LOCUS       sequence-161682-        4614 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 4614)
  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 4614)
  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_WT
FEATURES             Location/Qualifiers
     source          1..4614
                     /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(2014..2113)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2130..2149)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2131..2149)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2154..2170)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2154..2170)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2601..3392
                     /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      3485..4073
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3974..3993
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4227..4244
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4403..4430)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4522..4608)
                     /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 catggagcca ccacggggcc cccctgccaa tggggccgag ccatcccggg
      241 cagtgggcac cgtcaaagta tacctgccca acaagcaacg cacggtggtg actgtccggg
      301 atggcatgag tgtctacgac tctctagaca aggccctgaa ggtgcggggt ctaaatcagg
      361 actgctgtgt ggtctaccga ctcatcaagg gacgaaagac ggtcactgcc tgggacacag
      421 ccattgctcc cctggatggc gaggagctca ttgtcgaggt ccttgaagat gtcccgctga
      481 ccatgcacaa ttttgtacgg aagaccttct tcagcctggc gttctgtgac ttctgcctta
      541 agtttctgtt ccatggcttc cgttgccaaa cctgtggcta caagttccac cagcattgtt
      601 cctccaaggt ccccacagtc tgtgttgaca tgagtaccaa ccgccaacag ttctaccaca
      661 gtgtccagga tttgtccgga ggctccagac agcatgaggc tccctcgaac cgccccctga
      721 atgagttgct aaccccccag ggtcccagcc cccgcaccca gcactgtgac ccggagcact
      781 tccccttccc tgccccagcc aatgcccccc tacagcgcat ccgctccacg tccactccca
      841 acgtccatat ggtcagcacc acggccccca tggactccaa cctcatccag ctcactggcc
      901 agagtttcag cactgatgct gccggtagta gaggaggtag tgatggaacc ccccggggga
      961 gccccagccc agccagcgtg tcctcgggga ggaagtcccc acattccaag tcaccagcag
     1021 agcagcgcga gcggaagtcc ttggccgatg acaagaagaa agtgaagaac ctggggtacc
     1081 gggactcagg ctattactgg gaggtaccac ccagtgaggt gcagctgctg aagaggatcg
     1141 ggacgggctc gtttggcacc gtgtttcgag ggcggtggca tggcgatgtg gccgtgaagg
     1201 tgctcaaggt gtcccagccc acagctgagc aggcccaggc tttcaagaat gagatgcagg
     1261 tgctcaggaa gacgcgacat gtcaacatct tgctgtttat gggcttcatg acccggccgg
     1321 gatttgccat catcacacag tggtgtgagg gctccagcct ctaccatcac ctgcatgtgg
     1381 ccgacacacg cttcgacatg gtccagctca tcgacgtggc ccggcagact gcccagggca
     1441 tggactacct ccatgccaag aacatcatcc accgagatct caagtctaac aacatcttcc
     1501 tacatgaggg gctcacggtg aagatcggtg actttggctt ggccacagtg aagactcgat
     1561 ggagcggggc ccagcccttg gagcagccct caggatctgt gctgtggatg gcagctgagg
     1621 tgatccgtat gcaggacccg aacccctaca gcttccagtc agacgtctat gcctacgggg
     1681 ttgtgctcta cgagcttatg actggctcac tgccttacag ccacattggc tgccgtgacc
     1741 agattatctt tatggtgggc cgtggctatc tgtccccgga cctcagcaaa atctccagca
     1801 actgccccaa ggccatgcgg cgcctgctgt ctgactgcct caagttccag cgggaggagc
     1861 ggcccctctt cccccagatc ctggccacaa ttgagctgct gcaacggtca ctccccaaga
     1921 ttgagcggag tgcctcggaa ccctccttgc accgcaccca ggccgatgag ttgcctgcct
     1981 gcctactcag cgcagcccgc cttgtgcctt gattgccaac tttcttgtac aaagttggca
     2041 ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata
     2101 aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag
     2161 ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata
     2221 aaaatatatc atcatgcctc ctctagacca gccaggacag aaatgcctcg acttcgctgc
     2281 tacccaaggt tgccgggtga cgcacaccgt ggaaacggat gaaggcacga acccagtgga
     2341 cataagcctg ttcggttcgt aagctgtaat gcaagtagcg tatgcgctca cgcaactggt
     2401 ccagaacctt gaccgaacgc agcggtggta acggcgcagt ggcggttttc atggcttgtt
     2461 atgactgttt ttttggggta cagtctatgc ctcgggcatc caagcagcaa gcgcgttacg
     2521 ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt tacgcagcag ggcagtcgcc
     2581 ctaaaacaaa gttaaacatt atgagggaag cggtgatcgc cgaagtatcg actcaactat
     2641 cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc gtacatttgt
     2701 acggctccgc agtggatggc ggcctgaagc cacacagtga tattgatttg ctggttacgg
     2761 tgaccgtaag gcttgatgaa acaacgcggc gagctttgat caacgacctt ttggaaactt
     2821 cggcttcccc tggagagagc gagattctcc gcgctgtaga agtcaccatt gttgtgcacg
     2881 acgacatcat tccgtggcgt tatccagcta agcgcgaact gcaatttgga gaatggcagc
     2941 gcaatgacat tcttgcaggt atcttcgagc cagccacgat cgacattgat ctggctatct
     3001 tgctgacaaa agcaagagaa catagcgttg ccttggtagg tccagcggcg gaggaactct
     3061 ttgatccggt tcctgaacag gatctatttg aggcgctaaa tgaaacctta acgctatgga
     3121 actcgccgcc cgactgggct ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt
     3181 ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg
     3241 agcgcctgcc ggcccagtat cagcccgtca tacttgaagc tagacaggct tatcttggac
     3301 aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc cactacgtga
     3361 aaggcgagat caccaaggta gtcggcaaat aaccctcgag ccacccatga ccaaaatccc
     3421 ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3481 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3541 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3601 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     3661 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3721 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3781 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3841 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3901 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     3961 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4021 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4081 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     4141 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     4201 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     4261 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     4321 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt accgctagcc
     4381 aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttagtttg
     4441 atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac
     4501 gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
     4561 gataaaacga aaggcccagt cttccgactg agcctttcgt tttatttgat gcct
//
