LOCUS       sequence-222764-        6367 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 6367)
  AUTHORS   Serrano M, Lin AW, McCurrach ME, Beach D, Lowe SW
  TITLE     Oncogenic ras provokes premature cell senescence associated with 
            accumulation of p53 and p16INK4a.
  JOURNAL   Cell. 1997 Mar 7. 88(5):593-602.
  PUBMED    9054499
REFERENCE   2  (bases 1 to 6367)
  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: "Cell. 1997 
            Mar 7. 88(5):593-602."
COMMENT     Sequence Label: pWZL hygro H-Ras V12
FEATURES             Location/Qualifiers
     source          1..6367
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             21..490
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    555..754
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             955..1371
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1301..1323
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1339..1355
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             join(1426..1458,1459..1461,1462..1995)
                     /codon_start=1
                     /product="human oncoprotein generated by the G12V mutation 
                     in the small GTPase H-Ras"
                     /label=H-Ras (G12V)
                     /translation="MTEYKLVVVGAVGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVV
                     IDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHQYREQIKRVK
                     DSDDVPMVLVGNKCDLAARTVESRQAQDLARSYGIPYIETSAKTRQGVEDAFYTLVREI
                     RQHKLRKLNPPDESGPGCMSCKCVLS"
     misc_feature    2194..2655
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2250..2267)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2477..2496
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2660..3685
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             2660..3685
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     LTR             3874..4343
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4473..4490)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4644..5232)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4724..4743)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5392..6183,6184..6252))
                     /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     6015..6034
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6253..6357)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgaaaagtgc cacctgcagc ctgaatatgg gccaaacagg atatctgtgg taagcagttc
       61 ctgccccggc tcagggccaa gaacagatgg aacagctgaa tatgggccaa acaggatatc
      121 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc
      181 cagccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa
      241 atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc
      301 ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggggcg ccagtcctcc
      361 gattgactga gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga
      421 cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg
      481 gggtctttca tttgggggct cgtccgggat cgggagaccc ctgcccaggg accaccgacc
      541 caccaccggg aggcaagctg gccagcaact tatctgtgtc tgtccgattg tctagtgtct
      601 atgactgatt ttatgcgcct gcgtcggtac tagttagcta actagctctg tatctggcgg
      661 acccgtggtg gaactgacga gttctgaaca cccggccgca accctgggag acgtcccagg
      721 gactttgggg gccgtttttg tggcccgacc tgaggaaggg agtcgatgtg gaatccgacc
      781 ccgtcaggat atgtggttct ggtaggagac gagaacctaa aacagttccc gcctccgtct
      841 gaatttttgc tttcggtttg gaaccgaagc cgcgcgtctt gtctgctgca gcgctgcagc
      901 atcgttctgt gttgtctctg tctgactgtg tttctgtatt tgtctgaaaa ttagggccag
      961 actgttacca ctcccttaag tttgacctta ggtcactgga aagatgtcga gcggatcgct
     1021 cacaaccagt cggtagatgt caagaagaga cgttgggtta ccttctgctc tgcagaatgg
     1081 ccaaccttta acgtcggatg gccgcgagac ggcaccttta accgagacct catcacccag
     1141 gttaagatca aggtcttttc acctggcccg catggacacc cagaccaggt cccctacatc
     1201 gtgacctggg aagccttggc ttttgacccc cctccctggg tcaagccctt tgtacaccct
     1261 aagcctccgc ctcctcttcc tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg
     1321 accccgcctc gatcctccct ttatccagcc ctcactcctt ctctaggcgc cggccggatc
     1381 ccagtgtggt ggtacagctt ggggggccag ggccggggcc gagcgatgac ggaatataag
     1441 ctggtggtgg tgggcgccgt cggtgtgggc aagagtgcgc tgaccatcca gctgatccag
     1501 aaccattttg tggacgaata cgaccccact atagaggatt cctaccggaa gcaggtggtc
     1561 attgatgggg agacgtgcct gttggacatc ctggataccg ccggccagga ggagtacagc
     1621 gccatgcggg accagtacat gcgcaccggg gagggcttcc tgtgtgtgtt tgccatcaac
     1681 aacaccaagt cttttgagga catccaccag tacagggagc agatcaaacg ggtgaaggac
     1741 tcggatgacg tgcccatggt gctggtgggg aacaagtgtg acctggctgc acgcactgtg
     1801 gaatctcggc aggctcagga cctcgcccga agctacggca tcccctacat cgagacctcg
     1861 gccaagaccc ggcagggagt ggaggatgcc ttctacacgt tggtgcgtga gatccggcag
     1921 cacaagctgc ggaagctgaa ccctcctgat gagagtggcc ccggctgcat gagctgcaag
     1981 tgtgtgctct cctgacgcag cacaagctca ggacatggag gtgccggatg caggaaggag
     2041 gtgcagacgg aaggaggagg aaggaaggac ggaagcaagg aaggaaggaa gggctgctgg
     2101 agcccagtca ccccccaagc tgtaggaatt cgccagcaca gtggtcgacc aattccggtt
     2161 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
     2221 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
     2281 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac
     2341 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
     2401 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
     2461 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca
     2521 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt
     2581 ttagtcgagg ttaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga
     2641 aaaacacgat gataatacca tgaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt
     2701 tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc
     2761 tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc
     2821 cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatcggccg cgctcccgat
     2881 tccggaagtg cttgacattg gggaatttag cgagagcctg acctattgca tctcccgccg
     2941 tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc
     3001 ggtcgcggag gccatggatg cgatcgctgc ggccgatctt agccagacga gcgggttcgg
     3061 cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat
     3121 tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt
     3181 cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct
     3241 cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt
     3301 cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt
     3361 ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc
     3421 ggagcttgca ggatcgccgc ggctccgggc gtatatgctc cgcattggtc ttgaccaact
     3481 ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga
     3541 cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc
     3601 ggccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag
     3661 cactcgtccg agggcaaagg aatagagtag atgccgaccg ggatctatcg ataaaataaa
     3721 agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca
     3781 agctagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaga
     3841 agttcagatc aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg
     3901 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc
     3961 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc
     4021 cccagatgcg gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc
     4081 ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct
     4141 tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggg
     4201 gcgccagtcc tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg
     4261 cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac
     4321 tacccgtcag cgggggtctt tcacatgcag catgtatcaa aattaatttg gttttttttc
     4381 ttaagtattt acattaaatg gccatagttg cattaatgaa tcggccaacg cgcggggaga
     4441 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4501 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4561 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4621 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4681 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4741 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4801 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4861 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4921 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4981 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5041 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5101 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5161 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5221 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5281 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5341 ttgcggccgc aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5401 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5461 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5521 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5581 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5641 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5701 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5761 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5821 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5881 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5941 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6001 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6061 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6121 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6181 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6241 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6301 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6361 atttccc
//
