LOCUS       sequence-168663-        4719 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing THEMIS , part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4719)
  AUTHORS   Kim E, Ilic N, Shrestha Y, Zou L, Kamburov A, Zhu C, Yang X, Lubonja
            R, Tran N, Nguyen C, Lawrence MS, Piccioni F, Bagul M, Doench JG, 
            Chouinard CR, Wu X, Hogstrom L, Natoli T, Tamayo P, Horn H, Corsello
            SM, Lage K, Root DE, Subramanian A, Golub TR, Getz G, Boehm JS, Hahn
            WC
  TITLE     Systematic Functional Interrogation of Rare Cancer Variants 
            Identifies Oncogenic Alleles.
  JOURNAL   Cancer Discov. 2016 Jul;6(7):714-26. doi: 
            10.1158/2159-8290.CD-16-0160. Epub 2016 May 4.
  PUBMED    27147599
REFERENCE   2  (bases 1 to 4719)
  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: 
            "10.1158/2159-8290.CD-16-0160"; journalName: "Cancer Discov"; date: 
            "2016-07"; volume: "6"; issue: "7"; pages: "714-26"
COMMENT     Sequence Label: pDONR223_THEMIS_p.R196I
FEATURES             Location/Qualifiers
     source          1..4719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2598..2697)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2714..2733)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2715..2733)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2738..2754)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2738..2754)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3185..3976
                     /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      4069..4657
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4558..4577
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agttggcacc atggcattat cactggaaga attcgtccac tcccttgacc tcaggaccct
      721 acccagggtt ctagaaatcc aggcaggcat ctatcttgaa ggctctattt atgaaatgtt
      781 tggaaatgaa tgctgttttt caacaggaga agtgattaaa attactggtc tcaaagttaa
      841 gaagatcata gctgaaattt gtgagcagat tgaaggttgt gagtctctac agccatttga
      901 actgcctatg aattttccag gtctttttaa gattgtggct gataaaactc cataccttac
      961 tatggaagaa atcacaagga ccattcatat tggaccaagt agactagggc atccttgctt
     1021 ctatcatcag aaggatataa aactagagaa cctcatcata aagcagggtg agcaaatcat
     1081 gctcaactca gttgaagaga ttgatggaga aataatggtg agctgtgcag tagcaaggaa
     1141 tcatcaaact cactcattta atttgccttt gtcacaagaa ggagaattct acgagtgtga
     1201 agatgaacgt atttacactc taaaggagat tgttgaatgg aagattccta agaacataac
     1261 aagaactgta aaccttacag atttttcaaa taagtgggac tcaacgaatc catttcctaa
     1321 agacttttat ggtaccctga ttctcaagcc tgtttatgaa attcaaggtg tgatgaaatt
     1381 tcgaaaagat ataatccgca tcctccccag tctagatgtc gaagtcaaag acatcactga
     1441 ttcttacgat gctaactggt ttcttcagct gttatcaaca gaagatcttt ttgaaatgac
     1501 tagtaaagag ttccccatag tgactgaagt catagaagca cctgaaggaa accacctgcc
     1561 ccaaagcatt ttacagcctg ggaaaaccat tgtgatccac aaaaagtacc aggcatcaag
     1621 aatcttagct tcagaaatta gaagcaattt tcctaaaaga cacttcttga tccccactag
     1681 ctataaaggc aagttcaagc ggcgaccgag ggagttccca acggcctatg acctagagat
     1741 cgctaagagt gaaaaggagc ctcttcacgt ggtggccacc aaagcgtttc attcccctca
     1801 tgacaagctg tcatccgtat ctgttgggga ccagtttctg gtgcatcagt cagagacgac
     1861 tgaagtcctc tgtgagggaa taaaaaaagt ggtgaatgtt ctggcctgtg aaaaaatcct
     1921 caaaaagtcc tatgaggctg cgctgctccc tttgtacatg gaaggaggtt ttgtagaggt
     1981 gattcatgat aagaaacagt acccgatttc tgagctctgt aaacagttcc gtttgccctt
     2041 caatgtgaag gtgtctgtca gggatctttc cattgaagag gacgtgttgg ctgccacacc
     2101 aggactgcag ttggaggagg acattacaga ctcttaccta ctcataagtg actttgccaa
     2161 ccccacggag tgctgggaaa ttcctgtggg ccgcttgaat atgactgttc agttagttag
     2221 taatttctct agggatgcag aaccatttct agtcaggact ctggtagaag agatcactga
     2281 agagcaatat tacatgatgc ggagatatga aagctcagcc tcacatcccc cacctcgccc
     2341 tccgaaacac ccctcagtag aggaaacaaa gttaaccctg ctaaccttag cagaagaaag
     2401 gacggtagac ctgcccaagt ctcccaagcg tcatcacgta gacataacca agaaacttca
     2461 cccaaatcaa gctggcctgg attcaaaagt actgattggt agtcagaatg atttggtgga
     2521 tgaagagaaa gaaaggagca accgtggggc cacagcagta gcagaaacat tcaaaaatga
     2581 aaaacatcaa aaatgattgc caactttctt gtacaaagtt ggcattataa gaaagcattg
     2641 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     2701 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     2761 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     2821 cctcctctag accagccagg acagaaatgc ctcgacttcg ctgctaccca aggttgccgg
     2881 gtgacgcaca ccgtggaaac ggatgaaggc acgaacccag tggacataag cctgttcggt
     2941 tcgtaagctg taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga
     3001 acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgg
     3061 ggtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt
     3121 gatgttatgg agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttaaa
     3181 cattatgagg gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt
     3241 catcgagcgc catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga
     3301 tggcggcctg aagccacaca gtgatattga tttgctggtt acggtgaccg taaggcttga
     3361 tgaaacaacg cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga
     3421 gagcgagatt ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg
     3481 gcgttatcca gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc
     3541 aggtatcttc gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag
     3601 agaacatagc gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga
     3661 acaggatcta tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg
     3721 ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac
     3781 cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca
     3841 gtatcagccc gtcatacttg aagctagaca ggcttatctt ggacaagaag aagatcgctt
     3901 ggcctcgcgc gcagatcagt tggaagaatt tgtccactac gtgaaaggcg agatcaccaa
     3961 ggtagtcggc aaataaccct cgagccaccc atgaccaaaa tcccttaacg tgagttacgc
     4021 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4081 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4141 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4201 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4261 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4321 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4381 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4441 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4501 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4561 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4621 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4681 acggttcctg gccttttgct ggccttttgc tcacatgtt
//
