LOCUS       sequence-165298-        4512 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing PRKAA1, part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4512)
  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 4512)
  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_PRKAA1_WT
FEATURES             Location/Qualifiers
     source          1..4512
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     469..488
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     722..739
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(898..925)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1017..1103)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1152..1174
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1166..1183
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1167..1183
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1199..1298
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3021..3120)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3137..3156)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3138..3156)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3161..3177)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3161..3177)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3608..4399
                     /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      join(4492..4512,1..568)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
       61 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
      121 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
      181 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
      241 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
      301 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
      361 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
      421 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
      481 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
      541 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
      601 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
      661 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
      721 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
      781 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
      841 tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa
      901 aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt
      961 cctgcccgcc accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt
     1021 gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc
     1081 gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc
     1141 atggatgttt tcccagtcac gacgttgtaa aacgacggcc agtcttaagc tcgggcccca
     1201 aataatgatt ttattttgac tgatagtgac ctgttcgttg caacaaattg atgagcaatg
     1261 cttttttata atgccaactt tgtacaaaaa agttggcatg cgcagactca gttcctggag
     1321 aaagttggcg acagccgaga agcagaaaca cgacgggcgg gtgaagatcg gccactacat
     1381 tctgggtgac acgctggggg tcggcacctt cggcaaagtg aaggttggca aacatgaatt
     1441 gactgggcat aaagtagctg tgaagatact caatcgacag aagattcgga gccttgatgt
     1501 ggtaggaaaa atccgcagag aaattcagaa cctcaagctt ttcaggcatc ctcatataat
     1561 taaactgtac caggtcatca gtacaccatc tgatattttc atggtgatgg aatatgtctc
     1621 aggaggagag ctatttgatt atatctgtaa gaatggaagg aaatctgatg tacctggagt
     1681 agtaaaaaca ggctccacga aggagctgga tgaaaaagaa agtcggcgtc tgttccaaca
     1741 gatcctttct ggtgtggatt attgtcacag gcatatggtg gtccatagag atttgaaacc
     1801 tgaaaatgtc ctgcttgatg cacacatgaa tgcaaagata gctgattttg gtctttcaaa
     1861 catgatgtca gatggtgaat ttttaagaac aagttgtggc tcacccaact atgctgcacc
     1921 agaagtaatt tcaggaagat tgtatgcagg cccagaggta gatatatgga gcagtggggt
     1981 tattctctat gctttattat gtggaaccct tccatttgat gatgaccatg tgccaactct
     2041 ttttaagaag atatgtgatg ggatcttcta tacccctcaa tatttaaatc cttctgtgat
     2101 tagccttttg aaacatatgc tgcaggtgga tcccatgaag agggccacaa tcaaagatat
     2161 cagggaacat gaatggttta aacaggacct tccaaaatat ctctttcctg aggatccatc
     2221 atatagttca accatgattg atgatgaagc cttaaaagaa gtatgtgaaa agtttgagtg
     2281 ctcagaagag gaagttctca gctgtcttta caacagaaat caccaggatc ctttggcagt
     2341 tgcctaccat ctcataatag ataacaggag aataatgaat gaagccaaag atttctattt
     2401 ggcgacaagc ccacctgatt cttttcttga tgatcatcac ctgactcggc cccatcctga
     2461 aagagtacca ttcttggttg ctgaaacacc aagggcacgc catacccttg atgaattaaa
     2521 tccacagaaa tccaaacacc aaggtgtaag gaaagcaaaa tggcatttag gaattagaag
     2581 tcaaagtcga ccaaatgata ttatggcaga agtatgtaga gcaatcaaac aattggatta
     2641 tgaatggaag gttgtaaacc catattattt gcgtgtacga aggaagaatc ctgtgacaag
     2701 cacttactcc aaaatgagtc tacagttata ccaagtggat agtagaactt atctactgga
     2761 tttccgtagt attgatgatg aaattacaga agccaaatca gggactgcta ctccacagag
     2821 atcgggatca gttagcaact atcgatcttg ccaaaggagt gattcagatg ctgaggctca
     2881 aggaaaatcc tcagaagttt ctcttacctc atctgtgacc tcacttgact cttctcctgt
     2941 tgacctaact ccaagacctg gaagtcacac aatagaattt tttgagatgt gtgcaaatct
     3001 aattaaaatt cttgcacaat acccaacttt cttgtacaaa gttggcatta taagaaagca
     3061 ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg
     3121 ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca
     3181 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     3241 atgcctcctc tagaccagcc aggacagaaa tgcctcgact tcgctgctac ccaaggttgc
     3301 cgggtgacgc acaccgtgga aacggatgaa ggcacgaacc cagtggacat aagcctgttc
     3361 ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac
     3421 cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt
     3481 tggggtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg
     3541 tttgatgtta tggagcagca acgatgttac gcagcagggc agtcgcccta aaacaaagtt
     3601 aaacattatg agggaagcgg tgatcgccga agtatcgact caactatcag aggtagttgg
     3661 cgtcatcgag cgccatctcg aaccgacgtt gctggccgta catttgtacg gctccgcagt
     3721 ggatggcggc ctgaagccac acagtgatat tgatttgctg gttacggtga ccgtaaggct
     3781 tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg cttcccctgg
     3841 agagagcgag attctccgcg ctgtagaagt caccattgtt gtgcacgacg acatcattcc
     3901 gtggcgttat ccagctaagc gcgaactgca atttggagaa tggcagcgca atgacattct
     3961 tgcaggtatc ttcgagccag ccacgatcga cattgatctg gctatcttgc tgacaaaagc
     4021 aagagaacat agcgttgcct tggtaggtcc agcggcggag gaactctttg atccggttcc
     4081 tgaacaggat ctatttgagg cgctaaatga aaccttaacg ctatggaact cgccgcccga
     4141 ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt acagcgcagt
     4201 aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg gcaatggagc gcctgccggc
     4261 ccagtatcag cccgtcatac ttgaagctag acaggcttat cttggacaag aagaagatcg
     4321 cttggcctcg cgcgcagatc agttggaaga atttgtccac tacgtgaaag gcgagatcac
     4381 caaggtagtc ggcaaataac cctcgagcca cccatgacca aaatccctta acgtgagtta
     4441 cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4501 tttttttctg cg
//
