LOCUS       sequence-170484-        5812 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of a constitutively active form of H-Ras.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5812)
  TITLE     Viral vectors.
REFERENCE   2  (bases 1 to 5812)
  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"
COMMENT     Sequence Label: pBABE puro H-Ras V12
FEATURES             Location/Qualifiers
     source          1..5812
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             17..486
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    551..750
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             951..1367
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1297..1319
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1335..1351
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             join(1422..1454,1455..1457,1458..1991)
                     /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"
     primer_bind     complement(2151..2171)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2156..2485
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2336..2471
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2495..3094
                     /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(2495..2514)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2991..3011
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3315..3784
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3914..3931)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4085..4673)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4165..4184)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4833..5624,5625..5693))
                     /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     5456..5475
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5694..5798)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aagtgccacc tgcagcctga atatgggcca aacaggatat ctgtggtaag cagttcctgc
       61 cccggctcag ggccaagaac agatggaaca gctgaatatg ggccaaacag gatatctgtg
      121 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtccagc
      181 cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga
      241 ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct
      301 gctccccgag ctcaataaaa gagcccacaa cccctcactc ggggcgccag tcctccgatt
      361 gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg
      421 tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt
      481 ctttcatttg ggggctcgtc cgggatcggg agacccctgc ccagggacca ccgacccacc
      541 accgggaggc aagctggcca gcaacttatc tgtgtctgtc cgattgtcta gtgtctatga
      601 ctgattttat gcgcctgcgt cggtactagt tagctaacta gctctgtatc tggcggaccc
      661 gtggtggaac tgacgagttc tgaacacccg gccgcaaccc tgggagacgt cccagggact
      721 ttgggggccg tttttgtggc ccgacctgag gaagggagtc gatgtggaat ccgaccccgt
      781 caggatatgt ggttctggta ggagacgaga acctaaaaca gttcccgcct ccgtctgaat
      841 ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct gctgcagcgc tgcagcatcg
      901 ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc tgaaaattag ggccagactg
      961 ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg atcgctcaca
     1021 accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca gaatggccaa
     1081 cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc acccaggtta
     1141 agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc tacatcgtga
     1201 cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta caccctaagc
     1261 ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct cgttcgaccc
     1321 cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccggc cggatcccag
     1381 tgtggtggta cagcttgggg ggccagggcc ggggccgagc gatgacggaa tataagctgg
     1441 tggtggtggg cgccgtcggt gtgggcaaga gtgcgctgac catccagctg atccagaacc
     1501 attttgtgga cgaatacgac cccactatag aggattccta ccggaagcag gtggtcattg
     1561 atggggagac gtgcctgttg gacatcctgg ataccgccgg ccaggaggag tacagcgcca
     1621 tgcgggacca gtacatgcgc accggggagg gcttcctgtg tgtgtttgcc atcaacaaca
     1681 ccaagtcttt tgaggacatc caccagtaca gggagcagat caaacgggtg aaggactcgg
     1741 atgacgtgcc catggtgctg gtggggaaca agtgtgacct ggctgcacgc actgtggaat
     1801 ctcggcaggc tcaggacctc gcccgaagct acggcatccc ctacatcgag acctcggcca
     1861 agacccggca gggagtggag gatgccttct acacgttggt gcgtgagatc cggcagcaca
     1921 agctgcggaa gctgaaccct cctgatgaga gtggccccgg ctgcatgagc tgcaagtgtg
     1981 tgctctcctg acgcagcaca agctcaggac atggaggtgc cggatgcagg aaggaggtgc
     2041 agacggaagg aggaggaagg aaggacggaa gcaaggaagg aaggaagggc tgctggagcc
     2101 cagtcacccc ccaagctgta ggaattcgcc agcacagtgg tcgaccctgt ggaatgtgtg
     2161 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2221 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2281 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     2341 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     2401 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     2461 ttttggaggc ctaggctttt gcaaaaagct taccatgacc gagtacaagc ccacggtgcg
     2521 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     2581 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     2641 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     2701 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     2761 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     2821 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     2881 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     2941 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     3001 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     3061 cacctggtgc atgacccgca agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc
     3121 gaccgaaagg agcgcacgac cccatgcatc gataaaataa aagattttat ttagtctcca
     3181 gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc
     3241 attttgcaag gcatggaaaa atacataact gagaatagag aagttcagat caaggtcagg
     3301 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     3361 cggctcaggg ccaagaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt
     3421 aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc ggtccagccc
     3481 tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc
     3541 ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc
     3601 tccccgagct caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga
     3661 ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg
     3721 gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct
     3781 ttcacatgca gcatgtatca aaattaattt ggtttttttt cttaagtatt tacattaaat
     3841 ggccatagtt gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3901 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3961 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4021 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4081 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4141 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4201 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4261 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4321 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4381 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4441 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4501 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4561 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4621 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4681 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4741 tggtcatgag attatcaaaa aggatcttca cctagatcct tttgcggccg caaatcaatc
     4801 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4861 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4921 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     4981 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5041 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5101 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     5161 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     5221 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     5281 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5341 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5401 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     5461 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5521 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5581 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     5641 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5701 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5761 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aa
//
