LOCUS       sequence-161695-        4609 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing ARAF, part of the Target Accelerator
            Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4609)
  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 4609)
  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_ARAF_p.S214F_p.D429A
FEATURES             Location/Qualifiers
     source          1..4609
                     /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(2009..2108)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2125..2144)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2126..2144)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2149..2165)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2149..2165)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2596..3387
                     /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      3480..4068
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3969..3988
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4222..4239
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4398..4425)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4517..4603)
                     /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 ggagccacca cggggccccc ctgccaatgg ggccgagcca tcccgggcag
      241 tgggcaccgt caaagtatac ctgcccaaca agcaacgcac ggtggtgact gtccgggatg
      301 gcatgagtgt ctacgactct ctagacaagg ccctgaaggt gcggggtcta aatcaggact
      361 gctgtgtggt ctaccgactc atcaagggac gaaagacggt cactgcctgg gacacagcca
      421 ttgctcccct ggatggcgag gagctcattg tcgaggtcct tgaagatgtc ccgctgacca
      481 tgcacaattt tgtacggaag accttcttca gcctggcgtt ctgtgacttc tgccttaagt
      541 ttctgttcca tggcttccgt tgccaaacct gtggctacaa gttccaccag cattgttcct
      601 ccaaggtccc cacagtctgt gttgacatga gtaccaaccg ccaacagttc taccacagtg
      661 tccaggattt gtccggaggc tccagacagc atgaggctcc ctcgaaccgc cccctgaatg
      721 agttgctaac cccccagggt cccagccccc gcacccagca ctgtgacccg gagcacttcc
      781 ccttccctgc cccagccaat gcccccctac agcgcatccg ctccacgttc actcccaacg
      841 tccatatggt cagcaccacg gcccccatgg actccaacct catccagctc actggccaga
      901 gtttcagcac tgatgctgcc ggtagtagag gaggtagtga tggaaccccc cgggggagcc
      961 ccagcccagc cagcgtgtcc tcggggagga agtccccaca ttccaagtca ccagcagagc
     1021 agcgcgagcg gaagtccttg gccgatgaca agaagaaagt gaagaacctg gggtaccggg
     1081 actcaggcta ttactgggag gtaccaccca gtgaggtgca gctgctgaag aggatcggga
     1141 cgggctcgtt tggcaccgtg tttcgagggc ggtggcatgg cgatgtggcc gtgaaggtgc
     1201 tcaaggtgtc ccagcccaca gctgagcagg cccaggcttt caagaatgag atgcaggtgc
     1261 tcaggaagac gcgacatgtc aacatcttgc tgtttatggg cttcatgacc cggccgggat
     1321 ttgccatcat cacacagtgg tgtgagggct ccagcctcta ccatcacctg catgtggccg
     1381 acacacgctt cgacatggtc cagctcatcg acgtggcccg gcagactgcc cagggcatgg
     1441 actacctcca tgccaagaac atcatccacc gagctctcaa gtctaacaac atcttcctac
     1501 atgaggggct cacggtgaag atcggtgact ttggcttggc cacagtgaag actcgatgga
     1561 gcggggccca gcccttggag cagccctcag gatctgtgct gtggatggca gctgaggtga
     1621 tccgtatgca ggacccgaac ccctacagct tccagtcaga cgtctatgcc tacggggttg
     1681 tgctctacga gcttatgact ggctcactgc cttacagcca cattggctgc cgtgaccaga
     1741 ttatctttat ggtgggccgt ggctatctgt ccccggacct cagcaaaatc tccagcaact
     1801 gccccaaggc catgcggcgc ctgctgtctg actgcctcaa gttccagcgg gaggagcggc
     1861 ccctcttccc ccagatcctg gccacaattg agctgctgca acggtcactc cccaagattg
     1921 agcggagtgc ctcggaaccc tccttgcacc gcacccaggc cgatgagttg cctgcctgcc
     1981 tactcagcgc agcccgcctt gtgccttagc ccaactttct tgtacaaagt tggcattata
     2041 agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc
     2101 attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt catagctgtt
     2161 tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat
     2221 atatcatcat gcctcctcta gaccagccag gacagaaatg cctcgacttc gctgctaccc
     2281 aaggttgccg ggtgacgcac accgtggaaa cggatgaagg cacgaaccca gtggacataa
     2341 gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga
     2401 accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac
     2461 tgtttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg
     2521 ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag tcgccctaaa
     2581 acaaagttaa acattatgag ggaagcggtg atcgccgaag tatcgactca actatcagag
     2641 gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc
     2701 tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt tacggtgacc
     2761 gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga aacttcggct
     2821 tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt gcacgacgac
     2881 atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg gcagcgcaat
     2941 gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc tatcttgctg
     3001 acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga actctttgat
     3061 ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct atggaactcg
     3121 ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg catttggtac
     3181 agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc
     3241 ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct tggacaagaa
     3301 gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc
     3361 gagatcacca aggtagtcgg caaataaccc tcgagccacc catgaccaaa atcccttaac
     3421 gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     3481 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3541 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3601 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     3661 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     3721 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3781 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3841 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     3901 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     3961 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4021 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4081 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     4141 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     4201 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     4261 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     4321 cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa
     4381 gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc
     4441 tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca
     4501 aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa
     4561 aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcct
//
