LOCUS       sequence-158139-        3284 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway ORF Entry clone of human NRAS [NM_002524.4 ] with stop codon
            (for native or N-terminal fusions), G12D mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3284)
  AUTHORS   Esposito
  TITLE     RAS mutant collection V2.0
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 3284)
  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"; journalName: 
            "unpublished"
COMMENT     Sequence Label: Hs.NRAS G12D
FEATURES             Location/Qualifiers
     source          1..3284
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..45
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     37..54
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     38..54
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    84..183
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(775..874)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1251..2042
                     /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      2135..2723
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2624..2643
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2877..2894
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3053..3080)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3172..3258)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc cagtcttaaa
       61 attcgagctc ggtccgagga tccgaaataa tgattttatt ttgactgata gtgacctgtt
      121 cgttgcaaca aattgataag caatgctttt ttataatgcc aactttgtac aaaaaagttg
      181 gcaggagaga tggctctgcc gccaccatga ctgagtacaa actggtggtg gttggagcag
      241 atggtgttgg gaaaagcgca ctgacaatcc agctaatcca gaaccacttt gtagatgaat
      301 atgatcccac catagaggat tcttacagaa aacaagtggt tatagatggt gaaacctgtt
      361 tgttggacat actggataca gctggacaag aagagtacag tgccatgaga gaccaataca
      421 tgaggacagg cgaaggcttc ctctgtgtat ttgccatcaa taatagcaag tcatttgcgg
      481 atattaacct ctacagggag cagattaagc gagtaaaaga ctcggatgat gtacctatgg
      541 tgctagtggg aaacaagtgt gatttgccaa caaggacagt tgatacaaaa caagcccacg
      601 aactggccaa gagttacggg attccattca ttgaaacctc agccaagacc agacagggtg
      661 ttgaagatgc tttttacaca ctggtaagag aaatacgcca gtaccgaatg aaaaaactca
      721 acagcagtga tgatgggact cagggttgta tgggattgcc atgtgtggtg atgtaatcaa
      781 ctttcttgta caaagttggc attataggaa agcattgctt atcaatttgt tgcaacgaac
      841 aggtcactat cagtcaaaat aaaatcatta tttgcatgca agcttagatc ctctagacca
      901 gccaggacag aaatgcctcg acttcgctgc tacccaaggt tgccgggtga cgcacaccgt
      961 ggaaacggat gaaggcacga acccagtgga cataagcctg ttcggttcgt aagctgtaat
     1021 gcaagtagcg tatgcgctca cgcaactggt ccagaacctt gaccgaacgc agcggtggta
     1081 acggcgcagt ggcggttttc atggcttgtt atgactgttt ttttggggta cagtctatgc
     1141 ctcgggcatc caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca
     1201 gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa gttaaacatt atgagggaag
     1261 cggtgatcgc cgaagtatcg actcaactat cagaggtagt tggcgtcatc gagcgccatc
     1321 tcgaaccgac gttgctggcc gtacatttgt acggctccgc agtggatggc ggcctgaagc
     1381 cacacagtga tattgatttg ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc
     1441 gagctttgat caacgacctt ttggaaactt cggcttcccc tggagagagc gagattctcc
     1501 gcgctgtaga agtcaccatt gttgtgcacg acgacatcat tccgtggcgt tatccagcta
     1561 agcgcgaact gcaatttgga gaatggcagc gcaatgacat tcttgcaggt atcttcgagc
     1621 cagccacgat cgacattgat ctggctatct tgctgacaaa agcaagagaa catagcgttg
     1681 ccttggtagg tccagcggcg gaggaactct ttgatccggt tcctgaacag gatctatttg
     1741 aggcgctaaa tgaaacctta acgctatgga actcgccgcc cgactgggct ggcgatgagc
     1801 gaaatgtagt gcttacgttg tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc
     1861 cgaaggatgt cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca
     1921 tacttgaagc tagacaggct tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag
     1981 atcagttgga agaatttgtc cactacgtga aaggcgagat caccaaggta gtcggcaaat
     2041 aaccctcgag ccacccatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag
     2101 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     2161 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     2221 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     2281 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     2341 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     2401 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     2461 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     2521 gtgagcattg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     2581 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     2641 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     2701 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     2761 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     2821 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     2881 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     2941 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     3001 gcaacgcaat taatacgcgt accgctagcc aggaagagtt tgtagaaacg caaaaaggcc
     3061 atccgtcagg atggccttct gcttagtttg atgcctggca gtttatggcg ggcgtcctgc
     3121 ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg gatttgtcct
     3181 actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt cttccgactg
     3241 agcctttcgt tttatttgat gcctggcagt tccctactct cgcg
//
