LOCUS       sequence-219889-        5623 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of a constitutively active form of K-Ras.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5623)
  TITLE     Viral vectors.
REFERENCE   2  (bases 1 to 5623)
  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 K-Ras V12
FEATURES             Location/Qualifiers
     source          1..5623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     98..115
                     /label=L4440
                     /note="L4440 vector, forward primer"
     LTR             245..714
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(935..1534)
                     /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(1018..1038)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1515..1534
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(1544..1873)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1558..1693
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1858..1878
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(1890..2420,2421..2423,2424..2456))
                     /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(2473..2889)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(2489..2505)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(2521..2543)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    3090..3289
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             3354..3823
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        3854..3958
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3959..4027,4028..4819)
                     /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(4177..4196)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4979..5567
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5468..5487
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
       61 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
      121 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcaa
      181 ctatggccat ttaatgtaaa tacttaagaa aaaaaaccaa attaattttg atacatgctg
      241 catgtgaaag acccccgctg acgggtagtc aatcactcag aggagaccct cccaaggaac
      301 agcgagacca caagtcggat gcaactgcaa gagggtttat tggatacacg ggtacccggg
      361 cgactcagtc aatcggagga ctggcgcccc gagtgagggg ttgtgggctc ttttattgag
      421 ctcggggagc agaagcgcgc gaacagaagc gagaagcgaa ctgattggtt agttcaaata
      481 aggcacaggg tcatttcagg tccttggggc accctggaaa catctgatgg ttctctagaa
      541 actgctgagg gctggaccgc atctggggac catctgttct tggccctgag ccggggcagg
      601 aactgcttac cacagatatc ctgtttggcc catattcagc tgttccatct gttcttggcc
      661 ctgagccggg gcaggaactg cttaccacag atatcctgtt tggcccatat tcagctgttc
      721 catctgttcc tgaccttgat ctgaacttct ctattctcag ttatgtattt ttccatgcct
      781 tgcaaaatgg cgttacttaa gctagcttgc caaacctaca ggtggggtct ttcattcccc
      841 cctttttctg gagactaaat aaaatctttt attttatcga tgcatggggt cgtgcgctcc
      901 tttcggtcgg gcgctgcggg tcgtggggcg ggcgtcaggc accgggcttg cgggtcatgc
      961 accaggtgcg cggtccttcg ggcacctcga cgtcggcggt gacggtgaag ccgagccgct
     1021 cgtagaaggg gaggttgcgg ggcgcggagg tctccaggaa ggcgggcacc ccggcgcgct
     1081 cggccgcctc cactccgggg agcacgacgg cgctgcccag acccttgccc tggtggtcgg
     1141 gcgagacgcc gacggtggcc aggaaccacg cgggctcctt gggccggtgc ggcgccagga
     1201 ggccttccat ctgttgctgc gcggccagcc gggaaccgct caactcggcc atgcgcgggc
     1261 cgatctcggc gaacaccgcc cccgcttcga cgctctccgg cgtggtccag accgccaccg
     1321 cggcgccgtc gtccgcgacc cacaccttgc cgatgtcgag cccgacgcgc gtgaggaaga
     1381 gttcttgcag ctcggtgacc cgctcgatgt ggcggtccgg atcgacggtg tggcgcgtgg
     1441 cggggtagtc ggcgaacgcg gcggcgaggg tgcgtacggc cctggggacg tcgtcgcggg
     1501 tggcgaggcg caccgtgggc ttgtactcgg tcatggtaag ctttttgcaa aagcctaggc
     1561 ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct
     1621 ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag
     1681 ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca
     1741 tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact
     1801 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
     1861 ctaactgaca cacattccac agggtcgact tacataatta cacactttgt ctttgacttc
     1921 tttttcttct ttttaccatc tttgctcatc ttttctttat gttttcgaat ttctcgaact
     1981 aatgtataga aggcatcatc aacaccctgt cttgtctttg ctgatgtttc aataaaagga
     2041 attccataac ttcttgctaa gtcctgagcc tgttttgtgt ctactgttct ggaaggcaaa
     2101 tcacatttat ttcctactag gaccataggt acatcttcag agtccttaac tcttttaatt
     2161 tgttctctat aatggtgaat atcttcaaat gatttagtat tatttatggc aaatacacaa
     2221 agaaagccct ccccagtcct catgtactgg tccctcattg cactgtactc ctcttgacct
     2281 gctgtgtcga gaatatccaa gagacaggtt tctccatcaa ttactacttg cttcctgtag
     2341 gaatcctcta ttgttggatc atattcgtcc acaaaatgat tctgaattag ctgtatcgtc
     2401 aaggcactct tgcctacgcc aacagctcca actaccacaa gtttatattc agtcatggtg
     2461 gcggatccgg ccggcgccta gagaaggagt gagggctgga taaagggagg atcgaggcgg
     2521 ggtcgaacga ggaggttcaa gggggagaga cggggcggat ggaggaagag gaggcggagg
     2581 cttagggtgt acaaagggct tgacccaggg aggggggtca aaagccaagg cttcccaggt
     2641 cacgatgtag gggacctggt ctgggtgtcc atgcgggcca ggtgaaaaga ccttgatctt
     2701 aacctgggtg atgaggtctc ggttaaaggt gccgtctcgc ggccatccga cgttaaaggt
     2761 tggccattct gcagagcaga aggtaaccca acgtctcttc ttgacatcta ccgactggtt
     2821 gtgagcgatc cgctcgacat ctttccagtg acctaaggtc aaacttaagg gagtggtaac
     2881 agtctggccc taattttcag acaaatacag aaacacagtc agacagagac aacacagaac
     2941 gatgctgcag cgctgcagca gacaagacgc gcggcttcgg ttccaaaccg aaagcaaaaa
     3001 ttcagacgga ggcgggaact gttttaggtt ctcgtctcct accagaacca catatcctga
     3061 cggggtcgga ttccacatcg actcccttcc tcaggtcggg ccacaaaaac ggcccccaaa
     3121 gtccctggga cgtctcccag ggttgcggcc gggtgttcag aactcgtcag ttccaccacg
     3181 ggtccgccag atacagagct agttagctaa ctagtaccga cgcaggcgca taaaatcagt
     3241 catagacact agacaatcgg acagacacag ataagttgct ggccagcttg cctcccggtg
     3301 gtgggtcggt ggtccctggg caggggtctc ccgatcccgg acgagccccc aaatgaaaga
     3361 cccccgctga cgggtagtca atcactcaga ggagaccctc ccaaggaaca gcgagaccac
     3421 aagtcggatg caactgcaag agggtttatt ggatacacgg gtacccgggc gactcagtca
     3481 atcggaggac tggcgccccg agtgaggggt tgtgggctct tttattgagc tcggggagca
     3541 gaagcgcgcg aacagaagcg agaagcgaac tgattggtta gttcaaataa ggcacagggt
     3601 catttcaggt ccttggggca ccctggaaac atctgatggt tctctagaaa ctgctgaggg
     3661 ctggaccgca tctggggacc atctgttctt ggccctgagc cggggcagga actgcttacc
     3721 acagatatcc tgtttggccc atattcagct gttccatctg ttcttggccc tgagccgggg
     3781 caggaactgc ttaccacaga tatcctgttt ggcccatatt caggctgcag gtggcacttt
     3841 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3901 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     3961 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4021 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     4081 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     4141 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4201 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4261 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4321 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     4381 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4441 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4501 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     4561 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4621 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     4681 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     4741 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4801 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     4861 gcggccgcaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4921 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4981 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5041 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5101 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5161 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5221 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5281 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5341 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5401 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5461 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5521 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5581 cggccttttt acggttcctg gccttttgct ggccttttgc tca
//
