LOCUS       sequence-168678-        4197 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing SMAD2 ,part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4197)
  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 4197)
  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_SMAD2_WT
FEATURES             Location/Qualifiers
     source          1..4197
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     43..65
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     57..74
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     58..74
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    90..189
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(1597..1696)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1713..1732)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1714..1732)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1737..1753)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1737..1753)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2184..2975
                     /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      3068..3656
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3557..3576
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3810..3827
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3986..4013)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4105..4191)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta
       61 aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga
      121 cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa
      181 aagttggcac catgtcgtcc atcttgccat tcacgccgcc agttgtgaag agactgctgg
      241 gatggaagaa gtcagctggt gggtctggag gagcaggcgg aggagagcag aatgggcagg
      301 aagaaaagtg gtgtgagaaa gcagtgaaaa gtctggtgaa gaagctaaag aaaacaggac
      361 gattagatga gcttgagaaa gccatcacca ctcaaaactg taatactaaa tgtgttacca
      421 taccaagcac ttgctctgaa atttggggac tgagtacacc aaatacgata gatcagtggg
      481 atacaacagg cctttacagc ttctctgaac aaaccaggtc tcttgatggt cgtctccagg
      541 tatcccatcg aaaaggattg ccacatgtta tatattgccg attatggcgc tggcctgatc
      601 ttcacagtca tcatgaactc aaggcaattg aaaactgcga atatgctttt aatcttaaaa
      661 aggatgaagt atgtgtaaac ccttaccact atcagagagt tgagacacca gttttgcctc
      721 cagtattagt gccccgacac accgagatcc taacagaact tccgcctctg gatgactata
      781 ctcactccat tccagaaaac actaacttcc cagcaggaat tgagccacag agtaattata
      841 ttccagaaac gccacctcct ggatatatca gtgaagatgg agaaacaagt gaccaacagt
      901 tgaatcaaag tatggacaca ggctctccag cagaactatc tcctactact ctttcccctg
      961 ttaatcatag cttggattta cagccagtta cttactcaga acctgcattt tggtgttcga
     1021 tagcatatta tgaattaaat cagagggttg gagaaacctt ccatgcatca cagccctcac
     1081 tcactgtaga tggctttaca gacccatcaa attcagagag gttctgctta ggtttactct
     1141 ccaatgttaa ccgaaatgcc acggtagaaa tgacaagaag gcatatagga agaggagtgc
     1201 gcttatacta cataggtggg gaagtttttg ctgagtgcct aagtgatagt gcaatctttg
     1261 tgcagagccc caattgtaat cagagatatg gctggcaccc tgcaacagtg tgtaaaattc
     1321 caccaggctg taatctgaag atcttcaaca accaggaatt tgctgctctt ctggctcagt
     1381 ctgttaatca gggttttgaa gccgtctatc agctaactag aatgtgcacc ataagaatga
     1441 gttttgtgaa agggtgggga gcagaatacc gaaggcagac ggtaacaagt actccttgct
     1501 ggattgaact tcatctgaat ggacctctac agtggttgga caaagtatta actcagatgg
     1561 gatccccttc agtgcgttgc tcaagcatgt catgattgcc aactttcttg tacaaagttg
     1621 gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa
     1681 ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca
     1741 tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag
     1801 ataaaaatat atcatcatgc ctcctctaga ccagccagga cagaaatgcc tcgacttcgc
     1861 tgctacccaa ggttgccggg tgacgcacac cgtggaaacg gatgaaggca cgaacccagt
     1921 ggacataagc ctgttcggtt cgtaagctgt aatgcaagta gcgtatgcgc tcacgcaact
     1981 ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt
     2041 gttatgactg tttttttggg gtacagtcta tgcctcgggc atccaagcag caagcgcgtt
     2101 acgccgtggg tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagggcagtc
     2161 gccctaaaac aaagttaaac attatgaggg aagcggtgat cgccgaagta tcgactcaac
     2221 tatcagaggt agttggcgtc atcgagcgcc atctcgaacc gacgttgctg gccgtacatt
     2281 tgtacggctc cgcagtggat ggcggcctga agccacacag tgatattgat ttgctggtta
     2341 cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa
     2401 cttcggcttc ccctggagag agcgagattc tccgcgctgt agaagtcacc attgttgtgc
     2461 acgacgacat cattccgtgg cgttatccag ctaagcgcga actgcaattt ggagaatggc
     2521 agcgcaatga cattcttgca ggtatcttcg agccagccac gatcgacatt gatctggcta
     2581 tcttgctgac aaaagcaaga gaacatagcg ttgccttggt aggtccagcg gcggaggaac
     2641 tctttgatcc ggttcctgaa caggatctat ttgaggcgct aaatgaaacc ttaacgctat
     2701 ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca
     2761 tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa
     2821 tggagcgcct gccggcccag tatcagcccg tcatacttga agctagacag gcttatcttg
     2881 gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gtccactacg
     2941 tgaaaggcga gatcaccaag gtagtcggca aataaccctc gagccaccca tgaccaaaat
     3001 cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3061 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3121 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3181 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     3241 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     3301 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3361 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3421 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     3481 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     3541 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     3601 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     3661 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     3721 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     3781 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     3841 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     3901 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta
     3961 gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttagt
     4021 ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac
     4081 aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca
     4141 acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt gatgcct
//
