LOCUS       sequence-167266-        4296 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing SPAG8 , part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4296)
  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 4296)
  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_SPAG8_p.P344L
FEATURES             Location/Qualifiers
     source          1..4296
                     /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(2175..2274)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2291..2310)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2292..2310)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2315..2331)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2315..2331)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2762..3553
                     /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      3646..4234
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4135..4154
                     /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 atggatacca acgggtctac ggagggatcg cggtcgcggt cgcgatcttt
      721 agacatacag cccagctccg aaggactggg gcccacttcg gaaccgtttc cttcttcaga
      781 tgacagtccc aggtcggccc tggcagctgc aaccgcagca gctgcagcgg ctgcatcagc
      841 tgctgcagct actgcagcct tcaccactgc caaagcagct gcattatcta caaagacccc
      901 agcgccctgt tctgagttca tggagccgtc ctctgacccc agccttcttg gggagccctg
      961 tgcgggaccc ggctttaccc acaatatagc ccatgggagt cttggctttg agcccgtcta
     1021 tgtttcctgt attgctcagg acacttgcac tacaactgac catagttcta atcctggccc
     1081 tgttccaggc tctagctctg ggcctgttct tggttccagc tcaggtgctg gccatggctc
     1141 tggctctggc tctggtcctg gctgtggctc tgtccctggc tctggctctg gtcctggtcc
     1201 tggctctggt cctggctctg gtcctggtca tggctctggc tctcatcctg gtcctgcctc
     1261 tgggcctggt ccagacactg gccctgactc tgagctcagc ccctgtattc ctccagggtt
     1321 cagaaacctg gtggcagatc gggtccttaa ctatacctcc tggagtcagc actgcccctg
     1381 ggagccccag aaacaaccac cttgggaatt tttgcaagtc ttagaaccgg gtgcccgagg
     1441 attatggaaa cccccagaca ttaaagggaa gcttatggtt tgctatgaaa ctttgccgcg
     1501 gggccagtgc ctcctctaca actgggagga agagagagcc accaaccacc tggatcaagt
     1561 cccaagcatg caggatggct ctgagagttt tttcttccga cacggacacc ggggactgct
     1621 gactatgcaa ctaaagtcac ccatgccctc cagcaccacc cagaaagact cgtaccagcc
     1681 accaggaaac gtctattggc tacttcgagg gaagcgtgaa gccatgctgg agatgctcct
     1741 gcagcatcag atctgtaaag aggtgcaggc agaacaggaa cccacaagga agctcttcga
     1801 ggttgagtct gtgacacacc atgactaccg aatggagctg gcacaagcag ggactcctgc
     1861 cccaacaaag cctcacgact accgccagga gcaacctgag accttctgga tacagagggc
     1921 accacagctg ccgacatggt ggccattgcc cacccaggta ccagcagcag aagactatct
     1981 gacttggaaa gaatgggggt ttacaggagt ccaggaggtc ctttccgctc tcctaagagc
     2041 cacacctggt gaatactcag taaacatttg cggaatgaat gaacaccctg tctgcagcag
     2101 gacatggaca aataggctct gccatcagga aatgggaagc aagaaaacgg taactcaaga
     2161 ggacagaggc tggtgcccaa ctttcttgta caaagttggc attataagaa agcattgctt
     2221 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     2281 agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct ggcagctctg
     2341 gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgcct
     2401 cctctagacc agccaggaca gaaatgcctc gacttcgctg ctacccaagg ttgccgggtg
     2461 acgcacaccg tggaaacgga tgaaggcacg aacccagtgg acataagcct gttcggttcg
     2521 taagctgtaa tgcaagtagc gtatgcgctc acgcaactgg tccagaacct tgaccgaacg
     2581 cagcggtggt aacggcgcag tggcggtttt catggcttgt tatgactgtt tttttggggt
     2641 acagtctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat
     2701 gttatggagc agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaaacat
     2761 tatgagggaa gcggtgatcg ccgaagtatc gactcaacta tcagaggtag ttggcgtcat
     2821 cgagcgccat ctcgaaccga cgttgctggc cgtacatttg tacggctccg cagtggatgg
     2881 cggcctgaag ccacacagtg atattgattt gctggttacg gtgaccgtaa ggcttgatga
     2941 aacaacgcgg cgagctttga tcaacgacct tttggaaact tcggcttccc ctggagagag
     3001 cgagattctc cgcgctgtag aagtcaccat tgttgtgcac gacgacatca ttccgtggcg
     3061 ttatccagct aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg
     3121 tatcttcgag ccagccacga tcgacattga tctggctatc ttgctgacaa aagcaagaga
     3181 acatagcgtt gccttggtag gtccagcggc ggaggaactc tttgatccgg ttcctgaaca
     3241 ggatctattt gaggcgctaa atgaaacctt aacgctatgg aactcgccgc ccgactgggc
     3301 tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg
     3361 caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc cggcccagta
     3421 tcagcccgtc atacttgaag ctagacaggc ttatcttgga caagaagaag atcgcttggc
     3481 ctcgcgcgca gatcagttgg aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt
     3541 agtcggcaaa taaccctcga gccacccatg accaaaatcc cttaacgtga gttacgcgtc
     3601 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3661 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3721 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3781 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3841 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3901 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3961 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4021 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4081 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     4141 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     4201 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     4261 gttcctggcc ttttgctggc cttttgctca catgtt
//
