LOCUS       sequence-162921-        5334 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing PYGL, used as an over expression 
            control for gene expression analysis. Part of the Target Accelerator
            Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5334)
  AUTHORS   Berger AH, Brooks AN, Wu X, Shrestha Y, Chouinard C, Piccioni F, 
            Bagul M, Kamburov A, Imielinski M, Hogstrom L, Zhu C, Yang X, Pantel
            S, Sakai R, Watson J, Kaplan N, Campbell JD, Singh S, Root DE, 
            Narayan R, Natoli T, Lahr DL, Tirosh I, Tamayo P, Getz G, Wong B, 
            Doench J, Subramanian A, Golub TR, Meyerson M, Boehm JS
  TITLE     High-throughput Phenotyping of Lung Cancer Somatic Mutations.
  JOURNAL   Cancer Cell. 2016 Aug 8;30(2):214-28. doi: 
            10.1016/j.ccell.2016.06.022. Epub 2016 Jul 28.
  PUBMED    27478040
REFERENCE   2  (bases 1 to 5334)
  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: 
            "10.1016/j.ccell.2016.06.022"; journalName: "Cancer Cell"; date: 
            "2016-08-8- 8"; volume: "30"; issue: "2"; pages: "214-28"
COMMENT     Sequence Label: pDONR223_PYGL_WT_V5
FEATURES             Location/Qualifiers
     source          1..5334
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(100..117)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(271..859)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(351..370)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(952..1743)
                     /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"
     primer_bind     2174..2190
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2174..2190
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2195..2214
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2195..2213
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2231..2330
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(4875..4974)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4990..5007)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4990..5006)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4999..5021)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      5070..5156
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5248..5275
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
       61 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
      121 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
      181 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
      241 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
      301 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
      361 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
      421 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
      481 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
      541 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
      601 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
      661 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
      721 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
      781 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
      841 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
      901 gaacgacgcg taactcacgt taagggattt tggtcatggg tggctcgagg gttatttgcc
      961 gactaccttg gtgatctcgc ctttcacgta gtggacaaat tcttccaact gatctgcgcg
     1021 cgaggccaag cgatcttctt cttgtccaag ataagcctgt ctagcttcaa gtatgacggg
     1081 ctgatactgg gccggcaggc gctccattgc ccagtcggca gcgacatcct tcggcgcgat
     1141 tttgccggtt actgcgctgt accaaatgcg ggacaacgta agcactacat ttcgctcatc
     1201 gccagcccag tcgggcggcg agttccatag cgttaaggtt tcatttagcg cctcaaatag
     1261 atcctgttca ggaaccggat caaagagttc ctccgccgct ggacctacca aggcaacgct
     1321 atgttctctt gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa
     1381 gatacctgca agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg
     1441 ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg acttctacag cgcggagaat
     1501 ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt
     1561 tgtttcatca agccttacgg tcaccgtaac cagcaaatca atatcactgt gtggcttcag
     1621 gccgccatcc actgcggagc cgtacaaatg tacggccagc aacgtcggtt cgagatggcg
     1681 ctcgatgacg ccaactacct ctgatagttg agtcgatact tcggcgatca ccgcttccct
     1741 cataatgttt aactttgttt tagggcgact gccctgctgc gtaacatcgt tgctgctcca
     1801 taacatcaaa catcgaccca cggcgtaacg cgcttgctgc ttggatgccc gaggcataga
     1861 ctgtacccca aaaaaacagt cataacaagc catgaaaacc gccactgcgc cgttaccacc
     1921 gctgcgttcg gtcaaggttc tggaccagtt gcgtgagcgc atacgctact tgcattacag
     1981 cttacgaacc gaacaggctt atgtccactg ggttcgtgcc ttcatccgtt tccacggtgt
     2041 gcgtcacccg gcaaccttgg gtagcagcga agtcgaggca tttctgtcct ggctggtcta
     2101 gaggaggcat gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac
     2161 gggccagagc tgccaggaaa cagctatgac catgtaatac gactcactat aggggatatc
     2221 agctggatgg caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat
     2281 tgataagcaa tgctttctta taatgccaac tttgtacaag aaagttggca aatttccatt
     2341 gactttgtta gattcattgg atagagaaat ctttagatct gaaggttcca cgttccagat
     2401 gttttgggca tattctttaa ttgttcggtc actggagaat ttccccgagg cagctatgtt
     2461 tttgagtacc attgtgttcc aggcctttgg attcatgtac agctgactca ctttatcttg
     2521 acacttgaca taggcttcgt agtctgcaaa gactttaaac ctgtcatgat aaaatagcat
     2581 gttgatgata tctttgaaga ggtcaggctg cttgggagaa aaaaagccat tgtcaatttg
     2641 atcaatgacc agcttcagct ctggaagtgc ctcatagtat tcttttgcct cgtacccttt
     2701 cttgtccaaa gcagccacat catctatcct catgccaaag atgaacaggt tctcttcccc
     2761 agcttcttct gccatttcca cattggcccc atccatggtc ccgatagtta gggccccatt
     2821 tagcatgaac ttcatattgc ctgtccccga ggcttcggtg cctgcagtgg aaatctgctc
     2881 tgacagatct gtggctggaa tgactttttc agcaagagat actctgtagt tctccaagaa
     2941 gatgactttc aacttgcttc caaccatagg gtcattgttc accacatctg ccactgaagt
     3001 gatcagcttt atgatcattt tggccatgtg atatcctggg gcagctttac caccaatgat
     3061 aactgtcctt ggcacgaata acttcttagg gtctttctta atgcggttgt acatcgtgat
     3121 cacatgcaga cagttcaaga gctgtcgctt gtactcatgt atcctcttca cctggacatc
     3181 aaacatggag gatgggttga tcttcacttt gtactccgtc tccaggaact gagaaaactt
     3241 cagcttattc tcctgcttca ccttggcgag ttcccggagg aagacatcat cacccaggaa
     3301 gctgtggagc ttcgtcagct ggctcaggtc tttcacatag tcttctccaa ttttctctgc
     3361 tatgagctct gcaagtcctg ggttgcagag taggagccag cgccttggag tgatcccatt
     3421 ggttttattc tgaaacttgt caggttctag ctcactgaag tccttgaata ctttagtctt
     3481 cacgatgtct gagtggattt tagccacgcc attcacagca tgggaaccga caatgcagag
     3541 atgggccatg ttgatccttt tgcttccttc ctcttctatc agagacatcc ttctcagacg
     3601 gtccacatct ttaggaaaca aggccacaat tctatctaaa tgcttctgat ttatctcata
     3661 aatgatttcc aaatgtcgag ggagcagctt ctccaccagg tccacgggcc agcgctccag
     3721 ggcttccggg agcactgtgt ggttggtgta ggcgaaggtc ttctgggtga gctcccatgc
     3781 cttggaccag ggcagttttt caatatccac aaaaatcctc atcagctcag ggatcgcgag
     3841 tgcagggtga gtgtcattca gctggatggc cacctgatcc gggaaggcat caaacacagt
     3901 tcctgcacca cgggtggagc caaacttgga ggctttgaaa cggcggatga tatcttgcaa
     3961 ggttgcagcc accacaaagt attcctgctt caatcttagc tccttccctt caaaaaaatt
     4021 gtcattggga tagaggaccc gggagatgtt ctcggccagg tttcggtcca gcacagcctg
     4081 aatgtagtct ccaacattaa agtctctgag gttaaagtca tttggtgccc gagcagacca
     4141 gaggcgcatg gtgttgacag tgttattcat gtagccgggc acgggggtgt catatggcag
     4201 agccaggacc acttgagtgt caatccactt ggtcccggtg ttggtgtgtt ctacttttcc
     4261 atagaagtgc acaggcagca tgaattctgg gcgggacttc tcccaagggt ttccatatct
     4321 gagccaatca tctgcttctt ctacctgcca tccatctcgg atcttctgat tgaaaatccc
     4381 atattcatac cgaatgccgt atccataggc tgcaagtccc agggttgcca tggaatccaa
     4441 gaagcaggca gcaagtctcc caagaccacc attgccaagt ccagcatctt cttcaatttc
     4501 ttctaactct tctatatcca atccaagctg gtaaatggcc tcatcacagg cattttgcag
     4561 accgaggttg atcatggtgt tctgtaatgt tcggcccatg taaaattcca gagagaggta
     4621 atataccctc ttggggcact tgtcgtagta gtgctgctgc gtgcggatcc agcgccccac
     4681 caggtggtcg cgcaccgtgt gcgccagcgc gaagtagtag tcgcgggtgg tggccacgtt
     4741 gcggtccttg accagcgtga agtgcaggtg ccggttgaaa ctcttcttca gctctgccac
     4801 gttctccacg cccacgatgc cgcggatgct gatctgccgc cgcttctcct ggtccgtcag
     4861 gggcttcgcc atggtgccaa cttttttgta caaagttggc attataaaaa agcattgctc
     4921 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttggggccc
     4981 gagcttaaga ctggccgtcg ttttacaacg tcgtgactgg gaaaacatcc atgctagcgt
     5041 taacgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag tcggaagact
     5101 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
     5161 cgggagcgga tttgaacgtt gtgaagcaac ggcccggagg gtggcgggca ggacgcccgc
     5221 cataaactgc caggcatcaa actaagcaga aggccatcct gacggatggc ctttttgcgt
     5281 ttctacaaac tcttcctggc tagcggtacg cgtattaatt gcgttgcgct cact
//
