LOCUS       sequence-242434-        5683 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human Kras with a G12C mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5683)
  TITLE     Mutant Ras Constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5683)
  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            "Unpublished"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: pBabe-Kras G12C
FEATURES             Location/Qualifiers
     source          1..5683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             14..483
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    548..747
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             948..1364
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1294..1316
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1332..1348
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1387..1413
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1393..1419
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             1441..2007
                     /codon_start=1
                     /product="human small GTPase proto-oncoprotein"
                     /label=K-Ras
                     /translation="MTEYKLVVVGACGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
                     DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
                     RKHKEKMSKDGKKKKKKSKTKCVIM"
     primer_bind     complement(2019..2039)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2024..2353
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2204..2339
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2363..2962
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2363..2382)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2859..2879
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3183..3652
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3782..3799)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3953..4541)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4033..4052)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4701..5492,5493..5561))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5324..5343
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5562..5666)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgccacctgc agcctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc
       61 ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat atctgtggta
      121 agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg gtccagccct
      181 cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc
      241 tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct
      301 ccccgagctc aataaaagag cccacaaccc ctcactcggg gcgccagtcc tccgattgac
      361 tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc cgacttgtgg
      421 tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag cgggggtctt
      481 tcatttgggg gctcgtccgg gatcgggaga cccctgccca gggaccaccg acccaccacc
      541 gggaggcaag ctggccagca acttatctgt gtctgtccga ttgtctagtg tctatgactg
      601 attttatgcg cctgcgtcgg tactagttag ctaactagct ctgtatctgg cggacccgtg
      661 gtggaactga cgagttctga acacccggcc gcaaccctgg gagacgtccc agggactttg
      721 ggggccgttt ttgtggcccg acctgaggaa gggagtcgat gtggaatccg accccgtcag
      781 gatatgtggt tctggtagga gacgagaacc taaaacagtt cccgcctccg tctgaatttt
      841 tgctttcggt ttggaaccga agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc
      901 tgtgttgtct ctgtctgact gtgtttctgt atttgtctga aaattagggc cagactgtta
      961 ccactccctt aagtttgacc ttaggtcact ggaaagatgt cgagcggatc gctcacaacc
     1021 agtcggtaga tgtcaagaag agacgttggg ttaccttctg ctctgcagaa tggccaacct
     1081 ttaacgtcgg atggccgcga gacggcacct ttaaccgaga cctcatcacc caggttaaga
     1141 tcaaggtctt ttcacctggc ccgcatggac acccagacca ggtcccctac atcgtgacct
     1201 gggaagcctt ggcttttgac ccccctccct gggtcaagcc ctttgtacac cctaagcctc
     1261 cgcctcctct tcctccatcc gccccgtctc tcccccttga acctcctcgt tcgaccccgc
     1321 ctcgatcctc cctttatcca gccctcactc cttctctagg cgccggccgg atcaatggct
     1381 tcaagttatc catatgatgt gcccgactat gcctcacttg gaggaccctc tggatccacc
     1441 atgactgaat ataaacttgt ggtagttgga gcttgcggcg taggcaagag tgccttgacg
     1501 atacagctaa ttcagaatca ttttgtggac gaatatgatc caacaataga ggattcctac
     1561 aggaagcaag tagtaattga tggagaaacc tgtctcttgg atattctcga cacagcaggt
     1621 caagaggagt acagtgcaat gagggaccag tacatgagga ctggggaggg ctttctttgt
     1681 gtatttgcca taaataatac taaatcattt gaagatattc accattatag agaacaaatt
     1741 aaaagagtta aggactctga agatgtacct atggtcctag taggaaataa atgtgatttg
     1801 ccttccagaa cagtagacac aaaacaggct caggacttag caagaagtta tggaattcct
     1861 tttattgaaa catcagcaaa gacaagacag ggtgttgatg atgccttcta tacattagtt
     1921 cgagaaattc gaaaacataa agaaaagatg agcaaagatg gtaaaaagaa gaaaaagaag
     1981 tcaaagacaa agtgtgtaat tatgtaagtc gaccctgtgg aatgtgtgtc agttagggtg
     2041 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     2101 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2161 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     2221 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     2281 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     2341 aggcttttgc aaaaagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg
     2401 cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac
     2461 gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt
     2521 cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt
     2581 ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc
     2641 gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct
     2701 ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga
     2761 ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg
     2821 cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg
     2881 gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat
     2941 gacccgcaag cccggtgcct gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag
     3001 cgcacgaccc catgcatcga taaaataaaa gattttattt agtctccaga aaaagggggg
     3061 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
     3121 atggaaaaat acataactga gaatagagaa gttcagatca aggtcaggaa cagatggaac
     3181 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     3241 aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg
     3301 ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct
     3361 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
     3421 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
     3481 ataaaagagc ccacaacccc tcactcgggg cgccagtcct ccgattgact gagtcgcccg
     3541 ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc
     3601 cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt cacatgcagc
     3661 atgtatcaaa attaatttgg ttttttttct taagtattta cattaaatgg ccatagttgc
     3721 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     3781 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3841 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3901 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3961 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4021 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4081 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4141 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4201 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4261 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4321 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4381 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4441 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4501 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4561 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4621 tatcaaaaag gatcttcacc tagatccttt tgcggccgca aatcaatcta aagtatatat
     4681 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4741 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4801 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4861 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4921 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4981 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5041 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5101 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5161 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5221 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5281 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5341 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5401 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5461 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5521 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5581 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5641 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aag
//
