LOCUS       sequence-168667-        4578 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing EYS , part of the Target Accelerator
            Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4578)
  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 4578)
  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_EYS_p.H466N
FEATURES             Location/Qualifiers
     source          1..4578
                     /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(2457..2556)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2573..2592)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2574..2592)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2597..2613)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2597..2613)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3044..3835
                     /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      3928..4516
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4417..4436
                     /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 atgactgaca aatcaatcgt cattctgagc ctgatggttt ttcacagctc
      721 tttcataaat ggaaaaacat gtagacggca attggtggaa gaatggcatc cacaaccctc
      781 atcatatgtg gtaaattgga cactaacaga aaacatctgc ttggacttct acagagattg
      841 ctggtttttg ggtgtaaaca ctaaaataga tacttcaggc aatcaagctg ttccccagat
      901 ttgccctttg caaattcaat taggagatat ccttgtaatt tcttctgaac catctcttca
      961 atttccagaa ataaatttga tgaatgtttc tgaaacatct ttcgttggct gtgtgcaaaa
     1021 taccacaacg gaagatcagt tactttttgg ctgcagacta aaaggaatgc acactgttaa
     1081 ttctaagtgg ctgagtgttg ggacacatta ttttatcaca gttatggcaa gtggtccatc
     1141 accttgtcca ctgggacttc gactaaatgt gacagtgaaa cagcagttct gccaggaatc
     1201 tctgagttca gaattttgct ctggtcatgg taaatgtctt agtgaagctt ggagcaagac
     1261 atatagctgc cattgccagc ctccattttc tggaaaatac tgccaggaac ttgatgcatg
     1321 ttcttttaaa ccatgtaaaa ataatggcag ttgcattaat aaaagagaaa attgggatga
     1381 gcaagcatat gaatgtgtct gtcacccacc atttacagga aagaattgct cagaaataat
     1441 tggccagtgt caaccacatg tctgtttcca tggaaactgc agcaatatta cttcaaatag
     1501 tttcatttgt gaatgtgatg agcaattttc aggtccattc tgtgaggtgt cagcaaaacc
     1561 ttgtgtttct ctgctttttt ggaaaagagg aatttgccca aatagcagtt ctgcttatac
     1621 ttatgaatgc ccaaaaggat cttccagcca aaatggtgaa actgatgtca gtgagttttc
     1681 attagtacca tgtcagaatg gtactgactg catcaaaata tcaaatgatg ttatgtgcat
     1741 ctgttcacca atatttacag atttgctttg taagagcatt caaacatcat gtgagtcatt
     1801 tcctttgagg aataatgcta catgtaagaa atgtgagaaa gattatcctt gcagctgtat
     1861 atcaggattt actgaaaaaa actgtgagaa agcaattgac cactgtaaac tgctcagcat
     1921 caactgtctg aatgaagaat ggtgtttcaa tataattgga agattcaaat atgtatgcat
     1981 tccagggtgc acaaaaaatc catgttggtt tttgaagaat gtttacctaa ttcatcaaca
     2041 cctctgctac tgtggagtca ccttcaatgg tatttgccaa gataaaggtc ctgctcaatt
     2101 tgaatatgtg tggcaattgg gatttgcagg atctgaaggc gaaaagtgcc aaggggttat
     2161 tgatgcctat ttctttctgg ctgcaaactg cactgaagat gcaacctatg tgaacgatcc
     2221 tgaagataat aattcttcat gttggttccc acatgaaggc acaaaagaga tttgtgcaaa
     2281 tggatgcagt tgtttgagtg aagaagacag tcaggaatat cggtatctat gttttctcag
     2341 atgggctggc aacatgtatc tggaaaatac aactgatgat caagaaaatg agtgtcaaca
     2401 tgaagctgtt tgtaaagatg aaattaatag acccaggtat ttatgtatca tttgattgcc
     2461 aactttcttg tacaaagttg tcattataag aaagcattgc ttatcaattt gttgcaacga
     2521 acaggtcact atcagtcaaa ataaaatcat tatttgccat ccagctgata tcccctatag
     2581 tgagtcgtat tacatggtca tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct
     2641 ctgatgttac attgcacaag ataaaaatat atcatcatgc ctcctctaga ccagccagga
     2701 cagaaatgcc tcgacttcgc tgctacccaa ggttgccggg tgacgcacac cgtggaaacg
     2761 gatgaaggca cgaacccagt ggacataagc ctgttcggtt cgtaagctgt aatgcaagta
     2821 gcgtatgcgc tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc
     2881 agtggcggtt ttcatggctt gttatgactg tttttttggg gtacagtcta tgcctcgggc
     2941 atccaagcag caagcgcgtt acgccgtggg tcgatgtttg atgttatgga gcagcaacga
     3001 tgttacgcag cagggcagtc gccctaaaac aaagttaaac attatgaggg aagcggtgat
     3061 cgccgaagta tcgactcaac tatcagaggt agttggcgtc atcgagcgcc atctcgaacc
     3121 gacgttgctg gccgtacatt tgtacggctc cgcagtggat ggcggcctga agccacacag
     3181 tgatattgat ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt
     3241 gatcaacgac cttttggaaa cttcggcttc ccctggagag agcgagattc tccgcgctgt
     3301 agaagtcacc attgttgtgc acgacgacat cattccgtgg cgttatccag ctaagcgcga
     3361 actgcaattt ggagaatggc agcgcaatga cattcttgca ggtatcttcg agccagccac
     3421 gatcgacatt gatctggcta tcttgctgac aaaagcaaga gaacatagcg ttgccttggt
     3481 aggtccagcg gcggaggaac tctttgatcc ggttcctgaa caggatctat ttgaggcgct
     3541 aaatgaaacc ttaacgctat ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt
     3601 agtgcttacg ttgtcccgca tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga
     3661 tgtcgctgcc gactgggcaa tggagcgcct gccggcccag tatcagcccg tcatacttga
     3721 agctagacag gcttatcttg gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt
     3781 ggaagaattt gtccactacg tgaaaggcga gatcaccaag gtagtcggca aataaccctc
     3841 gagccaccca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga
     3901 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3961 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4021 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     4081 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4141 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4201 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4261 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4321 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4381 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4441 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4501 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4561 gccttttgct cacatgtt
//
