LOCUS       sequence-162793-        5812 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 5812)
  TITLE     pBABE-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 Ras V12
FEATURES             Location/Qualifiers
     source          1..5812
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             68..537
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(758..1357)
                     /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(841..861)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1338..1357
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(1367..1696)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1381..1516
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1681..1701
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(1861..2394,2395..2397,2398..2430))
                     /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"
     CDS             complement(2485..2901)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(2501..2517)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(2533..2555)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    3102..3301
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             3366..3835
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        3866..3970
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3971..4039,4040..4831)
                     /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(4189..4208)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4991..5579
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5480..5499
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5733..5750
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 caactatggc catttaatgt aaatacttaa gaaaaaaaac caaattaatt ttgatacatg
       61 ctgcatgtga aagacccccg ctgacgggta gtcaatcact cagaggagac cctcccaagg
      121 aacagcgaga ccacaagtcg gatgcaactg caagagggtt tattggatac acgggtaccc
      181 gggcgactca gtcaatcgga ggactggcgc cccgagtgag gggttgtggg ctcttttatt
      241 gagctcgggg agcagaagcg cgcgaacaga agcgagaagc gaactgattg gttagttcaa
      301 ataaggcaca gggtcatttc aggtccttgg ggcaccctgg aaacatctga tggttctcta
      361 gaaactgctg agggctggac cgcatctggg gaccatctgt tcttggccct gagccggggc
      421 aggaactgct taccacagat atcctgtttg gcccatattc agctgttcca tctgttcttg
      481 gccctgagcc ggggcaggaa ctgcttacca cagatatcct gtttggccca tattcagctg
      541 ttccatctgt tcctgacctt gatctgaact tctctattct cagttatgta tttttccatg
      601 ccttgcaaaa tggcgttact taagctagct tgccaaacct acaggtgggg tctttcattc
      661 cccccttttt ctggagacta aataaaatct tttattttat cgatgcatgg ggtcgtgcgc
      721 tcctttcggt cgggcgctgc gggtcgtggg gcgggcgtca ggcaccgggc ttgcgggtca
      781 tgcaccaggt gcgcggtcct tcgggcacct cgacgtcggc ggtgacggtg aagccgagcc
      841 gctcgtagaa ggggaggttg cggggcgcgg aggtctccag gaaggcgggc accccggcgc
      901 gctcggccgc ctccactccg gggagcacga cggcgctgcc cagacccttg ccctggtggt
      961 cgggcgagac gccgacggtg gccaggaacc acgcgggctc cttgggccgg tgcggcgcca
     1021 ggaggccttc catctgttgc tgcgcggcca gccgggaacc gctcaactcg gccatgcgcg
     1081 ggccgatctc ggcgaacacc gcccccgctt cgacgctctc cggcgtggtc cagaccgcca
     1141 ccgcggcgcc gtcgtccgcg acccacacct tgccgatgtc gagcccgacg cgcgtgagga
     1201 agagttcttg cagctcggtg acccgctcga tgtggcggtc cggatcgacg gtgtggcgcg
     1261 tggcggggta gtcggcgaac gcggcggcga gggtgcgtac ggccctgggg acgtcgtcgc
     1321 gggtggcgag gcgcaccgtg ggcttgtact cggtcatggt aagctttttg caaaagccta
     1381 ggcctccaaa aaagcctcct cactacttct ggaatagctc agaggccgag gcggcctcgg
     1441 cctctgcata aataaaaaaa attagtcagc catggggcgg agaatgggcg gaactgggcg
     1501 gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat
     1561 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg
     1621 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     1681 accctaactg acacacattc cacagggtcg accactgtgc tggcgaattc ctacagcttg
     1741 gggggtgact gggctccagc agcccttcct tccttccttg cttccgtcct tccttcctcc
     1801 tccttccgtc tgcacctcct tcctgcatcc ggcacctcca tgtcctgagc ttgtgctgcg
     1861 tcaggagagc acacacttgc agctcatgca gccggggcca ctctcatcag gagggttcag
     1921 cttccgcagc ttgtgctgcc ggatctcacg caccaacgtg tagaaggcat cctccactcc
     1981 ctgccgggtc ttggccgagg tctcgatgta ggggatgccg tagcttcggg cgaggtcctg
     2041 agcctgccga gattccacag tgcgtgcagc caggtcacac ttgttcccca ccagcaccat
     2101 gggcacgtca tccgagtcct tcacccgttt gatctgctcc ctgtactggt ggatgtcctc
     2161 aaaagacttg gtgttgttga tggcaaacac acacaggaag ccctccccgg tgcgcatgta
     2221 ctggtcccgc atggcgctgt actcctcctg gccggcggta tccaggatgt ccaacaggca
     2281 cgtctcccca tcaatgacca cctgcttccg gtaggaatcc tctatagtgg ggtcgtattc
     2341 gtccacaaaa tggttctgga tcagctggat ggtcagcgca ctcttgccca caccgacggc
     2401 gcccaccacc accagcttat attccgtcat cgctcggccc cggccctggc cccccaagct
     2461 gtaccaccac actgggatcc ggccggcgcc tagagaagga gtgagggctg gataaaggga
     2521 ggatcgaggc ggggtcgaac gaggaggttc aagggggaga gacggggcgg atggaggaag
     2581 aggaggcgga ggcttagggt gtacaaaggg cttgacccag ggaggggggt caaaagccaa
     2641 ggcttcccag gtcacgatgt aggggacctg gtctgggtgt ccatgcgggc caggtgaaaa
     2701 gaccttgatc ttaacctggg tgatgaggtc tcggttaaag gtgccgtctc gcggccatcc
     2761 gacgttaaag gttggccatt ctgcagagca gaaggtaacc caacgtctct tcttgacatc
     2821 taccgactgg ttgtgagcga tccgctcgac atctttccag tgacctaagg tcaaacttaa
     2881 gggagtggta acagtctggc cctaattttc agacaaatac agaaacacag tcagacagag
     2941 acaacacaga acgatgctgc agcgctgcag cagacaagac gcgcggcttc ggttccaaac
     3001 cgaaagcaaa aattcagacg gaggcgggaa ctgttttagg ttctcgtctc ctaccagaac
     3061 cacatatcct gacggggtcg gattccacat cgactccctt cctcaggtcg ggccacaaaa
     3121 acggccccca aagtccctgg gacgtctccc agggttgcgg ccgggtgttc agaactcgtc
     3181 agttccacca cgggtccgcc agatacagag ctagttagct aactagtacc gacgcaggcg
     3241 cataaaatca gtcatagaca ctagacaatc ggacagacac agataagttg ctggccagct
     3301 tgcctcccgg tggtgggtcg gtggtccctg ggcaggggtc tcccgatccc ggacgagccc
     3361 ccaaatgaaa gacccccgct gacgggtagt caatcactca gaggagaccc tcccaaggaa
     3421 cagcgagacc acaagtcgga tgcaactgca agagggttta ttggatacac gggtacccgg
     3481 gcgactcagt caatcggagg actggcgccc cgagtgaggg gttgtgggct cttttattga
     3541 gctcggggag cagaagcgcg cgaacagaag cgagaagcga actgattggt tagttcaaat
     3601 aaggcacagg gtcatttcag gtccttgggg caccctggaa acatctgatg gttctctaga
     3661 aactgctgag ggctggaccg catctgggga ccatctgttc ttggccctga gccggggcag
     3721 gaactgctta ccacagatat cctgtttggc ccatattcag ctgttccatc tgttcttggc
     3781 cctgagccgg ggcaggaact gcttaccaca gatatcctgt ttggcccata ttcaggctgc
     3841 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3901 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3961 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4021 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4081 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4141 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4201 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4261 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     4321 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     4381 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     4441 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     4501 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     4561 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     4621 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     4681 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     4741 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     4801 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     4861 ttagattgat ttgcggccgc aaaaggatct aggtgaagat cctttttgat aatctcatga
     4921 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4981 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5041 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5101 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     5161 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5221 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5281 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5341 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5401 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5461 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5521 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5581 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     5641 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     5701 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     5761 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tg
//
