LOCUS       sequence-158143-        3287 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), Q61K mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3287)
  AUTHORS   Esposito
  TITLE     RAS mutant collection V2.0
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 3287)
  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            "unpublished"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 Q61K
FEATURES             Location/Qualifiers
     source          1..3287
                     /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(778..877)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1254..2045
                     /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      2138..2726
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2627..2646
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2880..2897
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3056..3083)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3175..3261)
                     /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 gcagtgataa gggaggctct gccgccacca tgactgagta caaactggtg gtggttggag
      241 caggtggtgt tgggaaaagc gcactgacaa tccagctaat ccagaaccac tttgtagatg
      301 aatatgatcc caccatagag gattcttaca gaaaacaagt ggttatagat ggtgaaacct
      361 gtttgttgga catactggat acagctggaa aagaagagta cagtgccatg agagaccaat
      421 acatgaggac aggcgaaggc ttcctctgtg tatttgccat caataatagc aagtcatttg
      481 cggatattaa cctctacagg gagcagatta agcgagtaaa agactcggat gatgtaccta
      541 tggtgctagt gggaaacaag tgtgatttgc caacaaggac agttgataca aaacaagccc
      601 acgaactggc caagagttac gggattccat tcattgaaac ctcagccaag accagacagg
      661 gtgttgaaga tgctttttac acactggtaa gagaaatacg ccagtaccga atgaaaaaac
      721 tcaacagcag tgatgatggg actcagggtt gtatgggatt gccatgtgtg gtgatgtaat
      781 caactttctt gtacaaagtt ggcattatag gaaagcattg cttatcaatt tgttgcaacg
      841 aacaggtcac tatcagtcaa aataaaatca ttatttgcat gcaagcttag atcctctaga
      901 ccagccagga cagaaatgcc tcgacttcgc tgctacccaa ggttgccggg tgacgcacac
      961 cgtggaaacg gatgaaggca cgaacccagt ggacataagc ctgttcggtt cgtaagctgt
     1021 aatgcaagta gcgtatgcgc tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg
     1081 gtaacggcgc agtggcggtt ttcatggctt gttatgactg tttttttggg gtacagtcta
     1141 tgcctcgggc atccaagcag caagcgcgtt acgccgtggg tcgatgtttg atgttatgga
     1201 gcagcaacga tgttacgcag cagggcagtc gccctaaaac aaagttaaac attatgaggg
     1261 aagcggtgat cgccgaagta tcgactcaac tatcagaggt agttggcgtc atcgagcgcc
     1321 atctcgaacc gacgttgctg gccgtacatt tgtacggctc cgcagtggat ggcggcctga
     1381 agccacacag tgatattgat ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc
     1441 ggcgagcttt gatcaacgac cttttggaaa cttcggcttc ccctggagag agcgagattc
     1501 tccgcgctgt agaagtcacc attgttgtgc acgacgacat cattccgtgg cgttatccag
     1561 ctaagcgcga actgcaattt ggagaatggc agcgcaatga cattcttgca ggtatcttcg
     1621 agccagccac gatcgacatt gatctggcta tcttgctgac aaaagcaaga gaacatagcg
     1681 ttgccttggt aggtccagcg gcggaggaac tctttgatcc ggttcctgaa caggatctat
     1741 ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc gcccgactgg gctggcgatg
     1801 agcgaaatgt agtgcttacg ttgtcccgca tttggtacag cgcagtaacc ggcaaaatcg
     1861 cgccgaagga tgtcgctgcc gactgggcaa tggagcgcct gccggcccag tatcagcccg
     1921 tcatacttga agctagacag gcttatcttg gacaagaaga agatcgcttg gcctcgcgcg
     1981 cagatcagtt ggaagaattt gtccactacg tgaaaggcga gatcaccaag gtagtcggca
     2041 aataaccctc gagccaccca tgaccaaaat cccttaacgt gagttacgcg tcgttccact
     2101 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     2161 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     2221 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     2281 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     2341 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     2401 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     2461 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2521 agcgtgagca ttgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     2581 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     2641 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     2701 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     2761 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     2821 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     2881 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     2941 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     3001 agcgcaacgc aattaatacg cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag
     3061 gccatccgtc aggatggcct tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc
     3121 tgcccgccac cctccgggcc gttgcttcac aacgttcaaa tccgctcccg gcggatttgt
     3181 cctactcagg agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtcttccga
     3241 ctgagccttt cgttttattt gatgcctggc agttccctac tctcgcg
//
