LOCUS       sequence-158643-        5608 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human Kras with a S17N mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5608)
  TITLE     Mutant Ras Constructs
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5608)
  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            "Unpublished"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 S17N
FEATURES             Location/Qualifiers
     source          1..5608
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(63..80)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(234..822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(314..333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(982..1773,1774..1842))
                     /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     1605..1624
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1843..1947)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    2437..2636
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             2837..3253
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     3183..3205
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     3221..3237
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             3276..3302
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3282..3308
                     /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             3330..3896
                     /codon_start=1
                     /product="human small GTPase proto-oncoprotein"
                     /label=K-Ras
                     /translation="MTEYKLVVVGAGGVGKNALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
                     DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
                     RKHKEKMSKDGKKKKKKSKTKCVIM"
     primer_bind     complement(3908..3928)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3913..4242
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4093..4228
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4252..4851
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPAMRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4252..4271)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4748..4768
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5072..5541
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
       61 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
      121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
      181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
      241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
      301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
      361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
      421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
      481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
      541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
      601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
      661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
      721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
      781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      901 ttatcaaaaa ggatcttcac ctagatcctt ttgcggccgc aaatcaatct aaagtatata
      961 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     1021 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     1081 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     1141 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     1201 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     1261 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     1321 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     1381 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     1441 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     1501 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     1561 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     1621 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     1681 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     1741 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     1801 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     1861 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     1921 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgcagcctg
     1981 aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa
     2041 cagatggtcc ccagatgcgg tccagccctc agcagtttct agagaaccat cagatgtttc
     2101 cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg
     2161 cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc
     2221 tcactcgggg cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa
     2281 accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga
     2341 gtgattgact acccgtcagc gggggtcttt catttggggg ctcgtccggg atcgggagac
     2401 ccctgcccag ggaccaccga cccaccaccg ggaggcaagc tggccagcaa cttatctgtg
     2461 tctgtccgat tgtctagtgt ctatgactga ttttatgcgc ctgcgtcggt actagttagc
     2521 taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa cacccggccg
     2581 caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga cctgaggaag
     2641 ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag acgagaacct
     2701 aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa gccgcgcgtc
     2761 ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg tgtttctgta
     2821 tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct taggtcactg
     2881 gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga gacgttgggt
     2941 taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag acggcacctt
     3001 taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc cgcatggaca
     3061 cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc cccctccctg
     3121 ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg ccccgtctct
     3181 cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag ccctcactcc
     3241 ttctctaggc gccggccgga tcaatggctt caagttatcc atatgatgtg cccgactatg
     3301 cctcacttgg aggaccctct ggatccacca tgactgaata taaacttgtg gtagttggag
     3361 ctggcggcgt aggcaagaac gccttgacga tacagctaat tcagaatcat tttgtggacg
     3421 aatatgatcc aacaatagag gattcctaca ggaagcaagt agtaattgat ggagaaacct
     3481 gtctcttgga tattctcgac acagcaggtc aagaggagta cagtgcaatg agggaccagt
     3541 acatgaggac tggggagggc tttctttgtg tatttgccat aaataatact aaatcatttg
     3601 aagatattca ccattataga gaacaaatta aaagagttaa ggactctgaa gatgtaccta
     3661 tggtcctagt aggaaataaa tgtgatttgc cttccagaac agtagacaca aaacaggctc
     3721 aggacttagc aagaagttat ggaattcctt ttattgaaac atcagcaaag acaagacagg
     3781 gtgttgatga tgccttctat acattagttc gagaaattcg aaaacataaa gaaaagatga
     3841 gcaaagatgg taaaaagaag aaaaagaagt caaagacaaa gtgtgtaatt atgtaagtcg
     3901 accctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3961 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4021 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4081 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4141 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     4201 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagcttac catgaccgag
     4261 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc
     4321 gccgccgcgt tcgccgacta ccccgccatg cgccacaccg tcgatccgga ccgccacatc
     4381 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     4441 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     4501 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     4561 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     4621 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     4681 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     4741 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     4801 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg acgcccgccc
     4861 cacgacccgc agcgcccgac cgaaaggagc gcacgacccc atgcatcgat aaaataaaag
     4921 attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag
     4981 ctagcttaag taacgccatt ttgcaaggca tggaaaaata cataactgag aatagagaag
     5041 ttcagatcaa ggtcaggaac agatggaaca gctgaatatg ggccaaacag gatatctgtg
     5101 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gaacagctga atatgggcca
     5161 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc
     5221 cagatgcggt ccagccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc
     5281 aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc
     5341 tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggggc
     5401 gccagtcctc cgattgactg agtcgcccgg gtacccgtgt atccaataaa ccctcttgca
     5461 gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta
     5521 cccgtcagcg ggggtctttc acatgcagca tgtatcaaaa ttaatttggt tttttttctt
     5581 aagtatttac attaaatggc catagttg
//
