LOCUS       sequence-66169-e        5741 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human KRas protein from chimeric cDNA sequence derived 
            primarily from HRAS codons with a KRAS 3' tail. Results in higher 
            than normal exogenous expression of KRas protein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5741)
  AUTHORS   Lampson BL, Pershing NL, Prinz JA, Lacsina JR, Marzluff WF, 
            Nicchitta CV, MacAlpine DM, Counter CM
  TITLE     Rare codons regulate KRas oncogenesis.
  JOURNAL   Curr Biol. 2013 Jan 7;23(1):70-5. doi: 10.1016/j.cub.2012.11.031. 
            Epub 2012 Dec 13.
  PUBMED    23246410
REFERENCE   2  (bases 1 to 5741)
  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"; doi: 
            "10.1016/j.cub.2012.11.031"; journalName: "Curr Biol"; date: 
            "2013-01-7- 7"; volume: "23"; issue: "1"; pages: "70-5"
COMMENT     Sequence Label: pBABE-Puro-KRas*
FEATURES             Location/Qualifiers
     source          1..5741
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             8..477
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    542..741
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             934..1353
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1321..1337
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1370..1393
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(1968..1988)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1973..2302
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2153..2288
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2312..2914
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWXXTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     primer_bind     complement(2312..2331)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2811..2831
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3013..3609
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3810..3827)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3981..4572)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4061..4080)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4750..5544,5545..5613))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVXXDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW"
     primer_bind     5373..5392
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5614..5718)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
       61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
      121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
      181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
      241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
      301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
      361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
      421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
      481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
      541 taagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
      601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
      661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
      721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
      781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
      841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt
      901 ctgactgtgt ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt
      961 ttgaccttag atcactggaa agatgtcgag nnncggctcg ctcacaacca gtcggtagat
     1021 gtcaagaaga gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga
     1081 tggccgcgag acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt
     1141 tcacctggcc cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg
     1201 gcttttgacc cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt
     1261 cttccatccg cgccgtctct cccccttgaa cctcctcttt cgaccccgcc tcaatcctcc
     1321 ctttatccag ccctcactcc ttctctaggc gccggccgga tccgccatgg actacaagga
     1381 cgacgatgac aagacggaat ataagcttgt ggtggtgggc gctggaggcg tgggaaagag
     1441 tgccctgacc atccagctga tccagaacca ctttgtggac gaatacgacc ccactataga
     1501 ggattcctac cggaagcagg tggtcattga tggggagacg tgcctgttgg acatcctgga
     1561 taccgccggc caggaggagt acagcgccat gcgggaccag tacatgcgca ccggggaggg
     1621 cttcctgtgt gtgtttgcca tcaacaacac caagtctttt gaggacatcc accattacag
     1681 ggagcagatc aaacgggtga aggactcgga ggacgtgccc atggtgctgg tggggaacaa
     1741 gtgtgacctg ccttcacgca ctgtggacac taagcaggct caggacctcg cccgaagcta
     1801 cggcatcccc ttcatcgaga cctcggccaa gacccggcag ggagtggatg atgccttcta
     1861 tacactagtt cgagaaattc gaaaacataa agaaaagatg agcaaagacg gtaaaaagaa
     1921 gaaaaagaag tcaaagacaa agtgtgtaat tatgtaagtc gacctgtgga atgtgtgtca
     1981 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     2041 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2101 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     2161 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     2221 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     2281 tggaggccta ggcttttgca aaaagcttac catgaccgag tacaagccca cggtgcgcct
     2341 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta
     2401 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca
     2461 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     2521 cgccgcggtg gcggtctgga nnnccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     2581 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     2641 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     2701 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     2761 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     2821 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gagtgcccga aggaccgcgc
     2881 gacctggtgc atgacccgca agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc
     2941 gaccgaaagg agcgcacgac cccatgcatc gataaaataa aagattttat ttagtctcca
     3001 gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc
     3061 attttgcaag gcatggaaaa atacataact gagaatagag aagttcagat caaggtcagg
     3121 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     3181 cggctcaggg ccaagaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt
     3241 aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc ggtccagccc
     3301 tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc
     3361 ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc
     3421 tccccgagct caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga
     3481 ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccnnngactt
     3541 gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg
     3601 tctttcattt ccgacttgtg gtctcgctgc cttgggaggg tctcctctga gtgattgact
     3661 acccgtcagc gggggtcttc acatgcagca tgtatcaaaa ttaatttggt tttttttctt
     3721 aagtatttac attaaatggc catagttgca ttaatgaatc ggccaacgcg cggggagagg
     3781 cggtttgcgt attggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3841 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3901 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3961 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4021 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4081 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4141 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     4201 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4261 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4321 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4381 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     4441 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4501 accaccgctg gtagcggtgg ttttnnnttt gtttgcaagc agcagattac gcgcagaaaa
     4561 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4621 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4681 ttaaattaaa aatgaagttt gcggccgcaa atcaatctaa agtatatatg agtaaacttg
     4741 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     4801 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     4861 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     4921 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     4981 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5041 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5101 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5161 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5221 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5281 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5341 accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt
     5401 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     5461 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttnnnt
     5521 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5581 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5641 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5701 acaaataggg gttccgcgca catttccccg aaaagtgcca c
//
