaagtc atccatcagg
     2701 ttgaagaagc acttgataca gatgagaagg agatgctgct ctttttgtgc cgggatgttg
     2761 ctatagatgt ggttccacct aatgtcaggg accttctgga tattttacgg gaaagaggta
     2821 agctgtctgt cggggacttg gctgaactgc tctacagagt gaggcgattt gacctgctca
     2881 aacgtatctt gaagatggac agaaaagctg tggagaccca cctgctcagg aaccctcacc
     2941 ttgtttcgga ctatagagtg ctgatggcag agattggtga ggatttggat aaatctgatg
     3001 tgtcctcatt aattttcctc atgaaggatt acatgggccg aggcaagata agcaaggaga
     3061 agagtttctt ggaccttgtg gttgagttgg agaaactaaa tctggttgcc ccagatcaac
     3121 tggatttatt agaaaaatgc ctaaagaaca tccacagaat agacctgaag acaaaaatcc
     3181 agaagtacaa gcagtctgtt caaggagcag ggacaagtta caggaatgtt ctccaagcag
     3241 caatccaaaa gagtctcaag gatccttcaa ataacttcag gctccataat gggagaagta
     3301 aagaacaaag acttaaggaa cagcttggcg ctcaacaaga accagtgaag aaatccattc
     3361 aggaatcaga agcttttttg cctcagagca tacctgaaga gagatacaag atgaagagca
     3421 agcccctagg aatctgcctg ataatcgatt gcattggcaa tgagacagag cttcttcgag
     3481 acaccttcac ttccctgggc tatgaagtcc agaaattctt gcatctcagt atgcatggta
     3541 tatcccagat tcttggccaa tttgcctgta tgcccgagca ccgagactac gacagctttg
     3601 tgtgtgtcct ggtgagccga ggaggctccc agagtgtgta tggtgtggat cagactcact
     3661 cagggctccc cctgcatcac atcaggagga tgttcatggg agattcatgc ccttatctag
     3721 cagggaagcc aaagatgttt tttattcaga actatgtggt gtcagagggc cagctggagg
     3781 acagcagcct cttggaggtg gatgggccag cgatgaagaa tgtggaattc aaggctcaga
     3841 agcgagggct gtgcacagtt caccgagaag ctgacttctt ctggagcctg tgtactgcgg
     3901 acatgtccct gctggagcag tctcacagct caccatccct gtacctgcag tgcctctccc
     3961 agaaactgag acaagaaaga aaacgcccac tcctggatct tcacattgaa ctcaatggct
     4021 acatgtatga ttggaacagc agagtttctg ccaaggagaa atattatgtc tggctgcagc
     4081 acactctgag aaagaaactt atcctctcct acacataccc aactttcttg tacaaagttg
     4141 gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa
     4201 ataaaatcat tatttgccat ccagctgata
//
