LOCUS       sequence-168905-        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_WT
FEATURES             Location/Qualifiers
     source          1..5937
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3816..3915)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3932..3951)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3933..3951)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3956..3972)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3956..3972)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4403..5194
                     /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      5287..5875
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5776..5795
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agttggcacc atgatggcgg ccgaggccgg cagtgaggag ggcggcccgg taacagccgg
      721 agctggagga ggcggcgcgg cagcgggctc cagtgcctat cccgcagtgt gtcgggtgaa
      781 gatacccgcg gccctgcctg tggcagccgc cccctatcct gggctggtgg agaccggagt
      841 ggctggaact ctgggtggcg gagccgcttt ggggtcagag ttcctaggag ccgggtctgt
      901 ggcaggggca ctggggggag ctggactgac agggggaggt actgctgctg gcgtagctgg
      961 tgctgctgct ggcgtggccg gtgctgctgt tgctggacct agtggagaca tggctctcac
     1021 caaactgccc acttcgttgc ttgctgagac tctcgggcca ggcggcggtt ttccccctct
     1081 gccccctccc ccttacctgc cccctttggg ggcgggcctc gggacagtgg acgaaggtga
     1141 ctctctggat ggaccagaat acgaggagga agaggtggcc ataccgttga ccgctcctcc
     1201 aactaaccag tggtatcacg gaaaacttga cagaacgata gcagaagaac gcctcaggca
     1261 ggcagggaag tctggcagtt atcttataag agagagtgat cggaggccag ggtcctttgt
     1321 actttcattt cttagccaga tgaatgttgt caaccatttt aggattattg ctatgtgtgg
     1381 agattactac attggtggaa gacgtttttc ttcactgtca gacctaatag gttattacag
     1441 tcatgtttct tgtttgctta aaggagaaaa attactttac ccagttgcac caccagagcc
     1501 agtagaagat agaaggcgtg tacgagctat tctaccttac acaaaagtac cagacactga
     1561 tgaaataagt ttcttaaaag gagatatgtt cattgttcat aatgaattag aagatggatg
     1621 gatgtgggtt acaaatttaa gaacagatga acaaggcctt attgttgaag acctagtaga
     1681 agaggtgggc cgggaagaag atccacatga aggaaaaata tggttccatg ggaagatttc
     1741 caaacaggaa gcttataatt tactaatgac agttggtcaa gtctgcagtt ttcttgtgag
     1801 gccctcagat aatactcctg gcgattattc actttatttc cggaccaatg aaaatattca
     1861 gcgatttaaa atatgtccaa cgccaaacaa tcagtttatg atgggaggcc ggtattataa
     1921 cagcattggg gacatcatag atcactatcg aaaagaacag attgttgaag gatattatct
     1981 taaggaacct gtaccaatgc aggatcaaga acaagtactc aatgacacag tggatggcaa
     2041 ggaaatctat aataccatcc gtcgtaaaac aaaggatgcc ttttataaaa acattgttaa
     2101 gaaaggttat cttctgaaaa agggcaaagg aaaacgttgg aaaaatttat attttatctt
     2161 agagggtagt gatgcccaac ttatttattt tgaaagcgaa aaacgagcta ccaaaccaaa
     2221 aggattaata gatctcagtg tatgttctgt ctatgtcgtt catgatagtc tctttggcag
     2281 gccaaactgt tttcagatag tagttcagca ctttagtgaa gaacattaca tcttttactt
     2341 tgcaggagaa actccagaac aagcagagga ttggatgaaa ggtctgcagg cattttgcaa
     2401 tttacggaaa agtagtccag ggacatccaa taaacgcctt cgtcaggtca gcagccttgt
     2461 tttacatatt gaagaagccc ataaactccc agtaaaacat tttactaatc catattgtaa
     2521 catctacctg aatagtgtcc aagtagcaaa aactcatgca agggaagggc aaaacccagt
     2581 atggtcagaa gagtttgtct ttgatgatct tcctcctgac atcaatagat ttgaaataac
     2641 tcttagtaat aaaacaaaga aaagcaaaga tcctgatatc ttatttatgc gctgccagtt
     2701 gagccgatta cagaaagggc atgccacaga tgaatggttt ctgctcagct cccatatacc
     2761 attaaaaggt attgaaccag ggtccctgcg tgttcgagca cgatactcta tggaaaaaat
     2821 catgccagaa gaagagtaca gtgaatttaa agagcttata ctgcaaaagg aacttcatgt
     2881 agtctatgct ttatcacatg tatgtggaca agaccgaaca ctactggcca gcatcctact
     2941 gaggattttt cttcacgaaa agcttgaatc gttgttgtta tgcacactaa atgacagaga
     3001 aataagcatg gaagatgaag ccactaccct atttcgagcc acaacacttg caagcacctt
     3061 gatggagcag tatatgaaag ccactgctac acagtttgtt catcatgctt tgaaagactc
     3121 tattttaaag ataatggaaa gcaagcagtc ttgtgagtta agtccatcaa agttagaaaa
     3181 aaatgaagat gtgaacacta atttaacaca cctattgaac atactttcag agcttgtgga
     3241 gaaaatattc atggcttcag aaatacttcc accgacattg agatatattt atgggtgttt
     3301 acagaaatct gttcagcata agtggcctac aaataccacc atgagaacaa gagttgttag
     3361 tggttttgtt tttcttcgac tcatctgtcc tgccatcctg aatccacgga tgttcaatat
     3421 catctcagat tctccatctc ctattgctgc aagaacactg atattagtgg ctaaatctgt
     3481 gcagaactta gcaaatcttg tggaatttgg agctaaggag ccctacatgg aaggtgtcaa
     3541 tccattcatc aaaagcaaca aacatcgtat gatcatgttt ttagatgaac ttgggaatgt
     3601 acctgaactt ccggacacta cagagcattc tagaacggac ctgtcccgtg atttagcagc
     3661 attgcatgag atttgcgtgg ctcattcaga tgaacttcga acgctcagta atgagcgtgg
     3721 tgcacagcag cacgtattga aaaagcttct ggctataaca gaactgcttc aacaaaaaca
     3781 aaaccagtat acaaaaacca atgatgtcag gtgattgcca actttcttgt acaaagttgg
     3841 cattataaga aagcattgct tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa
     3901 taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt acatggtcat
     3961 agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     4021 taaaaatata tcatcatgcc tcctctagac cagccaggac agaaatgcct cgacttcgct
     4081 gctacccaag gttgccgggt gacgcacacc gtggaaacgg atgaaggcac gaacccagtg
     4141 gacataagcc tgttcggttc gtaagctgta atgcaagtag cgtatgcgct cacgcaactg
     4201 gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt tcatggcttg
     4261 ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc aagcgcgtta
     4321 cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc agggcagtcg
     4381 ccctaaaaca aagttaaaca ttatgaggga agcggtgatc gccgaagtat cgactcaact
     4441 atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt
     4501 gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt tgctggttac
     4561 ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc ttttggaaac
     4621 ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca ttgttgtgca
     4681 cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg gagaatggca
     4741 gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg atctggctat
     4801 cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg cggaggaact
     4861 ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct taacgctatg
     4921 gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat
     4981 ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat
     5041 ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg cttatcttgg
     5101 acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg tccactacgt
     5161 gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg agccacccat gaccaaaatc
     5221 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5281 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5341 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5401 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5461 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5521 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5581 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5641 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5701 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5761 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5821 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5881 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
