LOCUS       sequence-166769-        4716 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 4716)
  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 4716)
  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.H366Q
FEATURES             Location/Qualifiers
     source          1..4716
                     /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(2116..2215)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2232..2251)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2233..2251)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2256..2272)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2256..2272)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2703..3494
                     /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      3587..4175
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4076..4095
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4329..4346
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4505..4532)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4624..4710)
                     /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 ttcgaaaaga 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 cctcttcagg 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 aaaattgcca actttcttgt acaaagttgg cattataaga aagcattgct
     2161 tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc
     2221 cagctgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct
     2281 ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgcc
     2341 tcctctagac cagccaggac agaaatgcct cgacttcgct gctacccaag gttgccgggt
     2401 gacgcacacc gtggaaacgg atgaaggcac gaacccagtg gacataagcc tgttcggttc
     2461 gtaagctgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac
     2521 gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg
     2581 tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga
     2641 tgttatggag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca
     2701 ttatgaggga agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca
     2761 tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg
     2821 gcggcctgaa gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg
     2881 aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga
     2941 gcgagattct ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc
     3001 gttatccagc taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag
     3061 gtatcttcga gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag
     3121 aacatagcgt tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac
     3181 aggatctatt tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg
     3241 ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg
     3301 gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt
     3361 atcagcccgt catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg
     3421 cctcgcgcgc agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg
     3481 tagtcggcaa ataaccctcg agccacccat gaccaaaatc ccttaacgtg agttacgcgt
     3541 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3601 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3661 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3721 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3781 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3841 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3901 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3961 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4021 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4081 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4141 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4201 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     4261 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     4321 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     4381 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     4441 cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa
     4501 cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg
     4561 cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg
     4621 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
     4681 gtcttccgac tgagcctttc gttttatttg atgcct
//
