LOCUS       sequence-158137-        3374 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), G12V mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3374)
  AUTHORS   Esposito
  TITLE     RAS mutant collection V2.0
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 3374)
  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            "unpublished"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 G12V
FEATURES             Location/Qualifiers
     source          1..3374
                     /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(754..853)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(870..889)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(871..889)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(894..910)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(894..910)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1341..2132
                     /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      2225..2813
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2714..2733
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2967..2984
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3143..3170)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3262..3348)
                     /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 aattgatgag caatgctttt ttataatgcc aactttgtac aaaaaagttg
      181 gcaccatgac tgagtacaaa ctggtggtgg ttggagcagt tggtgttggg aaaagcgcac
      241 tgacaatcca gctaatccag aaccactttg tagatgaata tgatcccacc atagaggatt
      301 cttacagaaa acaagtggtt atagatggtg aaacctgttt gttggacata ctggatacag
      361 ctggacaaga agagtacagt gccatgagag accaatacat gaggacaggc gaaggcttcc
      421 tctgtgtatt tgccatcaat aatagcaagt catttgcgga tattaacctc tacagggagc
      481 agattaagcg agtaaaagac tcggatgatg tacctatggt gctagtggga aacaagtgtg
      541 atttgccaac aaggacagtt gatacaaaac aagcccacga actggccaag agttacggga
      601 ttccattcat tgaaacctca gccaagacca gacagggtgt tgaagatgct ttttacacac
      661 tggtaagaga aatacgccag taccgaatga aaaaactcaa cagcagtgat gatgggactc
      721 agggttgtat gggattgcca tgtgtggtga tgtaatcaac tttcttgtac aaagttggca
      781 ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata
      841 aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac atggtcatag
      901 ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata
      961 aaaatatatc atcatgcctc ctctagacca gccaggacag aaatgcctcg acttcgctgc
     1021 tacccaaggt tgccgggtga cgcacaccgt ggaaacggat gaaggcacga acccagtgga
     1081 cataagcctg ttcggttcgt aagctgtaat gcaagtagcg tatgcgctca cgcaactggt
     1141 ccagaacctt gaccgaacgc agcggtggta acggcgcagt ggcggttttc atggcttgtt
     1201 atgactgttt ttttggggta cagtctatgc ctcgggcatc caagcagcaa gcgcgttacg
     1261 ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt tacgcagcag ggcagtcgcc
     1321 ctaaaacaaa gttaaacatt atgagggaag cggtgatcgc cgaagtatcg actcaactat
     1381 cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc gtacatttgt
     1441 acggctccgc agtggatggc ggcctgaagc cacacagtga tattgatttg ctggttacgg
     1501 tgaccgtaag gcttgatgaa acaacgcggc gagctttgat caacgacctt ttggaaactt
     1561 cggcttcccc tggagagagc gagattctcc gcgctgtaga agtcaccatt gttgtgcacg
     1621 acgacatcat tccgtggcgt tatccagcta agcgcgaact gcaatttgga gaatggcagc
     1681 gcaatgacat tcttgcaggt atcttcgagc cagccacgat cgacattgat ctggctatct
     1741 tgctgacaaa agcaagagaa catagcgttg ccttggtagg tccagcggcg gaggaactct
     1801 ttgatccggt tcctgaacag gatctatttg aggcgctaaa tgaaacctta acgctatgga
     1861 actcgccgcc cgactgggct ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt
     1921 ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg
     1981 agcgcctgcc ggcccagtat cagcccgtca tacttgaagc tagacaggct tatcttggac
     2041 aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc cactacgtga
     2101 aaggcgagat caccaaggta gtcggcaaat aaccctcgag ccacccatga ccaaaatccc
     2161 ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     2221 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     2281 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     2341 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     2401 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     2461 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2521 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2581 cgacctacac cgaactgaga tacctacagc gtgagcattg agaaagcgcc acgcttcccg
     2641 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2701 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2761 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2821 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     2881 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     2941 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     3001 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     3061 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt accgctagcc
     3121 aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttagtttg
     3181 atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac
     3241 gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
     3301 gataaaacga aaggcccagt cttccgactg agcctttcgt tttatttgat gcctggcagt
     3361 tccctactct cgcg
//
