LOCUS       sequence-243323-        5683 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human Kras with a G12D 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  JOURNAL   Unpublished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            "Unpublished"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 G12D
FEATURES             Location/Qualifiers
     source          1..5683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             120..589
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(810..1409)
                     /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(893..913)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1390..1409
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(1419..1748)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1433..1568
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1733..1753
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(1765..2295,2296..2298,2299..2331))
                     /codon_start=1
                     /product="human oncoprotein generated by the G12V mutation 
                     in the small GTPase K-Ras"
                     /label=K-Ras (G12V)
                     /translation="MTEYKLVVVGADGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
                     DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
                     RKHKEKMSKDGKKKKKKSKTKCVIM"
     CDS             complement(1765..2331)
                     /codon_start=1
                     /product="human small GTPase proto-oncoprotein"
                     /label=K-Ras
                     /translation="MTEYKLVVVGADGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
                     DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
                     RKHKEKMSKDGKKKKKKSKTKCVIM"
     CDS             complement(2353..2379)
                     /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             complement(2359..2385)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(2408..2824)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(2424..2440)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(2456..2478)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    3025..3224
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             3289..3758
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        3789..3893
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3894..3962,3963..4754)
                     /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     complement(4112..4131)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4914..5502
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5403..5422
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5656..5673
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcaactatg
       61 gccatttaat gtaaatactt aagaaaaaaa accaaattaa ttttgataca tgctgcatgt
      121 gaaagacccc cgctgacggg tagtcaatca ctcagaggag accctcccaa ggaacagcga
      181 gaccacaagt cggatgcaac tgcaagaggg tttattggat acacgggtac ccgggcgact
      241 cagtcaatcg gaggactggc gccccgagtg aggggttgtg ggctctttta ttgagctcgg
      301 ggagcagaag cgcgcgaaca gaagcgagaa gcgaactgat tggttagttc aaataaggca
      361 cagggtcatt tcaggtcctt ggggcaccct ggaaacatct gatggttctc tagaaactgc
      421 tgagggctgg accgcatctg gggaccatct gttcttggcc ctgagccggg gcaggaactg
      481 cttaccacag atatcctgtt tggcccatat tcagctgttc catctgttct tggccctgag
      541 ccggggcagg aactgcttac cacagatatc ctgtttggcc catattcagc tgttccatct
      601 gttcctgacc ttgatctgaa cttctctatt ctcagttatg tatttttcca tgccttgcaa
      661 aatggcgtta cttaagctag cttgccaaac ctacaggtgg ggtctttcat tccccccttt
      721 ttctggagac taaataaaat cttttatttt atcgatgcat ggggtcgtgc gctcctttcg
      781 gtcgggcgct gcgggtcgtg gggcgggcgt caggcaccgg gcttgcgggt catgcaccag
      841 gtgcgcggtc cttcgggcac ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag
      901 aaggggaggt tgcggggcgc ggaggtctcc aggaaggcgg gcaccccggc gcgctcggcc
      961 gcctccactc cggggagcac gacggcgctg cccagaccct tgccctggtg gtcgggcgag
     1021 acgccgacgg tggccaggaa ccacgcgggc tccttgggcc ggtgcggcgc caggaggcct
     1081 tccatctgtt gctgcgcggc cagccgggaa ccgctcaact cggccatgcg cgggccgatc
     1141 tcggcgaaca ccgcccccgc ttcgacgctc tccggcgtgg tccagaccgc caccgcggcg
     1201 ccgtcgtccg cgacccacac cttgccgatg tcgagcccga cgcgcgtgag gaagagttct
     1261 tgcagctcgg tgacccgctc gatgtggcgg tccggatcga cggtgtggcg cgtggcgggg
     1321 tagtcggcga acgcggcggc gagggtgcgt acggccctgg ggacgtcgtc gcgggtggcg
     1381 aggcgcaccg tgggcttgta ctcggtcatg gtaagctttt tgcaaaagcc taggcctcca
     1441 aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc ggcctctgca
     1501 taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg
     1561 ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt
     1621 tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg
     1681 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac
     1741 tgacacacat tccacagggt cgacttacat aattacacac tttgtctttg acttcttttt
     1801 cttcttttta ccatctttgc tcatcttttc tttatgtttt cgaatttctc gaactaatgt
     1861 atagaaggca tcatcaacac cctgtcttgt ctttgctgat gtttcaataa aaggaattcc
     1921 ataacttctt gctaagtcct gagcctgttt tgtgtctact gttctggaag gcaaatcaca
     1981 tttatttcct actaggacca taggtacatc ttcagagtcc ttaactcttt taatttgttc
     2041 tctataatgg tgaatatctt caaatgattt agtattattt atggcaaata cacaaagaaa
     2101 gccctcccca gtcctcatgt actggtccct cattgcactg tactcctctt gacctgctgt
     2161 gtcgagaata tccaagagac aggtttctcc atcaattact acttgcttcc tgtaggaatc
     2221 ctctattgtt ggatcatatt cgtccacaaa atgattctga attagctgta tcgtcaaggc
     2281 actcttgcct acgccgtcag ctccaactac cacaagttta tattcagtca tggtggatcc
     2341 agagggtcct ccaagtgagg catagtcggg cacatcatat ggataacttg aagccattga
     2401 tccggccggc gcctagagaa ggagtgaggg ctggataaag ggaggatcga ggcggggtcg
     2461 aacgaggagg ttcaaggggg agagacgggg cggatggagg aagaggaggc ggaggcttag
     2521 ggtgtacaaa gggcttgacc cagggagggg ggtcaaaagc caaggcttcc caggtcacga
     2581 tgtaggggac ctggtctggg tgtccatgcg ggccaggtga aaagaccttg atcttaacct
     2641 gggtgatgag gtctcggtta aaggtgccgt ctcgcggcca tccgacgtta aaggttggcc
     2701 attctgcaga gcagaaggta acccaacgtc tcttcttgac atctaccgac tggttgtgag
     2761 cgatccgctc gacatctttc cagtgaccta aggtcaaact taagggagtg gtaacagtct
     2821 ggccctaatt ttcagacaaa tacagaaaca cagtcagaca gagacaacac agaacgatgc
     2881 tgcagcgctg cagcagacaa gacgcgcggc ttcggttcca aaccgaaagc aaaaattcag
     2941 acggaggcgg gaactgtttt aggttctcgt ctcctaccag aaccacatat cctgacgggg
     3001 tcggattcca catcgactcc cttcctcagg tcgggccaca aaaacggccc ccaaagtccc
     3061 tgggacgtct cccagggttg cggccgggtg ttcagaactc gtcagttcca ccacgggtcc
     3121 gccagataca gagctagtta gctaactagt accgacgcag gcgcataaaa tcagtcatag
     3181 acactagaca atcggacaga cacagataag ttgctggcca gcttgcctcc cggtggtggg
     3241 tcggtggtcc ctgggcaggg gtctcccgat cccggacgag cccccaaatg aaagaccccc
     3301 gctgacgggt agtcaatcac tcagaggaga ccctcccaag gaacagcgag accacaagtc
     3361 ggatgcaact gcaagagggt ttattggata cacgggtacc cgggcgactc agtcaatcgg
     3421 aggactggcg ccccgagtga ggggttgtgg gctcttttat tgagctcggg gagcagaagc
     3481 gcgcgaacag aagcgagaag cgaactgatt ggttagttca aataaggcac agggtcattt
     3541 caggtccttg gggcaccctg gaaacatctg atggttctct agaaactgct gagggctgga
     3601 ccgcatctgg ggaccatctg ttcttggccc tgagccgggg caggaactgc ttaccacaga
     3661 tatcctgttt ggcccatatt cagctgttcc atctgttctt ggccctgagc cggggcagga
     3721 actgcttacc acagatatcc tgtttggccc atattcaggc tgcaggtggc acttttcggg
     3781 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3841 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     3901 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3961 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4021 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4081 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4141 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4201 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4261 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4321 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4381 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4441 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4501 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4561 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4621 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4681 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4741 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttgcggc
     4801 cgcaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     4861 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4921 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4981 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5041 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5101 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5161 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5221 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5281 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5341 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5401 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5461 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5521 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     5581 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     5641 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aag
//
