LOCUS       sequence-164911-        4428 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing PAK1 , part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4428)
  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 4428)
  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_PAK1_WT
FEATURES             Location/Qualifiers
     source          1..4428
                     /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"
     CDS             complement(724..741)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(1828..1927)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1944..1963)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1945..1963)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1968..1984)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1968..1984)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2415..3206
                     /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      3299..3887
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3788..3807
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4041..4058
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4217..4244)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4336..4422)
                     /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 catgtcaaat aacggcctag acattcaaga caaaccccca gcccctccga
      241 tgagaaatac cagcactatg attggagccg gcagcaaaga tgctggaacc ctaaaccatg
      301 gttctaaacc tctgcctcca aacccagagg agaagaaaaa gaaggaccga ttttaccgat
      361 ccattttacc tggagataaa acaaataaaa agaaagagaa agagcggcca gagatttctc
      421 tcccttcaga ttttgaacac acaattcatg tcggttttga tgctgtcaca ggggagttta
      481 cgggaatgcc agagcagtgg gcccgcttgc ttcagacatc aaatatcact aagtcggagc
      541 agaagaaaaa cccgcaggct gttctggatg tgttggagtt ttacaactcg aagaagacat
      601 ccaacagcca gaaatacatg agctttacag ataagtcagc tgaggattac aattcttcta
      661 atgccttgaa tgtgaaggct gtgtctgaga ctcctgcagt gccaccagtt tcagaagatg
      721 aggatgatga tgatgatgat gctaccccac caccagtgat tgctccacgc ccagagcaca
      781 caaaatctgt atacacacgg tctgtgattg aaccacttcc tgtcactcca actcgggacg
      841 tggctacatc tcccatttca cctactgaaa ataacaccac tccaccagat gctttgaccc
      901 ggaatactga gaagcagaag aagaagccta aaatgtctga tgaggagatc ttggagaaat
      961 tacgaagcat agtgagtgtg ggcgatccta agaagaaata tacacggttt gagaagattg
     1021 gacaaggtgc ttcaggcacc gtgtacacag caatggatgt ggccacagga caggaggtgg
     1081 ccattaagca gatgaatctt cagcagcagc ccaagaaaga gctgattatt aatgagatcc
     1141 tggtcatgag ggaaaacaag aacccaaaca ttgtgaatta cttggacagt tacctcgtgg
     1201 gagatgagct gtgggttgtt atggaatact tggctggagg ctccttgaca gatgtggtga
     1261 cagaaacttg catggatgaa ggccaaattg cagctgtgtg ccgtgagtgt ctgcaggctc
     1321 tggagttctt gcattcgaac caggtcattc acagagacat caagagtgac aatattctgt
     1381 tgggaatgga tggctctgtc aagctaactg actttggatt ctgtgcacag ataaccccag
     1441 agcagagcaa acggagcacc atggtaggaa ccccatactg gatggcacca gaggttgtga
     1501 cacgaaaggc ctatgggccc aaggttgaca tctggtccct gggcatcatg gccatcgaaa
     1561 tgattgaagg ggagcctcca tacctcaatg aaaaccctct gagagccttg tacctcattg
     1621 ccaccaatgg gaccccagaa cttcagaacc cagagaagct gtcagctatc ttccgggact
     1681 ttctgaaccg ctgtctcgag atggatgtgg agaagagagg ttcagctaaa gagctgctac
     1741 agcatcaatt cctgaagatt gccaagcccc tctccagcct cactccactg attgctgcag
     1801 ctaaggaggc aacaaagaac aatcacttgc caactttctt gtacaaagtt ggcattataa
     1861 gaaagcattg cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca
     1921 ttatttgcca tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt
     1981 cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata
     2041 tatcatcatg cctcctctag accagccagg acagaaatgc ctcgacttcg ctgctaccca
     2101 aggttgccgg gtgacgcaca ccgtggaaac ggatgaaggc acgaacccag tggacataag
     2161 cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa
     2221 ccttgaccga acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact
     2281 gtttttttgg ggtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg
     2341 gtcgatgttt gatgttatgg agcagcaacg atgttacgca gcagggcagt cgccctaaaa
     2401 caaagttaaa cattatgagg gaagcggtga tcgccgaagt atcgactcaa ctatcagagg
     2461 tagttggcgt catcgagcgc catctcgaac cgacgttgct ggccgtacat ttgtacggct
     2521 ccgcagtgga tggcggcctg aagccacaca gtgatattga tttgctggtt acggtgaccg
     2581 taaggcttga tgaaacaacg cggcgagctt tgatcaacga ccttttggaa acttcggctt
     2641 cccctggaga gagcgagatt ctccgcgctg tagaagtcac cattgttgtg cacgacgaca
     2701 tcattccgtg gcgttatcca gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg
     2761 acattcttgc aggtatcttc gagccagcca cgatcgacat tgatctggct atcttgctga
     2821 caaaagcaag agaacatagc gttgccttgg taggtccagc ggcggaggaa ctctttgatc
     2881 cggttcctga acaggatcta tttgaggcgc taaatgaaac cttaacgcta tggaactcgc
     2941 cgcccgactg ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc atttggtaca
     3001 gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca atggagcgcc
     3061 tgccggccca gtatcagccc gtcatacttg aagctagaca ggcttatctt ggacaagaag
     3121 aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt tgtccactac gtgaaaggcg
     3181 agatcaccaa ggtagtcggc aaataaccct cgagccaccc atgaccaaaa tcccttaacg
     3241 tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3301 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3361 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3421 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     3481 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3541 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3601 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3661 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3721 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3781 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3841 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3901 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     3961 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     4021 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     4081 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     4141 ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag
     4201 agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct
     4261 ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa
     4321 atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa
     4381 acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcct
//
