LOCUS       sequence-169269-        3288 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing PPIAL4G , part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3288)
  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 3288)
  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_PPIAL4G_WT
FEATURES             Location/Qualifiers
     source          1..3288
                     /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(1167..1266)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1283..1302)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1284..1302)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1307..1323)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1307..1323)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1754..2545
                     /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      2638..3226
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3127..3146
                     /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 atgacaactt tgtacaaaaa
      661 agttggcacc atggtcaact ccgtcgtctt ttttgacatc accgtcgacg gcaagccctt
      721 gggccgcatc tccatcaaac tgtttgcaga caagattcca aagacagcag aaaactttcg
      781 tgctctgagc actggagaga aaggatttcg ttataagggt tcctgctttc acagaattat
      841 tccagggttt atgtgtcagg gtggtgactt cacacgccct aatggcaccg gtgacaagtc
      901 catctatggg gagaaatttg atgatgagaa cctcatccga aagcatacag gttctggcat
      961 cttgtccatg gcaaatgctg gacccaacac aaatggttcc cagtttttca tctgcactgc
     1021 caagactgag tggttggatg gcaagcatgt ggcctttggc aaggtgaaag aacgtgtgaa
     1081 tattgtggaa gccatggagc actttgggta caggaatagc aagaccagca agaagatcac
     1141 cattgctgac tgtggacaat tctgattgcc aactttcttg tacaaagttg tcattataag
     1201 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
     1261 tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc
     1321 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     1381 atcatcatgc ctcctctaga ccagccagga cagaaatgcc tcgacttcgc tgctacccaa
     1441 ggttgccggg tgacgcacac cgtggaaacg gatgaaggca cgaacccagt ggacataagc
     1501 ctgttcggtt cgtaagctgt aatgcaagta gcgtatgcgc tcacgcaact ggtccagaac
     1561 cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt gttatgactg
     1621 tttttttggg gtacagtcta tgcctcgggc atccaagcag caagcgcgtt acgccgtggg
     1681 tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagggcagtc gccctaaaac
     1741 aaagttaaac attatgaggg aagcggtgat cgccgaagta tcgactcaac tatcagaggt
     1801 agttggcgtc atcgagcgcc atctcgaacc gacgttgctg gccgtacatt tgtacggctc
     1861 cgcagtggat ggcggcctga agccacacag tgatattgat ttgctggtta cggtgaccgt
     1921 aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa cttcggcttc
     1981 ccctggagag agcgagattc tccgcgctgt agaagtcacc attgttgtgc acgacgacat
     2041 cattccgtgg cgttatccag ctaagcgcga actgcaattt ggagaatggc agcgcaatga
     2101 cattcttgca ggtatcttcg agccagccac gatcgacatt gatctggcta tcttgctgac
     2161 aaaagcaaga gaacatagcg ttgccttggt aggtccagcg gcggaggaac tctttgatcc
     2221 ggttcctgaa caggatctat ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc
     2281 gcccgactgg gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca tttggtacag
     2341 cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa tggagcgcct
     2401 gccggcccag tatcagcccg tcatacttga agctagacag gcttatcttg gacaagaaga
     2461 agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gtccactacg tgaaaggcga
     2521 gatcaccaag gtagtcggca aataaccctc gagccaccca tgaccaaaat accttaacgt
     2581 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2641 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2701 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2761 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     2821 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2881 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2941 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3001 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3061 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3121 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3181 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3241 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtt
//
