LOCUS       sequence-166742-        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.D240N
FEATURES             Location/Qualifiers
     source          1..4719
                     /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(2119..2218)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2235..2254)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2236..2254)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2259..2275)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2259..2275)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2706..3497
                     /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      3590..4178
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4079..4098
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4332..4349
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4508..4535)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4627..4713)
                     /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 aagttggcac catggcatta tcactggaag aattcgtcca ctcccttgac ctcaggaccc
      241 tacccagggt tctagaaatc caggcaggca tctatcttga aggctctatt tatgaaatgt
      301 ttggaaatga atgctgtttt tcaacaggag aagtgattaa aattactggt ctcaaagtta
      361 agaagatcat agctgaaatt tgtgagcaga ttgaaggttg tgagtctcta cagccatttg
      421 aactgcctat gaattttcca ggtcttttta agattgtggc tgataaaact ccatacctta
      481 ctatggaaga aatcacaagg accattcata ttggaccaag tagactaggg catccttgct
      541 tctatcatca gaaggatata aaactagaga acctcatcat aaagcagggt gagcaaatca
      601 tgctcaactc agttgaagag attgatggag aaataatggt gagctgtgca gtagcaagga
      661 atcatcaaac tcactcattt aatttgcctt tgtcacaaga aggagaattc tacgagtgtg
      721 aagatgaacg tatttacact ctaaaggaga ttgttgaatg gaagattcct aagaacagaa
      781 caagaactgt aaaccttaca gatttttcaa ataagtggga ctcaacgaat ccatttccta
      841 aagactttta tggtaccctg attctcaagc ctgtttatga aattcaaggt gtgatgaaat
      901 ttcgaaaaaa tataatccgc atcctcccca gtctagatgt cgaagtcaaa gacatcactg
      961 attcttacga tgctaactgg tttcttcagc tgttatcaac agaagatctt tttgaaatga
     1021 ctagtaaaga gttccccata gtgactgaag tcatagaagc acctgaagga aaccacctgc
     1081 cccaaagcat tttacagcct gggaaaacca ttgtgatcca caaaaagtac caggcatcaa
     1141 gaatcttagc ttcagaaatt agaagcaatt ttcctaaaag acacttcttg atccccacta
     1201 gctataaagg caagttcaag cggcgaccga gggagttccc aacggcctat gacctagaga
     1261 tcgctaagag tgaaaaggag cctcttcacg tggtggccac caaagcgttt cattcccctc
     1321 atgacaagct gtcatccgta tctgttgggg accagtttct ggtgcatcag tcagagacga
     1381 ctgaagtcct ctgtgaggga ataaaaaaag tggtgaatgt tctggcctgt gaaaaaatcc
     1441 tcaaaaagtc ctatgaggct gcgctgctcc ctttgtacat ggaaggaggt tttgtagagg
     1501 tgattcatga taagaaacag tacccgattt ctgagctctg taaacagttc cgtttgccct
     1561 tcaatgtgaa ggtgtctgtc agggatcttt ccattgaaga ggacgtgttg gctgccacac
     1621 caggactgca gttggaggag gacattacag actcttacct actcataagt gactttgcca
     1681 accccacgga gtgctgggaa attcctgtgg gccgcttgaa tatgactgtt cagttagtta
     1741 gtaatttctc tagggatgca gaaccatttc tagtcaggac tctggtagaa gagatcactg
     1801 aagagcaata ttacatgatg cggagatatg aaagctcagc ctcacatccc ccacctcgcc
     1861 ctccgaaaca cccctcagta gaggaaacaa agttaaccct gctaacctta gcagaagaaa
     1921 ggacggtaga cctgcccaag tctcccaagc gtcatcacgt agacataacc aagaaacttc
     1981 acccaaatca agctggcctg gattcaaaag tactgattgg tagtcagaat gatttggtgg
     2041 atgaagagaa agaaaggagc aaccgtgggg ccacagcagt agcagaaaca ttcaaaaatg
     2101 aaaaacatca aaaatgattg ccaactttct tgtacaaagt tggcattata agaaagcatt
     2161 gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc
     2221 atccagctga tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc
     2281 tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat
     2341 gcctcctcta gaccagccag gacagaaatg cctcgacttc gctgctaccc aaggttgccg
     2401 ggtgacgcac accgtggaaa cggatgaagg cacgaaccca gtggacataa gcctgttcgg
     2461 ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga accttgaccg
     2521 aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttg
     2581 gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt
     2641 tgatgttatg gagcagcaac gatgttacgc agcagggcag tcgccctaaa acaaagttaa
     2701 acattatgag ggaagcggtg atcgccgaag tatcgactca actatcagag gtagttggcg
     2761 tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg
     2821 atggcggcct gaagccacac agtgatattg atttgctggt tacggtgacc gtaaggcttg
     2881 atgaaacaac gcggcgagct ttgatcaacg accttttgga aacttcggct tcccctggag
     2941 agagcgagat tctccgcgct gtagaagtca ccattgttgt gcacgacgac atcattccgt
     3001 ggcgttatcc agctaagcgc gaactgcaat ttggagaatg gcagcgcaat gacattcttg
     3061 caggtatctt cgagccagcc acgatcgaca ttgatctggc tatcttgctg acaaaagcaa
     3121 gagaacatag cgttgccttg gtaggtccag cggcggagga actctttgat ccggttcctg
     3181 aacaggatct atttgaggcg ctaaatgaaa ccttaacgct atggaactcg ccgcccgact
     3241 gggctggcga tgagcgaaat gtagtgctta cgttgtcccg catttggtac agcgcagtaa
     3301 ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc ctgccggccc
     3361 agtatcagcc cgtcatactt gaagctagac aggcttatct tggacaagaa gaagatcgct
     3421 tggcctcgcg cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc gagatcacca
     3481 aggtagtcgg caaataaccc tcgagccacc catgaccaaa atcccttaac gtgagttacg
     3541 cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3601 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3661 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3721 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3781 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3841 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3901 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3961 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     4021 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     4081 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     4141 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     4201 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     4261 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     4321 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     4381 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     4441 aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa gagtttgtag
     4501 aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta
     4561 tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca aatccgctcc
     4621 cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc
     4681 ccagtcttcc gactgagcct ttcgttttat ttgatgcct
//
