LOCUS       sequence-198776-        5668 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5668)
  AUTHORS   Khosravi-Far R, White MA, Westwick JK, Solski PA, 
            Chrzanowska-Wodnicka M, Van Aelst L, Wigler MH, Der CJ
  TITLE     Oncogenic Ras activation of Raf/mitogen-activated protein 
            kinase-independent pathways is sufficient to cause tumorigenic 
            transformation.
  JOURNAL   Mol Cell Biol. 1996 Jul . 16(7):3923-33.
  PUBMED    8668210
REFERENCE   2  (bases 1 to 5668)
  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: "Mol Cell 
            Biol. 1996 Jul . 16(7):3923-33."
COMMENT     Sequence Label: pBabe K-Ras 12V
FEATURES             Location/Qualifiers
     source          1..5668
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        338..442
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(443..511,512..1303)
                     /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(661..680)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1463..2051
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1952..1971
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2205..2222
                     /label=L4440
                     /note="L4440 vector, forward primer"
     LTR             2352..2821
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(3042..3641)
                     /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(3125..3145)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     3622..3641
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(3651..3980)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3665..3800
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3965..3985
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(4045..4575,4576..4578,4579..4611))
                     /codon_start=1
                     /product="human oncoprotein generated by the G12V mutation 
                     in the small GTPase K-Ras"
                     /label=K-Ras (G12V)
                     /translation="MTEYKLVVVGAVGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVK
                     DSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQGVDDAFYTLVREI
                     RKHKEKMSKDGKKKKKKSKTKCVIM"
     CDS             complement(4625..5041)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(4641..4657)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(4673..4695)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    5242..5441
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             join(5506..5668,1..307)
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 ctcttttatt gagctcgggg agcagaagcg cgcgaacaga agcgagaagc gaactgattg
       61 gttagttcaa ataaggcaca gggtcatttc aggtccttgg ggcaccctgg aaacatctga
      121 tggttctcta gaaactgctg agggctggac cgcatctggg gaccatctgt tcttggccct
      181 gagccggggc aggaactgct taccacagat atcctgtttg gcccatattc agctgttcca
      241 tctgttcttg gccctgagcc ggggcaggaa ctgcttacca cagatatcct gtttggccca
      301 tattcaggct gcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
      361 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
      421 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
      481 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
      541 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
      601 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
      661 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
      721 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
      781 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
      841 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
      901 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
      961 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     1021 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     1081 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     1141 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     1201 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     1261 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     1321 ctcatatata ctttagattg atttgcggcc gcaaaaggat ctaggtgaag atcctttttg
     1381 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     1441 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     1501 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     1561 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     1621 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     1681 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     1741 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     1801 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     1861 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     1921 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     1981 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     2041 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     2101 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     2161 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     2221 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     2281 aatgcaacta tggccattta atgtaaatac ttaagaaaaa aaaccaaatt aattttgata
     2341 catgctgcat gtgaaagacc cccgctgacg ggtagtcaat cactcagagg agaccctccc
     2401 aaggaacagc gagaccacaa gtcggatgca actgcaagag ggtttattgg atacacgggt
     2461 acccgggcga ctcagtcaat cggaggactg gcgccccgag tgaggggttg tgggctcttt
     2521 tattgagctc ggggagcaga agcgcgcgaa cagaagcgag aagcgaactg attggttagt
     2581 tcaaataagg cacagggtca tttcaggtcc ttggggcacc ctggaaacat ctgatggttc
     2641 tctagaaact gctgagggct ggaccgcatc tggggaccat ctgttcttgg ccctgagccg
     2701 gggcaggaac tgcttaccac agatatcctg tttggcccat attcagctgt tccatctgtt
     2761 cttggccctg agccggggca ggaactgctt accacagata tcctgtttgg cccatattca
     2821 gctgttccat ctgttcctga ccttgatctg aacttctcta ttctcagtta tgtatttttc
     2881 catgccttgc aaaatggcgt tacttaagct agcttgccaa acctacaggt ggggtctttc
     2941 attcccccct ttttctggag actaaataaa atcttttatt ttatcgatgc atggggtcgt
     3001 gcgctccttt cggtcgggcg ctgcgggtcg tggggcgggc gtcaggcacc gggcttgcgg
     3061 gtcatgcacc aggtgcgcgg tccttcgggc acctcgacgt cggcggtgac ggtgaagccg
     3121 agccgctcgt agaaggggag gttgcggggc gcggaggtct ccaggaaggc gggcaccccg
     3181 gcgcgctcgg ccgcctccac tccggggagc acgacggcgc tgcccagacc cttgccctgg
     3241 tggtcgggcg agacgccgac ggtggccagg aaccacgcgg gctccttggg ccggtgcggc
     3301 gccaggaggc cttccatctg ttgctgcgcg gccagccggg aaccgctcaa ctcggccatg
     3361 cgcgggccga tctcggcgaa caccgccccc gcttcgacgc tctccggcgt ggtccagacc
     3421 gccaccgcgg cgccgtcgtc cgcgacccac accttgccga tgtcgagccc gacgcgcgtg
     3481 aggaagagtt cttgcagctc ggtgacccgc tcgatgtggc ggtccggatc gacggtgtgg
     3541 cgcgtggcgg ggtagtcggc gaacgcggcg gcgagggtgc gtacggccct ggggacgtcg
     3601 tcgcgggtgg cgaggcgcac cgtgggcttg tactcggtca tggtaagctt tttgcaaaag
     3661 cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc cgaggcggcc
     3721 tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg ggcggaactg
     3781 ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg
     3841 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acacctggtt
     3901 gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt
     3961 ccacacccta actgacacac attccacagg gtcgaccact gtgctggcga attcctacgt
     4021 accaccacac tgggatccgt cgacttacat aattacacac tttgtctttg acttcttttt
     4081 cttcttttta ccatctttgc tcatcttttc tttatgtttt cgaatttctc gaactaatgt
     4141 atagaaggca tcatcaacac cctgtcttgt ctttgctgat gtttcaataa aaggaattcc
     4201 ataacttctt gctaagtcct gagcctgttt tgtgtctact gttctggaag gcaaatcaca
     4261 tttatttcct actaggacca taggtacatc ttcagagtcc ttaactcttt taatttgttc
     4321 tctataatgg tgaatatctt caaatgattt agtattattt atggcaaata cacaaagaaa
     4381 gccctcccca gtcctcatgt actggtccct cattgcactg tactcctctt gacctgctgt
     4441 gtcgagaata tccaagagac aggtttctcc atcaattact acttgcttcc tgtaggaatc
     4501 ctctattgtt ggatcatatt cgtccacaaa atgattctga attagctgta tcgtcaaggc
     4561 actcttgcct acgccaacag ctccaactac cacaagttta tattcagtca tggtggatcc
     4621 ggccggcgcc tagagaagga gtgagggctg gataaaggga ggatcgaggc ggggtcgaac
     4681 gaggaggttc aagggggaga gacggggcgg atggaggaag aggaggcgga ggcttagggt
     4741 gtacaaaggg cttgacccag ggaggggggt caaaagccaa ggcttcccag gtcacgatgt
     4801 aggggacctg gtctgggtgt ccatgcgggc caggtgaaaa gaccttgatc ttaacctggg
     4861 tgatgaggtc tcggttaaag gtgccgtctc gcggccatcc gacgttaaag gttggccatt
     4921 ctgcagagca gaaggtaacc caacgtctct tcttgacatc taccgactgg ttgtgagcga
     4981 tccgctcgac atctttccag tgacctaagg tcaaacttaa gggagtggta acagtctggc
     5041 cctaattttc agacaaatac agaaacacag tcagacagag acaacacaga acgatgctgc
     5101 agcgctgcag cagacaagac gcgcggcttc ggttccaaac cgaaagcaaa aattcagacg
     5161 gaggcgggaa ctgttttagg ttctcgtctc ctaccagaac cacatatcct gacggggtcg
     5221 gattccacat cgactccctt cctcaggtcg ggccacaaaa acggccccca aagtccctgg
     5281 gacgtctccc agggttgcgg ccgggtgttc agaactcgtc agttccacca cgggtccgcc
     5341 agatacagag ctagttagct aactagtacc gacgcaggcg cataaaatca gtcatagaca
     5401 ctagacaatc ggacagacac agataagttg ctggccagct tgcctcccgg tggtgggtcg
     5461 gtggtccctg ggcaggggtc tcccgatccc ggacgagccc ccaaatgaaa gacccccgct
     5521 gacgggtagt caatcactca gaggagaccc tcccaaggaa cagcgagacc acaagtcgga
     5581 tgcaactgca agagggttta ttggatacac gggtacccgg gcgactcagt caatcggagg
     5641 actggcgccc cgagtgaggg gttgtggg
//
