LOCUS       sequence-166932-        4503 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing ARSI ,part of the Target Accelerator
            Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4503)
  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 4503)
  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_ARSI_p.P228L
FEATURES             Location/Qualifiers
     source          1..4503
                     /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(1903..2002)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2019..2038)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2020..2038)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2043..2059)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2043..2059)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2490..3281
                     /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      3374..3962
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3863..3882
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4116..4133
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4292..4319)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4411..4497)
                     /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 catgcacacc ctcactggct tctccctggt cagcctgctc agcttcggct
      241 acctgtcctg ggactgggcc aagccgagct tcgtggccga cgggcccggg gaggctggcg
      301 agcagccctc ggccgctccg ccccagcctc cccacatcat cttcatcctc acggacgacc
      361 aaggctacca cgacgtgggc taccatggtt cagatatcga gacccctacg ctggacaggc
      421 tggcggccaa gggggtcaag ttggagaatt attacatcca gcccatctgc acgccttcgc
      481 ggagccagct cctcactggc aggtaccaga tccacacagg actccagcat tccatcatcc
      541 gcccacagca gcccaactgc ctgcccctgg accaggtgac actgccacag aagctgcagg
      601 aggcaggtta ttccacccat atggtgggca agtggcacct gggcttctac cggaaggagt
      661 gtctgcccac ccgtcggggc ttcgacacct tcctgggctc gctcacgggc aatgtggact
      721 attacaccta tgacaactgt gatggcccag gcgtgtgcgg cttcgacctg cacgagggtg
      781 agaatgtggc ctgggggctc agcggccagt actccactat gctttacgcc cagcgcgcca
      841 gccatatcct ggccagccac agccctcagc gtctcctctt cctctatgtg gccttccagg
      901 cagtacacac acccctgcag tcccctcgtg agtacctgta ccgctaccgc accatgggca
      961 atgtggcccg gcggaagtac gcggccatgg tgacctgcat ggatgaggct gtgcgcaaca
     1021 tcacctgggc cctcaagcgc tacggtttct acaacaacag tgtcatcatc ttctccagtg
     1081 acaatggtgg ccagactttc tcggggggca gcaactggcc gctccgagga cgcaagggca
     1141 cttattggga aggtggcgtg cggggcctag gctttgtcca cagtcccctg ctcaagcgaa
     1201 agcaacggac aagccgggca ctgatgcaca tcactgactg gtacccgacc ctggtgggtc
     1261 tggcaggtgg taccacctca gcagccgatg ggctagatgg ctacgacgtg tggccggcca
     1321 tcagcgaggg ccgggcctca ccacgcacgg agatcctgca caacattgac ccactctaca
     1381 accatgccca gcatggctcc ctggagggcg gctttggcat ctggaacacc gccgtgcagg
     1441 ctgccatccg cgtgggtgag tggaagctgc tgacaggaga ccccggctat ggcgattgga
     1501 tcccaccgca gacactggcc accttcccgg gtagctggtg gaacctggaa cgaatggcca
     1561 gtgtccgcca ggccgtgtgg ctcttcaaca tcagtgctga cccttatgaa cgggaggacc
     1621 tggctggcca gcggcctgat gtggtccgca ccctgctggc tcgcctggcc gaatataacc
     1681 gcacagccat cccggtacgc tacccagctg agaacccccg ggctcatcct gactttaatg
     1741 ggggtgcttg ggggccctgg gccagtgatg aggaagagga ggaagaggaa gggagggctc
     1801 gaagcttctc ccggggtcgt cgcaagaaaa aatgcaagat ttgcaagctt cgatcctttt
     1861 tccgtaaact caacaccagg ctaatgtccc aacggatctg attgccaact ttcttgtaca
     1921 aagttggcat tataagaaag cattgcttat caatttgttg caacgaacag gtcactatca
     1981 gtcaaaataa aatcattatt tgccatccag ctgatatccc ctatagtgag tcgtattaca
     2041 tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg
     2101 cacaagataa aaatatatca tcatgcctcc tctagaccag ccaggacaga aatgcctcga
     2161 cttcgctgct acccaaggtt gccgggtgac gcacaccgtg gaaacggatg aaggcacgaa
     2221 cccagtggac ataagcctgt tcggttcgta agctgtaatg caagtagcgt atgcgctcac
     2281 gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca
     2341 tggcttgtta tgactgtttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag
     2401 cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg
     2461 gcagtcgccc taaaacaaag ttaaacatta tgagggaagc ggtgatcgcc gaagtatcga
     2521 ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg
     2581 tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc
     2641 tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt
     2701 tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg
     2761 ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag
     2821 aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc
     2881 tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg
     2941 aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa
     3001 cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt
     3061 cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact
     3121 gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt
     3181 atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc
     3241 actacgtgaa aggcgagatc accaaggtag tcggcaaata accctcgagc cacccatgac
     3301 caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga
     3361 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     3421 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3481 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     3541 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3601 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3661 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3721 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     3781 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3841 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3901 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3961 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     4021 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     4081 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     4141 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     4201 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta
     4261 ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg
     4321 cttagtttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg
     4381 cttcacaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag cgttcaccga
     4441 caaacaacag ataaaacgaa aggcccagtc ttccgactga gcctttcgtt ttatttgatg
     4501 cct
//
