LOCUS       sequence-167352-        4194 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 4194)
  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 4194)
  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_p.D304G
FEATURES             Location/Qualifiers
     source          1..4194
                     /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(1594..1693)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1710..1729)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1711..1729)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1734..1750)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1734..1750)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2181..2972
                     /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      3065..3653
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3554..3573
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3807..3824
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3983..4010)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4102..4188)
                     /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 aagttggcat gtcgtccatc ttgccattca cgccgccagt tgtgaagaga ctgctgggat
      241 ggaagaagtc agctggtggg tctggaggag caggcggagg agagcagaat gggcaggaag
      301 aaaagtggtg tgagaaagca gtgaaaagtc tggtgaagaa gctaaagaaa acaggacgat
      361 tagatgagct tgagaaagcc atcaccactc aaaactgtaa tactaaatgt gttaccatac
      421 caagcacttg ctctgaaatt tggggactga gtacaccaaa tacgatagat cagtgggata
      481 caacaggcct ttacagcttc tctgaacaaa ccaggtctct tgatggtcgt ctccaggtat
      541 cccatcgaaa aggattgcca catgttatat attgccgatt atggcgctgg cctgatcttc
      601 acagtcatca tgaactcaag gcaattgaaa actgcgaata tgcttttaat cttaaaaagg
      661 atgaagtatg tgtaaaccct taccactatc agagagttga gacaccagtt ttgcctccag
      721 tattagtgcc ccgacacacc gagatcctaa cagaacttcc gcctctggat gactatactc
      781 actccattcc agaaaacact aacttcccag caggaattga gccacagagt aattatattc
      841 cagaaacgcc acctcctgga tatatcagtg aagatggaga aacaagtgac caacagttga
      901 atcaaagtat ggacacaggc tctccagcag aactatctcc tactactctt tcccctgtta
      961 atcatagctt ggatttacag ccagttactt actcagaacc tgcattttgg tgttcgatag
     1021 catattatga attaaatcag agggttggag aaaccttcca tgcatcacag ccctcactca
     1081 ctgtagatgg ctttacaggc ccatcaaatt cagagaggtt ctgcttaggt ttactctcca
     1141 atgttaaccg aaatgccacg gtagaaatga caagaaggca tataggaaga ggagtgcgct
     1201 tatactacat aggtggggaa gtttttgctg agtgcctaag tgatagtgca atctttgtgc
     1261 agagccccaa ttgtaatcag agatatggct ggcaccctgc aacagtgtgt aaaattccac
     1321 caggctgtaa tctgaagatc ttcaacaacc aggaatttgc tgctcttctg gctcagtctg
     1381 ttaatcaggg ttttgaagcc gtctatcagc taactagaat gtgcaccata agaatgagtt
     1441 ttgtgaaagg gtggggagca gaataccgaa ggcagacggt aacaagtact ccttgctgga
     1501 ttgaacttca tctgaatgga cctctacagt ggttggacaa agtattaact cagatgggat
     1561 ccccttcagt gcgttgctca agcatgtcat gattgccaac tttcttgtac aaagttggca
     1621 ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata
     1681 aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag
     1741 ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata
     1801 aaaatatatc atcatgcctc ctctagacca gccaggacag aaatgcctcg acttcgctgc
     1861 tacccaaggt tgccgggtga cgcacaccgt ggaaacggat gaaggcacga acccagtgga
     1921 cataagcctg ttcggttcgt aagctgtaat gcaagtagcg tatgcgctca cgcaactggt
     1981 ccagaacctt gaccgaacgc agcggtggta acggcgcagt ggcggttttc atggcttgtt
     2041 atgactgttt ttttggggta cagtctatgc ctcgggcatc caagcagcaa gcgcgttacg
     2101 ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt tacgcagcag ggcagtcgcc
     2161 ctaaaacaaa gttaaacatt atgagggaag cggtgatcgc cgaagtatcg actcaactat
     2221 cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc gtacatttgt
     2281 acggctccgc agtggatggc ggcctgaagc cacacagtga tattgatttg ctggttacgg
     2341 tgaccgtaag gcttgatgaa acaacgcggc gagctttgat caacgacctt ttggaaactt
     2401 cggcttcccc tggagagagc gagattctcc gcgctgtaga agtcaccatt gttgtgcacg
     2461 acgacatcat tccgtggcgt tatccagcta agcgcgaact gcaatttgga gaatggcagc
     2521 gcaatgacat tcttgcaggt atcttcgagc cagccacgat cgacattgat ctggctatct
     2581 tgctgacaaa agcaagagaa catagcgttg ccttggtagg tccagcggcg gaggaactct
     2641 ttgatccggt tcctgaacag gatctatttg aggcgctaaa tgaaacctta acgctatgga
     2701 actcgccgcc cgactgggct ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt
     2761 ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg
     2821 agcgcctgcc ggcccagtat cagcccgtca tacttgaagc tagacaggct tatcttggac
     2881 aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc cactacgtga
     2941 aaggcgagat caccaaggta gtcggcaaat aaccctcgag ccacccatga ccaaaatccc
     3001 ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3061 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3121 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3181 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     3241 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3301 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3361 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3421 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3481 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     3541 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     3601 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     3661 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     3721 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     3781 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     3841 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     3901 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt accgctagcc
     3961 aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttagtttg
     4021 atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac
     4081 gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
     4141 gataaaacga aaggcccagt cttccgactg agcctttcgt tttatttgat gcct
//
