LOCUS       sequence-165724-        6870 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing PIK3R4 ,part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6870)
  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 6870)
  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_PIK3R4_p.E1209K
FEATURES             Location/Qualifiers
     source          1..6870
                     /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(4270..4369)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4386..4405)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4387..4405)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4410..4426)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4410..4426)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4857..5648
                     /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      5741..6329
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6230..6249
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6483..6500
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(6659..6686)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(6778..6864)
                     /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 catgggaaat cagcttgctg gcattgctcc ctcccagatc ctttctgtag
      241 agagttattt ttcagatatt catgactttg aatatgataa aagcctgggg agtactcggt
      301 tttttaaagt tgctcgagcc aagcaccgag aaggcctggt cgttgtgaag gtttttgcaa
      361 ttcaggatcc cacattgcct ttaaccagct ataaacaaga gctggaggaa ctgaaaatca
      421 ggcttaattc tgcacagaat tgtctacctt tccagaaagc atcagaaaaa gcatctgaga
      481 aagcagctat gctctttagg cagtatgtgc gagacaatct ctatgatcgc atcagtaccc
      541 gtccattctt gaataacatt gagaagcgct ggattgcttt ccagatcctg acagctgtgg
      601 accaagcaca caaatctgga gttcgtcatg gggacatcaa gactgagaat gtgatggtca
      661 ccagttggaa ttgggttctt ctaactgatt ttgccagttt taagcccact tatcttccag
      721 aagacaaccc ggcagatttc aattatttct ttgacacatc acggaggaga acttgctata
      781 ttgctcctga acgttttgtt gatggtggga tgtttgccac tgagttagaa tatatgagag
      841 atccttcaac tccgcttgta gacttaaata gcaatcagag aagaagagga gagttgaaga
      901 gagcaatgga catcttttca gcaggttgtg tgatagctga gctttttaca gaaggtgtac
      961 cattatttga tctctctcaa cttttggctt atagaaatgg acattttttc cctgaacaag
     1021 tgctaaataa aattgaagat cacagtatca gagaattggt aactcagatg attcaccgtg
     1081 agccagataa acgtttagag gcagaagatt acttaaaaca gcagcgtggc aatgcctttc
     1141 ctgaaatatt ttacactttt cttcagccct acatggccca gtttgccaag gaaacgtttc
     1201 tttctgcaga tgagcgtatt ctggttatac ggaaggattt gggcaacatt attcacaatc
     1261 tctgtggaca tgatctgcca gaaaaagccg aaggagagcc taaggaaaat gggctggtta
     1321 tcttggtatc tgttataaca tcctgcctac agacccttaa atactgtgat tccaaactag
     1381 ctgctttgga actgattctt catttggctc caagattaag tgttgaaatc cttttggatc
     1441 gtattactcc atatcttttg catttcagca atgactctgt tcctagggtg agggctgaag
     1501 ccttgaggac gttgaccaaa gttcttgctc tcgtcaaaga ggttcctcgt aatgatatca
     1561 atatttatcc ggaatacatt ctgccaggca tagcccactt agcccaagat gatgctacta
     1621 tcgttagact agcctatgct gaaaacatag ctctgctggc agaaacagct ctgagattcc
     1681 tggaattagt acagttaaaa aatcttaata tggaaaatga ccccaataat gaagaaatag
     1741 atgaggttac acatccaaat ggaaattatg acacagagct ccaagcctta catgaaatgg
     1801 tccagcagaa agttgttact ttgctaagtg accctgaaaa tattgtaaaa caaaccttga
     1861 tggaaaatgg aataacacgg ctgtgtgtat tctttggacg tcagaaagcc aacgatgttt
     1921 tgttgtccca catgattact ttcctaaatg ataagaatga ttggcatcta cgtggagcat
     1981 tttttgatag tatagttggt gttgctgcct atgttggctg gcaaagctcc tcaattctca
     2041 agcctctgct gcaacaaggt cttagtgatg ctgaggaatt tgtcattgtg aaagctcttt
     2101 atgcccttac ttgtatgtgc cagttaggac tgctacaaaa accccatgtt tacgaatttg
     2161 ccagtgacat tgcccccttc ctgtgtcatc ccaatttatg gatacgttat ggtgccgtgg
     2221 gatttatcac agtggtagct cgtcaaataa gtacagctga tgtctactgt aaactgatgc
     2281 cttatcttga cccatatatt acccaaccaa taatacagat tgaaagaaaa cttgttctgc
     2341 tcagtgtttt aaaggaacca gtaagtcgtt ctatatttga ttatgctttg aggtctaaag
     2401 atattactag cttgttcaga catcttcaca tgcgtcagaa gaaacgaaat ggttctcttc
     2461 ccgactgccc tccgccagag gatcctgcca tagcacagct tctgaagaag ttgctctcac
     2521 agggaatgac agaggaagag gaagacaaac ttctggcact gaaagacttc atgatgaaat
     2581 ctaataaagc aaaggccaat atagtggacc agagccatct tcatgatagt agtcagaaag
     2641 gtgtaattga cttggcagct ttaggcataa ctgggagaca agttgatctt gttaaaacca
     2701 aacaagaacc agatgacaaa cgggccagaa aacatgtaaa acaagactca aatgtaaatg
     2761 aagaatggaa aagcatgttt gggtcactgg acccaccaaa catgccacag gccctaccta
     2821 aagggagtga tcaggaggtg attcagactg ggaaacctcc tcgttccgag tcctctgctg
     2881 gcatttgtgt ccctttgtca acttcttcac aggttccaga agtgacaact gtccaaaata
     2941 aaaaaccagt aataccggtt ttaagtagta caatcttacc atccacctat cagattcgaa
     3001 ttacaacttg taaaactgaa cttcagcaac tcatccagca aaagcgggag cagtgcaatg
     3061 ctgagagaat agctaagcag atgatggaaa atgctgaatg ggagagtaaa ccaccaccac
     3121 ctggatggcg tcctaaaggg ctgttagttg cccatcttca tgagcataaa tctgctgtga
     3181 atcgaattag agtctctgat gaacactcac tttttgcaac atgttcaaat gatggcacag
     3241 tgaaaatctg gaacagtcaa aagatggagg ggaagaccac cactaccaga tctattctta
     3301 catacagccg aattggagga cgagtcaaga cgctcacatt ctgccaaggc tcccactatt
     3361 tagccatagc atctgataat ggtgctgtcc agcttcttgg aattgaggct tctaagctgc
     3421 ccaagtctcc taaaatccat cctctacaaa gcagaattct agatcagaag gaggacggtt
     3481 gtgttgtgga tatgcatcac ttcaactctg gagcacagtc tgttcttgcc tatgccactg
     3541 tgaatggctc tctggttggc tgggacctta ggtcttcaag caatgcgtgg actttaaagc
     3601 atgatttaaa gtcgggcctc atcacttcct ttgctgtgga catccaccaa tgctggctct
     3661 gcattggtac aagcagtggt accatggctt gttgggacat gaggttccag ttgccaattt
     3721 caagtcactg tcatccttcc agggctcgaa tcagacgcct ctcaatgcac cctctgtatc
     3781 agtcctgggt gattgcagct gttcagggca acaacaaagt gtccatgtgg gacatggaga
     3841 ctggtgacag aagatttact ctctgggcca gcagtgcacc accactttct gaattacagc
     3901 cttctcctca tagcgtccat ggtatctact gtagtcctgc agatggaaat cctatcctac
     3961 taacagctgg ctcagatatg aaaataaggt tttgggactt ggcttaccca gaaaggtcct
     4021 atgttgttgc aggaagtact agttccccat ctgtgtccta ctacaggaaa ataattgaag
     4081 gcactgaagt tgtccaggaa attcagaata agcagaaagt aggaccaagt gatgacaccc
     4141 ctcgaagggg cccagagtcc ctgcccgtgg gacatcatga catcatcact gatgtcgcca
     4201 cattccagac cacacagggc ttcatcgtaa ctgcttctag agatgggatt gtgaaggtgt
     4261 ggaaatgatt gccaactttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa
     4321 tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg
     4381 atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg
     4441 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgcctcctct
     4501 agaccagcca ggacagaaat gcctcgactt cgctgctacc caaggttgcc gggtgacgca
     4561 caccgtggaa acggatgaag gcacgaaccc agtggacata agcctgttcg gttcgtaagc
     4621 tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc gaacgcagcg
     4681 gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt ggggtacagt
     4741 ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat
     4801 ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta aacattatga
     4861 gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc gtcatcgagc
     4921 gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg gatggcggcc
     4981 tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa
     5041 cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga gagagcgaga
     5101 ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg tggcgttatc
     5161 cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt gcaggtatct
     5221 tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca agagaacata
     5281 gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct gaacaggatc
     5341 tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac tgggctggcg
     5401 atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta accggcaaaa
     5461 tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc cagtatcagc
     5521 ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc ttggcctcgc
     5581 gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc aaggtagtcg
     5641 gcaaataacc ctcgagccac ccatgaccaa aatcccttaa cgtgagttac gcgtcgttcc
     5701 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5761 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5821 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5881 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5941 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6001 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6061 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6121 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6181 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6241 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6301 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6361 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     6421 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     6481 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     6541 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     6601 gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga agagtttgta gaaacgcaaa
     6661 aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt atggcgggcg
     6721 tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc ccggcggatt
     6781 tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttc
     6841 cgactgagcc tttcgtttta tttgatgcct
//
