LOCUS       sequence-169159-        4359 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing PNKP , part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4359)
  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 4359)
  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_PNKP_WT
FEATURES             Location/Qualifiers
     source          1..4359
                     /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(2238..2337)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2354..2373)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2355..2373)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2378..2394)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2378..2394)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2825..3616
                     /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      3709..4297
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4198..4217
                     /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 atgggcgagg tggaggcccc gggccgcttg tggctcgaga gcccccctgg
      721 gggagcgccc cccatcttcc tgccctcgga cgggcaagcc ctggtcctgg gcaggggacc
      781 cctgacccag gttacggacc ggaagtgctc cagaactcaa gtggagctgg tcgcagatcc
      841 tgagacccgg acagtggcag tgaaacagct gggagttaac ccctcaacta ccgggaccca
      901 ggagttgaag ccggggttgg agggctctct gggggtgggg gacacactgt atttggtcaa
      961 tggcctccac ccactgaccc tgcgctggga agagacccgc acaccagaat cccagccaga
     1021 tactccgcct ggcacccctc tggtgtccca agatgagaag agagatgctg agctgccgaa
     1081 gaagcgtatg cggaagtcaa accccggctg ggagaacttg gagaagttgc tagtgttcac
     1141 cgcagctggg gtgaaacccc agggcaaggt ggctggcttt gatctggacg ggacgctcat
     1201 caccacacgc tctgggaagg tctttcccac tggccccagt gactggagga tcttgtaccc
     1261 agagattccc cgtaagctcc gagagctgga agccgagggc tacaagctgg tgatcttcac
     1321 caaccagatg agcatcgggc gcgggaagct gccagccgag gagttcaagg ccaaggtgga
     1381 ggctgtggtg gagaagctgg gggtcccctt ccaggtgctg gtggccacgc acgcaggctt
     1441 gtaccggaag ccggtgacgg gcatgtggga ccatctgcag gagcaggcca acgacggcac
     1501 gcccatatcc atcggggaca gcatctttgt gggagacgca gccggacgcc cggccaactg
     1561 ggccccgggg cggaagaaga aagacttctc ctgcgccgat cgcctgtttg ccctcaacct
     1621 tggcctgccc ttcgccacgc ctgaggagtt ctttctcaag tggccagcag ccggcttcga
     1681 gctcccagcc tttgatccga ggactgtctc ccgctcaggg cctctctgcc tccccgagtc
     1741 cagggccctc ctgagcgcca gcccggaggt ggttgtcgca gtgggattcc ctggggccgg
     1801 gaagtccacc tttctcaaga agcacctcgt gtcggccgga tatgtccacg tgaacaggga
     1861 cacgctaggc tcctggcagc gctgtgtgac cacgtgtgag acagccctga agcaagggaa
     1921 acgggtcgcc atcgacaaca caaacccaga cgccgcgagc cgcgccaggt acgtccagtg
     1981 tgcccgagcc gcgggcgtcc cctgccgctg cttcctcttc accgccactc tggagcaggc
     2041 gcgccacaac aaccggtttc gagagatgac ggactcctct catatccccg tgtcagacat
     2101 ggtcatgtat ggctacagga agcagttcga ggccccaacg ctggctgaag gcttctctgc
     2161 catcctggag atcccgttcc ggctatgggt ggagccgagg ctggggcggc tgtactgcca
     2221 gttctccgag ggctgattgc caactttctt gtacaaagtt ggcattataa gaaagcattg
     2281 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
     2341 tccagctgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
     2401 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
     2461 cctcctctag accagccagg acagaaatgc ctcgacttcg ctgctaccca aggttgccgg
     2521 gtgacgcaca ccgtggaaac ggatgaaggc acgaacccag tggacataag cctgttcggt
     2581 tcgtaagctg taatgcaagt agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga
     2641 acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact gtttttttgg
     2701 ggtacagtct atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt
     2761 gatgttatgg agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttaaa
     2821 cattatgagg gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt
     2881 catcgagcgc catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga
     2941 tggcggcctg aagccacaca gtgatattga tttgctggtt acggtgaccg taaggcttga
     3001 tgaaacaacg cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga
     3061 gagcgagatt ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg
     3121 gcgttatcca gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc
     3181 aggtatcttc gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag
     3241 agaacatagc gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga
     3301 acaggatcta tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg
     3361 ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac
     3421 cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca
     3481 gtatcagccc gtcatacttg aagctagaca ggcttatctt ggacaagaag aagatcgctt
     3541 ggcctcgcgc gcagatcagt tggaagaatt tgtccactac gtgaaaggcg agatcaccaa
     3601 ggtagtcggc aaataaccct cgagccaccc atgaccaaaa tcccttaacg tgagttacgc
     3661 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3721 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3781 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3841 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3901 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3961 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4021 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4081 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4141 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4201 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4261 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4321 acggttcctg gccttttgct ggccttttgc tcacatgtt
//
