LOCUS       sequence-167103-        5937 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing RASA1 , part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5937)
  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 5937)
  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_RASA1_p.E510K
FEATURES             Location/Qualifiers
     source          1..5937
                     /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(3337..3436)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3453..3472)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3454..3472)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3477..3493)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3477..3493)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3924..4715
                     /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      4808..5396
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5297..5316
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5550..5567
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(5726..5753)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5845..5931)
                     /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 catgatggcg gccgaggccg gcagtgagga gggcggcccg gtaacagccg
      241 gagctggagg aggcggcgcg gcagcgggct ccagtgccta tcccgcagtg tgtcgggtga
      301 agatacccgc ggccctgcct gtggcagccg ccccctatcc tgggctggtg gagaccggag
      361 tggctggaac tctgggtggc ggagccgctt tggggtcaga gttcctagga gccgggtctg
      421 tggcaggggc actgggggga gctggactga cagggggagg tactgctgct ggcgtagctg
      481 gtgctgctgc tggcgtggcc ggtgctgctg ttgctggacc tagtggagac atggctctca
      541 ccaaactgcc cacttcgttg cttgctgaga ctctcgggcc aggcggcggt tttccccctc
      601 tgccccctcc cccttacctg ccccctttgg gggcgggcct cgggacagtg gacgaaggtg
      661 actctctgga tggaccagaa tacgaggagg aagaggtggc cataccgttg accgctcctc
      721 caactaacca gtggtatcac ggaaaacttg acagaacgat agcagaagaa cgcctcaggc
      781 aggcagggaa gtctggcagt tatcttataa gagagagtga tcggaggcca gggtcctttg
      841 tactttcatt tcttagccag atgaatgttg tcaaccattt taggattatt gctatgtgtg
      901 gagattacta cattggtgga agacgttttt cttcactgtc agacctaata ggttattaca
      961 gtcatgtttc ttgtttgctt aaaggagaaa aattacttta cccagttgca ccaccagagc
     1021 cagtagaaga tagaaggcgt gtacgagcta ttctacctta cacaaaagta ccagacactg
     1081 atgaaataag tttcttaaaa ggagatatgt tcattgttca taatgaatta gaagatggat
     1141 ggatgtgggt tacaaattta agaacagatg aacaaggcct tattgttgaa gacctagtag
     1201 aagaggtggg ccgggaagaa gatccacatg aaggaaaaat atggttccat gggaagattt
     1261 ccaaacagga agcttataat ttactaatga cagttggtca agtctgcagt tttcttgtga
     1321 ggccctcaga taatactcct ggcgattatt cactttattt ccggaccaat gaaaatattc
     1381 agcgatttaa aatatgtcca acgccaaaca atcagtttat gatgggaggc cggtattata
     1441 acagcattgg ggacatcata gatcactatc gaaaagaaca gattgttgaa ggatattatc
     1501 ttaaggaacc tgtaccaatg caggatcaag aacaagtact caatgacaca gtggatggca
     1561 aggaaatcta taataccatc cgtcgtaaaa caaaggatgc cttttataaa aacattgtta
     1621 agaaaggtta tcttctgaaa aagggcaaag gaaaacgttg gaaaaattta tattttatct
     1681 tagagggtag tgatgcccaa cttatttatt ttgaaagcaa aaaacgagct accaaaccaa
     1741 aaggattaat agatctcagt gtatgttctg tctatgtcgt tcatgatagt ctctttggca
     1801 ggccaaactg ttttcagata gtagttcagc actttagtga agaacattac atcttttact
     1861 ttgcaggaga aactccagaa caagcagagg attggatgaa aggtctgcag gcattttgca
     1921 atttacggaa aagtagtcca gggacatcca ataaacgcct tcgtcaggtc agcagccttg
     1981 ttttacatat tgaagaagcc cataaactcc cagtaaaaca ttttactaat ccatattgta
     2041 acatctacct gaatagtgtc caagtagcaa aaactcatgc aagggaaggg caaaacccag
     2101 tatggtcaga agagtttgtc tttgatgatc ttcctcctga catcaataga tttgaaataa
     2161 ctcttagtaa taaaacaaag aaaagcaaag atcctgatat cttatttatg cgctgccagt
     2221 tgagccgatt acagaaaggg catgccacag atgaatggtt tctgctcagc tcccatatac
     2281 cattaaaagg tattgaacca gggtccctgc gtgttcgagc acgatactct atggaaaaaa
     2341 tcatgccaga agaagagtac agtgaattta aagagcttat actgcaaaag gaacttcatg
     2401 tagtctatgc tttatcacat gtatgtggac aagaccgaac actactggcc agcatcctac
     2461 tgaggatttt tcttcacgaa aagcttgaat cgttgttgtt atgcacacta aatgacagag
     2521 aaataagcat ggaagatgaa gccactaccc tatttcgagc cacaacactt gcaagcacct
     2581 tgatggagca gtatatgaaa gccactgcta cacagtttgt tcatcatgct ttgaaagact
     2641 ctattttaaa gataatggaa agcaagcagt cttgtgagtt aagtccatca aagttagaaa
     2701 aaaatgaaga tgtgaacact aatttaacac acctattgaa catactttca gagcttgtgg
     2761 agaaaatatt catggcttca gaaatacttc caccgacatt gagatatatt tatgggtgtt
     2821 tacagaaatc tgttcagcat aagtggccta caaataccac catgagaaca agagttgtta
     2881 gtggttttgt ttttcttcga ctcatctgtc ctgccatcct gaatccacgg atgttcaata
     2941 tcatctcaga ttctccatct cctattgctg caagaacact gatattagtg gctaaatctg
     3001 tgcagaactt agcaaatctt gtggaatttg gagctaagga gccctacatg gaaggtgtca
     3061 atccattcat caaaagcaac aaacatcgta tgatcatgtt tttagatgaa cttgggaatg
     3121 tacctgaact tccggacact acagagcatt ctagaacgga cctgtcccgt gatttagcag
     3181 cattgcatga gatttgcgtg gctcattcag atgaacttcg aacgctcagt aatgagcgtg
     3241 gtgcacagca gcacgtattg aaaaagcttc tggctataac agaactgctt caacaaaaac
     3301 aaaaccagta tacaaaaacc aatgatgtca ggtgattgcc aactttcttg tacaaagttg
     3361 gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa
     3421 ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca
     3481 tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag
     3541 ataaaaatat atcatcatgc ctcctctaga ccagccagga cagaaatgcc tcgacttcgc
     3601 tgctacccaa ggttgccggg tgacgcacac cgtggaaacg gatgaaggca cgaacccagt
     3661 ggacataagc ctgttcggtt cgtaagctgt aatgcaagta gcgtatgcgc tcacgcaact
     3721 ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt
     3781 gttatgactg tttttttggg gtacagtcta tgcctcgggc atccaagcag caagcgcgtt
     3841 acgccgtggg tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagggcagtc
     3901 gccctaaaac aaagttaaac attatgaggg aagcggtgat cgccgaagta tcgactcaac
     3961 tatcagaggt agttggcgtc atcgagcgcc atctcgaacc gacgttgctg gccgtacatt
     4021 tgtacggctc cgcagtggat ggcggcctga agccacacag tgatattgat ttgctggtta
     4081 cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa
     4141 cttcggcttc ccctggagag agcgagattc tccgcgctgt agaagtcacc attgttgtgc
     4201 acgacgacat cattccgtgg cgttatccag ctaagcgcga actgcaattt ggagaatggc
     4261 agcgcaatga cattcttgca ggtatcttcg agccagccac gatcgacatt gatctggcta
     4321 tcttgctgac aaaagcaaga gaacatagcg ttgccttggt aggtccagcg gcggaggaac
     4381 tctttgatcc ggttcctgaa caggatctat ttgaggcgct aaatgaaacc ttaacgctat
     4441 ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca
     4501 tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa
     4561 tggagcgcct gccggcccag tatcagcccg tcatacttga agctagacag gcttatcttg
     4621 gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gtccactacg
     4681 tgaaaggcga gatcaccaag gtagtcggca aataaccctc gagccaccca tgaccaaaat
     4741 cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4801 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4861 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4921 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4981 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5041 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5101 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5161 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5221 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5281 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5341 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5401 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     5461 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     5521 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     5581 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     5641 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta
     5701 gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttagt
     5761 ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac
     5821 aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca
     5881 acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt gatgcct
//
