LOCUS       sequence-165367-        6498 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing NOTCH2, part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6498)
  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 6498)
  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_NOTCH2_WT
FEATURES             Location/Qualifiers
     source          1..6498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      580..1168
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1069..1088
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1322..1339
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(1498..1525)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1617..1703)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1752..1774
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1766..1783
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1767..1783
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1799..1898
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      3636..3645
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(5607..5706)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5723..5742)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5724..5742)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5747..5763)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5747..5763)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(6194..6498,1..487)
                     /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"
ORIGIN
        1 agctaagcgc gaactgcaat ttggagaatg gcagcgcaat gacattcttg caggtatctt
       61 cgagccagcc acgatcgaca ttgatctggc tatcttgctg acaaaagcaa gagaacatag
      121 cgttgccttg gtaggtccag cggcggagga actctttgat ccggttcctg aacaggatct
      181 atttgaggcg ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggcga
      241 tgagcgaaat gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat
      301 cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc
      361 cgtcatactt gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg
      421 cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg
      481 caaataaccc tcgagccacc catgaccaaa atcccttaac gtgagttacg cgtcgttcca
      541 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
      601 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
      661 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
      721 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
      781 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
      841 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
      901 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
      961 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     1021 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     1081 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     1141 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     1201 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     1261 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     1321 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     1381 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     1441 tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa gagtttgtag aaacgcaaaa
     1501 aggccatccg tcaggatggc cttctgctta gtttgatgcc tggcagttta tggcgggcgt
     1561 cctgcccgcc accctccggg ccgttgcttc acaacgttca aatccgctcc cggcggattt
     1621 gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtcttcc
     1681 gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcgt taacgctagc
     1741 atggatgttt tcccagtcac gacgttgtaa aacgacggcc agtcttaagc tcgggcccca
     1801 aataatgatt ttattttgac tgatagtgac ctgttcgttg caacaaattg atgagcaatg
     1861 cttttttata atgccaactt tgtacaaaaa agttggcacc atgcccgccc tgcgccccgc
     1921 tctgctgtgg gcgctgctgg cgctctggct gtgctgcgcg gcccccgcgc atgcattgca
     1981 gtgtcgagat ggctatgaac cctgtgtaaa tgaaggaatg tgtgttacct accacaatgg
     2041 cacaggatac tgcaaatgtc cagaaggctt cttgggggaa tattgtcaac atcgagaccc
     2101 ctgtgagaag aaccgctgcc agaatggtgg gacttgtgtg gcccaggcca tgctggggaa
     2161 agccacgtgc cgatgtgcct cagggtttac aggagaggac tgccagtact caacatctca
     2221 tccatgcttt gtgtctcgac cctgcctgaa tggcggcaca tgccatatgc tcagccggga
     2281 tacctatgag tgcacctgtc aagtcgggtt tacaggtaag gagtgccaat ggacggatgc
     2341 ctgcctgtct catccctgtg caaatggaag tacctgtacc actgtggcca accagttctc
     2401 ctgcaaatgc ctcacaggct tcacagggca gaaatgtgag actgatgtca atgagtgtga
     2461 cattccagga cactgccagc atggtggcac ctgcctcaac ctgcctggtt cctaccagtg
     2521 ccagtgccct cagggcttca caggccagta ctgtgacagc ctgtatgtgc cctgtgcacc
     2581 ctcaccttgt gtcaatggag gcacctgtcg gcagactggt gacttcactt ttgagtgcaa
     2641 ctgccttcca ggttttgaag ggagcacctg tgagaggaat attgatgact gccctaacca
     2701 caggtgtcag aatggagggg tttgtgtgga tggggtcaac acttacaact gccgctgtcc
     2761 cccacaatgg acaggacagt tctgcacaga ggatgtggat gaatgcctgc tgcagcccaa
     2821 tgcctgtcaa aatgggggca cctgtgccaa ccgcaatgga ggctatggct gtgtatgtgt
     2881 caacggctgg agtggagatg actgcagtga gaacattgat gattgtgcct tcgcctcctg
     2941 tactccaggc tccacctgca tcgaccgtgt ggcctccttc tcttgcatgt gcccagaggg
     3001 gaaggcaggt ctcctgtgtc atctggatga tgcatgcatc agcaatcctt gccacaaggg
     3061 ggcactgtgt gacaccaacc ccctaaatgg gcaatatatt tgcacctgcc cacaaggcta
     3121 caaaggggct gactgcacag aagatgtgga tgaatgtgcc atggccaata gcaatccttg
     3181 tgagcatgca ggaaaatgtg tgaacacgga tggcgccttc cactgtgagt gtctgaaggg
     3241 ttatgcagga cctcgttgtg agatggacat caatgagtgc cattcagacc cctgccagaa
     3301 tgatgctacc tgtctggata agattggagg cttcacatgt ctgtgcatgc caggtttcaa
     3361 aggtgtgcat tgtgaattag aaataaatga atgtcagagc aacccttgtg tgaacaatgg
     3421 gcagtgtgtg gataaagtca atcgtttcca gtgcctgtgt cctcctggtt tcactgggcc
     3481 agtttgccag attgatattg atgactgttc cagtactccg tgtctgaatg gggcaaagtg
     3541 tatcgatcac ccgaatggct atgaatgcca gtgtgccaca ggtttcactg gtgtgttgtg
     3601 tgaggagaac attgacaact gtgaccccga tccttgccac catggtcagt gtcaggatgg
     3661 tattgattcc tacacctgca tctgcaatcc cgggtacatg ggcgccatct gcagtgacca
     3721 gattgatgaa tgttacagca gcccttgcct gaacgatggt cgctgcattg acctggtcaa
     3781 tggctaccag tgcaactgcc agccaggcac gtcaggggtt aattgtgaaa ttaattttga
     3841 tgactgtgca agtaaccctt gtatccatgg aatctgtatg gatggcatta atcgctacag
     3901 ttgtgtctgc tcaccaggat tcacagggca gagatgtaac attgacattg atgagtgtgc
     3961 ctccaatccc tgtcgcaagg gtgcaacatg tatcaacggt gtgaatggtt tccgctgtat
     4021 atgccccgag ggaccccatc accccagctg ctactcacag gtgaacgaat gcctgagcaa
     4081 tccctgcatc catggaaact gtactggagg tctcagtgga tataagtgtc tctgtgatgc
     4141 aggctgggtt ggcatcaact gtgaagtgga caaaaatgaa tgcctttcga atccgtgcca
     4201 gaatggagga acttgtgaca atctggtgaa tggatacagg tgtacttgca agaagggctt
     4261 taaaggctat aactgccagg tgaatattga tgaatgtgcc tcaaatccat gcctgaacca
     4321 aggaacctgc tttgatgaca taagtggcta cacttgccac tgtgtgctgc catacacagg
     4381 caagaattgt cagacagtat tggctccctg ttccccaaac ccttgtgaga atgctgctgt
     4441 ttgcaaagag tcaccaaatt ttgagagtta tacttgcttg tgtgctcctg gctggcaagg
     4501 tcagcggtgt accattgaca ttgacgagtg tatctccaag ccctgcatga accatggtct
     4561 ctgccataac acccagggca gctacatgtg tgaatgtcca ccaggcttca gtggtatgga
     4621 ctgtgaggag gacattgatg actgccttgc caatccttgc cagaatggag gttcctgtat
     4681 ggatggagtg aatactttct cctgcctctg ccttccgggt ttcactgggg ataagtacca
     4741 gacagacatg aatgagtgtc tgagtgaacc ctgtaagaat ggagggacct gctctgacta
     4801 cgtcaacagt tacacttgca agtgccaggc aggatttgat ggagtccatt gtgagaacaa
     4861 catcaatgag tgcactgaga gctcctgttt caatggtggc acatgtgttg atgggattaa
     4921 ctccttctct tgcttgtgcc ctgtgggttt cactggatcc ttctgcctcc atgagatcaa
     4981 tgaatgcagc tctcatccat gcctgaatga gggaacgtgt gttgatggcc tgggtaccta
     5041 ccgctgcagc tgccccctgg gctacactgg gaaaaactgt cagaccctgg tgaatctctg
     5101 cagtcggtct ccatgtaaaa acaaaggtac ttgcgttcag aaaaaagcag agtcccagtg
     5161 cctatgtcca tctggatggg ctggtgccta ttgtgacgtg cccaatgtct cttgtgacat
     5221 agcagcctcc aggagaggtg tgcttgttga acacttgtgc cagcactcag gtgtctgcat
     5281 caatgctggc aacacgcatt actgtcagtg ccccctgggc tatactggga gctactgtga
     5341 ggagcaactc gatgagtgtg cgtccaaccc ctgccagcac ggggcaacat gcagtgactt
     5401 cattggtgga tacagatgcg agtgtgtccc aggctatcag ggtgtcaact gtgagtatga
     5461 agtggatgag tgccagaatc agccctgcca gaatggaggc acctgtattg accttgtgaa
     5521 ccatttcaag tgctcttgcc caccaggcac tcggggtatg aaatcatcct tatccatttt
     5581 ccatccaggg cattgtctta agttattgcc aactttcttg tacaaagttg gcattataag
     5641 aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat
     5701 tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc
     5761 ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     5821 atcatcatgc ctcctctaga ccagccagga cagaaatgcc tcgacttcgc tgctacccaa
     5881 ggttgccggg tgacgcacac cgtggaaacg gatgaaggca cgaacccagt ggacataagc
     5941 ctgttcggtt cgtaagctgt aatgcaagta gcgtatgcgc tcacgcaact ggtccagaac
     6001 cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt gttatgactg
     6061 tttttttggg gtacagtcta tgcctcgggc atccaagcag caagcgcgtt acgccgtggg
     6121 tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagggcagtc gccctaaaac
     6181 aaagttaaac attatgaggg aagcggtgat cgccgaagta tcgactcaac tatcagaggt
     6241 agttggcgtc atcgagcgcc atctcgaacc gacgttgctg gccgtacatt tgtacggctc
     6301 cgcagtggat ggcggcctga agccacacag tgatattgat ttgctggtta cggtgaccgt
     6361 aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa cttcggcttc
     6421 ccctggagag agcgagattc tccgcgctgt agaagtcacc attgttgtgc acgacgacat
     6481 cattccgtgg cgttatcc
//
