LOCUS       sequence-162737-        3786 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing CGRRF1, 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 3786)
  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 3786)
  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_CGRRF1_WT_V5
FEATURES             Location/Qualifiers
     source          1..3786
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     238..254
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     238..254
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     259..278
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        259..277
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    295..394
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    840..851
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     protein_bind    complement(1391..1490)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1506..1523)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1506..1522)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1515..1537)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      1586..1672
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1764..1791
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1950..1967)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2121..2709)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2201..2220)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2802..3593)
                     /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 gttctggacc agttgcgtga gcgcatacgc tacttgcatt acagcttacg aaccgaacag
       61 gcttatgtcc actgggttcg tgccttcatc cgtttccacg gtgtgcgtca cccggcaacc
      121 ttgggtagca gcgaagtcga ggcatttctg tcctggctgg tctagaggag gcatgatgat
      181 atatttttat cttgtgcaat gtaacatcag agattttgag acacgggcca gagctgccag
      241 gaaacagcta tgaccatgta atacgactca ctatagggga tatcagctgg atggcaaata
      301 atgattttat tttgactgat agtgacctgt tcgttgcaac aaattgataa gcaatgcttt
      361 cttataatgc caactttgta caagaaagtt gggcaaagag tcttcggttt gtctttatct
      421 tgctcttttt gactgcaaag tgcaaaagat tcctgaacaa actgcctgca cattgggcac
      481 tgctgaaaat acttcacaca gccatcacac aggcatgtgt gtctgcatgg taagagtacc
      541 cagttcacag tcccattctg gcaaacaaca cagtccttgc tgttctcttc cgatggctca
      601 acttcacttt cagagagtcc caccttttcc aacaaacttc tgtctgtgtt tttttcttct
      661 gaagaggaat tgttggaggg agtgaaatta ttatttgcag acatgaaaag ttgcttaaga
      721 tcatgaaatt gaccttgagc caagagtaaa tattgataca atattctgca ggatagttta
      781 taagtcctat caggaatatg aatcactgac accatggaaa taatatcata aatttcccgg
      841 tcatcctcat cagctaaggt caatagcgct accaatggat agcgagatct gggtactgta
      901 ccaaagtctt caattttagt atctcttggt aactggcaat atatttcttc tttgctatcc
      961 tttttaataa aatactgttc ctgatagaga tattcactat acagagcatc ttctaatgct
     1021 tggggagtgc ttattctgaa gcaataaaca tgcttctgca gagcttcata taatttttga
     1081 acattgcacc cccagtagca tgtaaggagg ctatcttcaa ggcaatctgt tgtcaaggtt
     1141 atgccagttg taattgaagc tgaagatgga ttagtgatct ctaagccaaa aggattcttg
     1201 acttgtctca tttgcttttt gaaaactctt gtgctgaact gggtctcttc tgaatttctc
     1261 agaattactg gaacatccca accaaaccat cccaatacca ggccggtggt cacgatgaag
     1321 caggtaaaga ccaccgcgat gtagaaaagc ggcgagtatt cataaagcgt taccagaaac
     1381 accgcagcca tgccaacttt tttgtacaaa gttggcatta taaaaaagca ttgctcatca
     1441 atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg gggcccgagc
     1501 ttaagactgg ccgtcgtttt acaacgtcgt gactgggaaa acatccatgc tagcgttaac
     1561 gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcgg aagactgggc
     1621 ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg
     1681 agcggatttg aacgttgtga agcaacggcc cggagggtgg cgggcaggac gcccgccata
     1741 aactgccagg catcaaacta agcagaaggc catcctgacg gatggccttt ttgcgtttct
     1801 acaaactctt cctggctagc ggtacgcgta ttaattgcgt tgcgctcact gcccgctttc
     1861 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     1921 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     1981 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2041 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2101 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2161 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2221 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2281 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2341 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2401 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2461 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2521 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     2581 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     2641 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     2701 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgacgcg
     2761 taactcacgt taagggattt tggtcatggg tggctcgagg gttatttgcc gactaccttg
     2821 gtgatctcgc ctttcacgta gtggacaaat tcttccaact gatctgcgcg cgaggccaag
     2881 cgatcttctt cttgtccaag ataagcctgt ctagcttcaa gtatgacggg ctgatactgg
     2941 gccggcaggc gctccattgc ccagtcggca gcgacatcct tcggcgcgat tttgccggtt
     3001 actgcgctgt accaaatgcg ggacaacgta agcactacat ttcgctcatc gccagcccag
     3061 tcgggcggcg agttccatag cgttaaggtt tcatttagcg cctcaaatag atcctgttca
     3121 ggaaccggat caaagagttc ctccgccgct ggacctacca aggcaacgct atgttctctt
     3181 gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa gatacctgca
     3241 agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg ataacgccac
     3301 ggaatgatgt cgtcgtgcac aacaatggtg acttctacag cgcggagaat ctcgctctct
     3361 ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt tgtttcatca
     3421 agccttacgg tcaccgtaac cagcaaatca atatcactgt gtggcttcag gccgccatcc
     3481 actgcggagc cgtacaaatg tacggccagc aacgtcggtt cgagatggcg ctcgatgacg
     3541 ccaactacct ctgatagttg agtcgatact tcggcgatca ccgcttccct cataatgttt
     3601 aactttgttt tagggcgact gccctgctgc gtaacatcgt tgctgctcca taacatcaaa
     3661 catcgaccca cggcgtaacg cgcttgctgc ttggatgccc gaggcataga ctgtacccca
     3721 aaaaaacagt cataacaagc catgaaaacc gccactgcgc cgttaccacc gctgcgttcg
     3781 gtcaag
//
