LOCUS       sequence-165326-        8145 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing TSC2, part of the Target Accelerator
            Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8145)
  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 8145)
  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_TSC2_WT
FEATURES             Location/Qualifiers
     source          1..8145
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(2151..2250)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2266..2282)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2275..2297)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      2346..2432
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2524..2551
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2710..2727)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2881..3469)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2961..2980)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3562..4353)
                     /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     4784..4800
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4784..4800
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4805..4824
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4805..4823
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4841..4940
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ggactcaggc agcctgccca gaaccagctt cagcacctcc cagtcagact cctgcttcaa
       61 gcactgcagc aggacgcgga agagcaggga gtagggcacg gaccccagcc gcacggcggg
      121 gcctgcaggc gccgggccag gaggccctgt gggaggagaa agggggccgc tggtcttctt
      181 ctcagagcct ctctctggct ccatgtagtc gcagacgcag taggggctga accgcacgac
      241 tccatccttg ttgggcaggc ccaggcggtg cagtgagtcg gcccgcagca gcaacaggaa
      301 gtcaaaggcc tgcagccgga tgctgctcgc gattggcagg gtgtagctgt gcttgtagtg
      361 gagctgaatg tggctgacca gcatctcata cacacgcgtg gcgtggcttg caggcagggt
      421 gtacagcttg gtctgaagga tgaccagaag ccccaggacg gctgtcttca catcctccaa
      481 ggaggccgag tatgcggcca catccctttc ttccagctcc gggggtgggg agagggagcg
      541 ggccatcacc ttctcgatga tgtccagcag gctgttgaag tggtgtgtgt ggcagccctc
      601 tgccaggtcc accagcaact gggtggccag ctttcggacc tggtggtctt tatcctcggg
      661 gatgtgggag agctgcgaga tgaccactga gttaatcagc tcctcctcat agaactgcct
      721 gttgatgagc agcacaaagg acagcacgtc cagcaccttg atgcgcacgg cgcctcggga
      781 ctcgctcctg aagaatctct ccatcagcgc ctgcaggttc tgaatccagc cgtccttggc
      841 cgggtggatg gactgcgctc tataggagat caggttcagg agggaggact caggcctctg
      901 gtccgcacat ctctccacca gttcaaagta tctctcctga gacccgtgga actcgttctg
      961 gtcacacagc tcctccaccg tggtcaacag gtcatggacg atggtcctga gctccgggct
     1021 gtccaaggtc tggagctgct gaaggagccg ttcgatgatg ttcagcagaa tgtcccacgc
     1081 caccacctgg agctccttcc tatacttctt gatgagcctg gtgatggaca ggacgatctc
     1141 ataggacacc acctcgttcg gacatgccat ggcctggtaa aatgatggca acacagatgt
     1201 cggcgagttc ctgagagaat agagccggtg ggctccccag agagccatgc ccacaaaaaa
     1261 cacggctcct ctcagcaggg gcgcgtcctc catgtaggct ctgtcctcca tgaggtggca
     1321 catgttgtag atggcgctgt ggcccaggtg ggtgccaagg aggttccgca tcagcttcca
     1381 gcaaggctcg cagagctcct tgacgttgat ggtgcgacag agggtaacga tgaacagcgg
     1441 gaggctctca gccggcaggc agttgtagca gaccacggcg tccagcacct gcagggagac
     1501 ctctatgtcc acagaggaca cggtccggac gcacagcaga cagatcatct gaaccatcct
     1561 tgcgatgtac tcgtcgaggt aacagctatt gaatttgacc aagttcacca gcaccagaag
     1621 gaattccgag gacaagccaa catccatcca ctgcaggaca aagtcagcca gctcttcctc
     1681 caagtaggtg atgtgtctcc cattgtctgt gagggccttg aaaacctcca gcctttcgtg
     1741 aaggtcttcg ttggaagggt aatccttgat gaccttaaag aagagggctc tgaggacccc
     1801 caaacgctcg ccctgcccct gcacgatggc cttcagcaga gccagcaccg cgtgccgggc
     1861 ctccagcggc cgctccggct gcaacagatc cgcgaccgcc ttccagagtg cttccactgc
     1921 gtgctcttca aatttcttgg tttttgcgac ttcacaaatc tgccctatca tccggatgcg
     1981 attgttgagg ccacattcca tgctcagttc tctcagtatt tccgcggtga tgataaactc
     2041 cgtctgttta ccctctgcag acctgggatt tggcctcggt gttcccagtc ccaacagaat
     2101 cttaaacttc tccttcaagc ctgaatcttt gcttgttggt ttggccatgg tgccaacttt
     2161 tttgtacaaa gttggcatta taaaaaagca ttgctcatca atttgttgca acgaacaggt
     2221 cactatcagt caaaataaaa tcattatttg gggcccgagc ttaagactgg ccgtcgtttt
     2281 acaacgtcgt gactgggaaa acatccatgc tagcgttaac gcgagagtag ggaactgcca
     2341 ggcatcaaat aaaacgaaag gctcagtcgg aagactgggc ctttcgtttt atctgttgtt
     2401 tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgtga
     2461 agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaacta
     2521 agcagaaggc catcctgacg gatggccttt ttgcgtttct acaaactctt cctggctagc
     2581 ggtacgcgta ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     2641 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     2701 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2761 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2821 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2881 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2941 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3001 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3061 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3121 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3181 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3241 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3301 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3361 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3421 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3481 gatcttttct acggggtctg acgctcagtg gaacgacgcg taactcacgt taagggattt
     3541 tggtcatggg tggctcgagg gttatttgcc gactaccttg gtgatctcgc ctttcacgta
     3601 gtggacaaat tcttccaact gatctgcgcg cgaggccaag cgatcttctt cttgtccaag
     3661 ataagcctgt ctagcttcaa gtatgacggg ctgatactgg gccggcaggc gctccattgc
     3721 ccagtcggca gcgacatcct tcggcgcgat tttgccggtt actgcgctgt accaaatgcg
     3781 ggacaacgta agcactacat ttcgctcatc gccagcccag tcgggcggcg agttccatag
     3841 cgttaaggtt tcatttagcg cctcaaatag atcctgttca ggaaccggat caaagagttc
     3901 ctccgccgct ggacctacca aggcaacgct atgttctctt gcttttgtca gcaagatagc
     3961 cagatcaatg tcgatcgtgg ctggctcgaa gatacctgca agaatgtcat tgcgctgcca
     4021 ttctccaaat tgcagttcgc gcttagctgg ataacgccac ggaatgatgt cgtcgtgcac
     4081 aacaatggtg acttctacag cgcggagaat ctcgctctct ccaggggaag ccgaagtttc
     4141 caaaaggtcg ttgatcaaag ctcgccgcgt tgtttcatca agccttacgg tcaccgtaac
     4201 cagcaaatca atatcactgt gtggcttcag gccgccatcc actgcggagc cgtacaaatg
     4261 tacggccagc aacgtcggtt cgagatggcg ctcgatgacg ccaactacct ctgatagttg
     4321 agtcgatact tcggcgatca ccgcttccct cataatgttt aactttgttt tagggcgact
     4381 gccctgctgc gtaacatcgt tgctgctcca taacatcaaa catcgaccca cggcgtaacg
     4441 cgcttgctgc ttggatgccc gaggcataga ctgtacccca aaaaaacagt cataacaagc
     4501 catgaaaacc gccactgcgc cgttaccacc gctgcgttcg gtcaaggttc tggaccagtt
     4561 gcgtgagcgc atacgctact tgcattacag cttacgaacc gaacaggctt atgtccactg
     4621 ggttcgtgcc ttcatccgtt tccacggtgt gcgtcacccg gcaaccttgg gtagcagcga
     4681 agtcgaggca tttctgtcct ggctggtcta gaggaggcat gatgatatat ttttatcttg
     4741 tgcaatgtaa catcagagat tttgagacac gggccagagc tgccaggaaa cagctatgac
     4801 catgtaatac gactcactat aggggatatc agctggatgg caaataatga ttttattttg
     4861 actgatagtg acctgttcgt tgcaacaaat tgataagcaa tgctttctta taatgccaac
     4921 tttgtacaag aaagttggca acacaaactc ggtgaagtcc tccaccgagg agatgaggcg
     4981 cttccgctgg cccacctcat agccaggtgt gggctcggct ggagtctgtg caggggcttt
     5041 gctatgggac ggagggtgca ccagaggtag gctggggttg gagtaggcgg cttcctcgca
     5101 gatccgctgg cggagccgct tgatgtggcg gagccgggca atccacttgg aggggtagat
     5161 atcggtgggg ttggagcggc tatgatgcac ctgtgaggcc atatttgcgt gcagggccat
     5221 ctggcgggcc acgaagggca ggttgcggtc agacacgatc ttggccacgc tggtgtccac
     5281 aaggccctcc atgtctttcc tgcactgcag ggacaccagg ttgcactcgt agtccagcgg
     5341 ggtgaccatc acgtggacaa agttgaactg gcccttgatg gtgccaagct tgaagtcctc
     5401 accggagtca ttgtagacaa tggacacaaa gtcgttgccc aggtggcgct tcttgtcgca
     5461 gcggtgcttg tccacgtcct tggtgggcat cagggtggcg atgtggaaga cggcttgcat
     5521 gatgtcatcg tgccagcagt aggtgaactg gccgtcctca ccacacacgt ccaggcctcc
     5581 caggtacacc ttgtccggct ggcagtcctt cagctcgatg agccggccca ggcccgtcag
     5641 gaactccgtg tacctgtagg agccatgctc attggacagg atggcgagct cgctgttgct
     5701 ctggccttct ccaacataca ggacggcgat cttgtgggtg tcgtatgatg ggatctggtc
     5761 gaggagctgc accgaccgct caaaggactg tgactcattg ggcagcagga ttggcttgtt
     5821 tgactcgtcg ccaaagaagg gggaatggta gagctgcagg aacacgaaac tggggttgat
     5881 gcctggcact ttctctgcat tggaggctgt ggctctgctc tttaaggcgt ccctctctac
     5941 tctcttgccc ctgcgtgatg gggccgagtc ggagatggtg taacctcggg gccggaggcc
     6001 actgggcgag cggggggagc tggaaggcaa gggaccctcg ggctggcccc gactgtcttc
     6061 gcccgaggcc gaccaggcag cactttcccc gtccagggtc cctgactgtg accgggcctt
     6121 aacctcaggg ctcagccggc ccacgtcggc cttgtcccca gggtccccga ggatgtcctg
     6181 cagagtctgc agctcgggag aggagctgga cttgctcagg ggctgcgagg gctggaagga
     6241 gaggtccaca gagggccggc caccctccga ggagacgact cgctcgatgg ggatgcccct
     6301 gccaaccagc tcctccgcgt ggagcgactt ctcctcctgg ctggagactg aggacgacct
     6361 gctgtaggca tccgtgcgcc tgtccatgcc tagcgctgcc tcaacgtcct ccaaccctgg
     6421 gggctccacc agcacaggaa cctcgcccgg acttccctcc tccatgacca cggcggagtc
     6481 tgtgttggag cgaggcagag gagggggttt ggccgtgctg gctgccggca ccgacagtga
     6541 cttgtacagg gctgtgtccc ggtgctcctt gaagcgctca gccgccatga gggcgttaga
     6601 cagctcctgc aggggcatgt tgttgatgtc cgaggagaaa gggctgagcg ggttctccag
     6661 gctcatcagc cagctggtgt tccctgtggg cctccggacc aggatctccg cccagccctg
     6721 ggtcagcagg ggcacatagg ccgccaggtt cgtcttctcc tgcacaggaa cccgggtgcc
     6781 agctgaggcc ttctcaggtt tcgcggctgg tgcagtccgt ggtcctggag aagtggcact
     6841 gcccaggaac tgggaggccg gcacgtccag ggcgccaact cgaagaccat ggccccccga
     6901 catggaacgg acccgatccc gggccccacg ggacacctgc tgcccagcct gggactccag
     6961 cttggccggc gcctccttgg tctgtctcac atgcaccccg gggctggagc tcgactccgg
     7021 gccggactgc agctcccccg agtccaggcc tagtaacgac cgggtcccgg ttcccacgct
     7081 tgtcgtcaca gtgacaagct tgttcccaac cagccaggtt ttggtcctgc cacccgctag
     7141 gaggaactcg cccacaggag acctcttcgg gacagccgtg aagttggaga agacgtatcg
     7201 agccatcatg tccagacagg tttccgtgag ctccaggtgg aggtttttca tgctatcgtc
     7261 agcctgggcc acggagttct catctgcaga ccccaagctg gcactggtga gggacgtctg
     7321 tatcctgctg cggaccacat gttcagacac actgatgctg cggcaccgga aggcctcggc
     7381 tgcagagctc tccttgaatt ctttcacggg tggagagtta ttcaagcctt gtttgggggg
     7441 tctggctatc ctcagactct tgggtctctc gttgagacta gtactccggg ccctgaagct
     7501 gtccttctcg ggggtgtcat caaaagacaa gaggacattg gaccgcaggc ccttagtgat
     7561 gaaagggaca aaatccttcc ggaagggcag gcggcacctg atgaaccaca tggctatgac
     7621 gtgatgggcc agacacacga tgtactgatt aaacttggag gggttggtgt acggcaggga
     7681 gatggcgaac acactggcat actgctccgc ggcaaagttc ctgtagaggt gcggcagcct
     7741 ggccagagtg gacaggaact ccagcagtgg gacggccatg ctggctgtgg ctgagatgtg
     7801 cgtgagcttc accaccagaa caggcagcgc cttgatgatg atgtcaggca tctccacgct
     7861 gcagatggac aaggccacga cgcactggct ggcacagcgg tggatgaggc cctgctccag
     7921 gcagtagacc atctcgcgct gtttggtttt gtccaggtag ttatggtaag agattaatgc
     7981 tgtcagcact ggaaccacgg ccaggtgcaa gtcagttctg gagaagcctt ctggggcgcc
     8041 tcggagccgc tccagtgtct ttgggcctga aagcatggag cagagagcag agcacagctg
     8101 gtccacactg caaggggaag taaagatgag cactttatag cgcag
//
