LOCUS       sequence-161505-        4452 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing SMAD4, part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4452)
  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 4452)
  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_SMAD4_p.P215S
FEATURES             Location/Qualifiers
     source          1..4452
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(312..411)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(428..447)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(429..447)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(452..468)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(452..468)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             899..1690
                     /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      1783..2371
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2272..2291
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2525..2542
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2701..2728)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2820..2906)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2955..2977
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2969..2986
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2970..2986
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3002..3101
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 ggcggctact gcacaagctg cagcagctgc ccaggcagca gccgtggcag gaaacatccc
       61 tggcccagga tcagtaggtg gaatagctcc agctatcagt ctgtcagctg ctgctggaat
      121 tggtgttgat gaccttcgtc gcttatgcat actcaggatg agttttgtga aaggctgggg
      181 accggattac ccaagacaga gcatcaaaga aacaccttgc tggattgaaa ttcacttaca
      241 ccgggccctc cagctcctag acgaagtact tcataccatg ccgattgcag acccacaacc
      301 tttagactga ttgccaactt tcttgtacaa agttggcatt ataagaaagc attgcttatc
      361 aatttgttgc aacgaacagg tcactatcag tcaaaataaa atcattattt gccatccagc
      421 tgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc
      481 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgcctcct
      541 ctagaccagc caggacagaa atgcctcgac ttcgctgcta cccaaggttg ccgggtgacg
      601 cacaccgtgg aaacggatga aggcacgaac ccagtggaca taagcctgtt cggttcgtaa
      661 gctgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga ccgaacgcag
      721 cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt ttggggtaca
      781 gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt
      841 atggagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taaacattat
      901 gagggaagcg gtgatcgccg aagtatcgac tcaactatca gaggtagttg gcgtcatcga
      961 gcgccatctc gaaccgacgt tgctggccgt acatttgtac ggctccgcag tggatggcgg
     1021 cctgaagcca cacagtgata ttgatttgct ggttacggtg accgtaaggc ttgatgaaac
     1081 aacgcggcga gctttgatca acgacctttt ggaaacttcg gcttcccctg gagagagcga
     1141 gattctccgc gctgtagaag tcaccattgt tgtgcacgac gacatcattc cgtggcgtta
     1201 tccagctaag cgcgaactgc aatttggaga atggcagcgc aatgacattc ttgcaggtat
     1261 cttcgagcca gccacgatcg acattgatct ggctatcttg ctgacaaaag caagagaaca
     1321 tagcgttgcc ttggtaggtc cagcggcgga ggaactcttt gatccggttc ctgaacagga
     1381 tctatttgag gcgctaaatg aaaccttaac gctatggaac tcgccgcccg actgggctgg
     1441 cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa
     1501 aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg cccagtatca
     1561 gcccgtcata cttgaagcta gacaggctta tcttggacaa gaagaagatc gcttggcctc
     1621 gcgcgcagat cagttggaag aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt
     1681 cggcaaataa ccctcgagcc acccatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     1741 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     1801 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     1861 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     1921 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     1981 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2041 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     2101 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     2161 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     2221 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     2281 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     2341 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     2401 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     2461 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     2521 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     2581 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     2641 cagtgagcgc aacgcaatta atacgcgtac cgctagccag gaagagtttg tagaaacgca
     2701 aaaaggccat ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg
     2761 cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga
     2821 tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
     2881 tccgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct
     2941 agcatggatg ttttcccagt cacgacgttg taaaacgacg gccagtctta agctcgggcc
     3001 ccaaataatg attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgatgagca
     3061 atgctttttt ataatgccaa ctttgtacaa aaaagttggc accatggaca atatgtctat
     3121 tacgaataca ccaacaagta atgatgcctg tctgagcatt gtgcatagtt tgatgtgcca
     3181 tagacaaggt ggagagagtg aaacatttgc aaaaagagca attgaaagtt tggtaaagaa
     3241 gctgaaggag aaaaaagatg aattggattc tttaataaca gctataacta caaatggagc
     3301 tcatcctagt aaatgtgtta ccatacagag aacattggat gggaggcttc aggtggctgg
     3361 tcggaaagga tttcctcatg tgatctatgc ccgtctctgg aggtggcctg atcttcacaa
     3421 aaatgaacta aaacatgtta aatattgtca gtatgcgttt gacttaaaat gtgatagtgt
     3481 ctgtgtgaat ccatatcact acgaacgagt tgtatcacct ggaattgatc tctcaggatt
     3541 aacactgcag agtaatgctc catcaagtat gatggtgaag gatgaatatg tgcatgactt
     3601 tgagggacag ccatcgttgt ccactgaagg acattcaatt caaaccatcc agcatccacc
     3661 aagtaatcgt gcatcgacag agacatacag caccccagct ctgttagccc catctgagtc
     3721 taatgctacc agcactgcca acttttccaa cattcctgtg gcttccacaa gtcagcctgc
     3781 cagtatactg gggggcagcc atagtgaagg actgttgcag atagcatcag ggcctcagcc
     3841 aggacagcag cagaatggat ttactggtca gccagctact taccatcata acagcactac
     3901 cacctggact ggaagtagga ctgcaccata cacacctaat ttgcctcacc accaaaacgg
     3961 ccatcttcag caccacccgc ctatgccgcc ccatcccgga cattactggc ctgttcacaa
     4021 tgagcttgca ttccagcctc ccatttccaa tcatcctgct cctgagtatt ggtgttccat
     4081 tgcttacttt gaaatggatg ttcaggtagg agagacattt aaggttcctt caagctgccc
     4141 tattgttact gttgatggat acgtggaccc ttctggagga gatcgctttt gtttgggtca
     4201 actctccaat gtccacagga cagaagccat tgagagagca aggttgcaca taggcaaagg
     4261 tgtgcagttg gaatgtaaag gtgaaggtga tgtttgggtc aggtgcctta gtgaccacgc
     4321 ggtctttgta cagagttact acttagacag agaagctggg cgtgcacctg gagatgctgt
     4381 tcataagatc tacccaagtg catatataaa ggtctttgat ttgcgtcagt gtcatcgaca
     4441 gatgcagcag ca
//
