LOCUS       sequence-158146-        3263 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway ORF Entry clone of human HRAS [NM_005343.2 ] 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 3263)
  AUTHORS   Esposito
  TITLE     RAS mutant collection V2.0
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 3263)
  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            "unpublished"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.HRAS G12D
FEATURES             Location/Qualifiers
     source          1..3263
                     /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"
     CDS             join(186..218,219..221,222..755)
                     /codon_start=1
                     /product="human oncoprotein generated by the G12V mutation 
                     in the small GTPase H-Ras"
                     /label=H-Ras (G12V)
                     /translation="MTEYKLVVVGADGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHQYREQIKRVK
                     DSDDVPMVLVGNKCDLAARTVESRQAQDLARSYGIPYIETSAKTRQGVEDAFYTLVREI
                     RQHKLRKLNPPDESGPGCMSCKCVLS"
     CDS             186..755
                     /codon_start=1
                     /product="human small GTPase proto-oncoprotein"
                     /label=H-Ras
                     /translation="MTEYKLVVVGADGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHQYREQIKRVK
                     DSDDVPMVLVGNKCDLAARTVESRQAQDLARSYGIPYIETSAKTRQGVEDAFYTLVREI
                     RQHKLRKLNPPDESGPGCMSCKCVLS"
     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"
     CDS             1230..2021
                     /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      2114..2702
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2603..2622
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2856..2873
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3032..3059)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3151..3237)
                     /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 gcaccatgac ggaatataag ctggtggtgg tgggcgccga cggtgtgggc aagagtgcgc
      241 tgaccatcca gctgatccag aaccattttg tggacgaata cgaccccact atagaggatt
      301 cctaccggaa gcaggtggtc attgatgggg agacgtgcct gttggacatc ctggataccg
      361 ccggccagga ggagtacagc gccatgcggg accagtacat gcgcaccggg gagggcttcc
      421 tgtgtgtgtt tgccatcaac aacaccaagt cttttgagga catccaccag tacagggagc
      481 agatcaaacg ggtgaaggac tcggatgacg tgcccatggt gctggtgggg aacaagtgtg
      541 acctggctgc acgcactgtg gaatctcggc aggctcagga cctcgcccga agctacggca
      601 tcccctacat cgagacctcg gccaagaccc ggcagggagt ggaggatgcc ttctacacgt
      661 tggtgcgtga gatccggcag cacaagctgc ggaagctgaa ccctcctgat gagagtggcc
      721 ccggctgcat gagctgcaag tgtgtgctct cctaatcaac tttcttgtac aaagttggca
      781 ttataggaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata
      841 aaatcattat ttgcatgcaa gcttagatcc tctagaccag ccaggacaga aatgcctcga
      901 cttcgctgct acccaaggtt gccgggtgac gcacaccgtg gaaacggatg aaggcacgaa
      961 cccagtggac ataagcctgt tcggttcgta agctgtaatg caagtagcgt atgcgctcac
     1021 gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca
     1081 tggcttgtta tgactgtttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag
     1141 cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg
     1201 gcagtcgccc taaaacaaag ttaaacatta tgagggaagc ggtgatcgcc gaagtatcga
     1261 ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg
     1321 tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc
     1381 tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt
     1441 tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg
     1501 ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag
     1561 aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc
     1621 tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg
     1681 aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa
     1741 cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt
     1801 cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact
     1861 gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt
     1921 atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc
     1981 actacgtgaa aggcgagatc accaaggtag tcggcaaata accctcgagc cacccatgac
     2041 caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga
     2101 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2161 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     2221 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     2281 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     2341 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     2401 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     2461 tggagcgaac gacctacacc gaactgagat acctacagcg tgagcattga gaaagcgcca
     2521 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     2581 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     2641 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     2701 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     2761 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     2821 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     2881 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     2941 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta
     3001 ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg
     3061 cttagtttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg
     3121 cttcacaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag cgttcaccga
     3181 caaacaacag ataaaacgaa aggcccagtc ttccgactga gcctttcgtt ttatttgatg
     3241 cctggcagtt ccctactctc gcg
//
