LOCUS       sequence-217979-        7513 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral expression of BRAF V600E (the V600E mutation has been 
            found in cancer patients, confers increased kinase activity, and is 
            transforming).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7513)
  AUTHORS   Boehm JS, Zhao JJ, Yao J, Kim SY, Firestein R, Dunn IF, Sjostrom SK,
            Garraway LA, Weremowicz S, Richardson AL, Greulich H, Stewart CJ, 
            Mulvey LA, Shen RR, Ambrogio L, Hirozane-Kishikawa T, Hill DE, Vidal
            M, Meyerson M, Grenier JK, Hinkle G, Root DE, Roberts TM, Lander ES,
            Polyak K, Hahn WC
  TITLE     Integrative Genomic Approaches Identify IKBKE as a Breast Cancer 
            Oncogene.
  JOURNAL   Cell. 2007 Jun 15. 129(6):1065-1079.
  PUBMED    17574021
REFERENCE   2  (bases 1 to 7513)
  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. 2007 
            Jun 15. 129(6):1065-1079."
COMMENT     Sequence Label: pBabe-Puro-BRAF-V600E
FEATURES             Location/Qualifiers
     source          1..7513
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             10..479
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    544..743
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             944..1360
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1290..1312
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1328..1344
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(1401..1417)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1402..1418)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             3756..3785
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3801..3818
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3845..3865)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3850..4179
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4030..4165
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4189..4788
                     /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(4189..4208)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4685..4705
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5009..5478
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5608..5625)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5779..6367)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5859..5878)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6527..7318,7319..7387))
                     /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     7150..7169
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7388..7492)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acctgcagcc tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
       61 cagggccaag aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca
      121 gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc
      181 agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg
      241 ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc
      301 gagctcaata aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag
      361 tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc
      421 gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcat
      481 ttgggggctc gtccgggatc gggagacccc tgcccaggga ccaccgaccc accaccggga
      541 ggcaagctgg ccagcaactt atctgtgtct gtccgattgt ctagtgtcta tgactgattt
      601 tatgcgcctg cgtcggtact agttagctaa ctagctctgt atctggcgga cccgtggtgg
      661 aactgacgag ttctgaacac ccggccgcaa ccctgggaga cgtcccaggg actttggggg
      721 ccgtttttgt ggcccgacct gaggaaggga gtcgatgtgg aatccgaccc cgtcaggata
      781 tgtggttctg gtaggagacg agaacctaaa acagttcccg cctccgtctg aatttttgct
      841 ttcggtttgg aaccgaagcc gcgcgtcttg tctgctgcag cgctgcagca tcgttctgtg
      901 ttgtctctgt ctgactgtgt ttctgtattt gtctgaaaat tagggccaga ctgttaccac
      961 tcccttaagt ttgaccttag gtcactggaa agatgtcgag cggatcgctc acaaccagtc
     1021 ggtagatgtc aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa
     1081 cgtcggatgg ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa
     1141 ggtcttttca cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga
     1201 agccttggct tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc
     1261 tcctcttcct ccatccgccc cgtctctccc ccttgaacct cctcgttcga ccccgcctcg
     1321 atcctccctt tatccagccc tcactccttc tctaggcgcc ggccggatcc tcgagttaac
     1381 gaattctaca tcaagcttat cgataccgtc gacctcgaat ggcggcgctg agcggtggcg
     1441 gtggtggcgg cgcggagccg ggccaggctc tgttcaacgg ggacatggag cccgaggccg
     1501 gcgccggcgc cggcgccgcg gcctcttcgg ctgcggaccc tgccattccg gaggaggtgt
     1561 ggaatatcaa acaaatgatt aagttgacac aggaacatat agaggcccta ttggacaaat
     1621 ttggtgggga gcataatcca ccatcaatat atctggaggc ctatgaagaa tacaccagca
     1681 agctagatgc actccaacaa agagaacaac agttattgga atctctgggg aacggaactg
     1741 atttttctgt ttctagctct gcatcaatgg ataccgttac atcttcttcc tcttctagcc
     1801 tttcagtgct accttcatct ctttcagttt ttcaaaatcc cacagatgtg gcacggagca
     1861 accccaagtc accacaaaaa cctatcgtta gagtcttcct gcccaacaaa cagaggacag
     1921 tggtacctgc aaggtgtgga gttacagtcc gagacagtct aaagaaagca ctgatgatga
     1981 gaggtctaat cccagagtgc tgtgctgttt acagaattca ggatggagag aagaaaccaa
     2041 ttggttggga cactgatatt tcctggctta ctggagaaga attgcatgtg gaagtgttgg
     2101 agaatgttcc acttacaaca cacaactttg tacgaaaaac gtttttcacc ttagcatttt
     2161 gtgacttttg tcgaaagctg cttttccagg gtttccgctg tcaaacatgt ggttataaat
     2221 ttcaccagcg ttgtagtaca gaagttccac tgatgtgtgt taattatgac caacttgatt
     2281 tgctgtttgt ctccaagttc tttgaacacc acccaatacc acaggaagag gcgtccttag
     2341 cagagactgc cctaacatct ggatcatccc cttccgcacc cgcctcggac tctattgggc
     2401 cccaaattct caccagtccg tctccttcaa aatccattcc aattccacag cccttccgac
     2461 cagcagatga agatcatcga aatcaatttg ggcaacgaga ccgatcctca tcagctccca
     2521 atgtgcatat aaacacaata gaacctgtca atattgatga cttgattaga gaccaaggat
     2581 ttcgtggtga tggaggatca accacaggtt tgtctgctac cccccctgcc tcattacctg
     2641 gctcactaac taacgtgaaa gccttacaga aatctccagg acctcagcga gaaaggaagt
     2701 catcttcatc ctcagaagac aggaatcgaa tgaaaacact tggtagacgg gactcgagtg
     2761 atgattggga gattcctgat gggcagatta cagtgggaca aagaattgga tctggatcat
     2821 ttggaacagt ctacaaggga aagtggcatg gtgatgtggc agtgaaaatg ttgaatgtga
     2881 cagcacctac acctcagcag ttacaagcct tcaaaaatga agtaggagta ctcaggaaaa
     2941 cacgacatgt gaatatccta ctcttcatgg gctattccac aaagccacaa ctggctattg
     3001 ttacccagtg gtgtgagggc tccagcttgt atcaccatct ccatatcatt gagaccaaat
     3061 ttgagatgat caaacttata gatattgcac gacagactgc acagggcatg gattacttac
     3121 acgccaagtc aatcatccac agagacctca agagtaataa tatatttctt catgaagacc
     3181 tcacagtaaa aataggtgat tttggtctag ctacagagaa atctcgatgg agtgggtccc
     3241 atcagtttga acagttgtct ggatccattt tgtggatggc accagaagtc atcagaatgc
     3301 aagataaaaa tccatacagc tttcagtcag atgtatatgc atttggaatt gttctgtatg
     3361 aattgatgac tggacagtta ccttattcaa acatcaacaa cagggaccag ataattttta
     3421 tggtgggacg aggatacctg tctccagatc tcagtaaggt acggagtaac tgtccaaaag
     3481 ccatgaagag attaatggca gagtgcctca aaaagaaaag agatgagaga ccactctttc
     3541 cccaaattct cgcctctatt gagctgctgg cccgctcatt gccaaaaatt caccgcagtg
     3601 catcagaacc ctccttgaat cgggctggtt tccaaacaga ggattttagt ctatatgctt
     3661 gtgcttctcc aaaaacaccc atccaggcag ggggatatgg tgcgtttcct gtccacggaa
     3721 agggcgaatt cgttttcgag tctagagggc ccttcgaaca aaaactcatc tcagaagagg
     3781 atctgaatat gcataccggt catcatcacc atcaccattg agtttgtaga tctgtcgacc
     3841 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3901 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3961 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4021 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4081 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     4141 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttaccat gaccgagtac
     4201 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
     4261 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
     4321 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     4381 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
     4441 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     4501 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     4561 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     4621 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     4681 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     4741 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgacg cccgccccac
     4801 gacccgcagc gcccgaccga aaggagcgca cgaccccatg catcgataaa ataaaagatt
     4861 ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt tggcaagcta
     4921 gcttaagtaa cgccattttg caaggcatgg aaaaatacat aactgagaat agagaagttc
     4981 agatcaaggt caggaacaga tggaacagct gaatatgggc caaacaggat atctgtggta
     5041 agcagttcct gccccggctc agggccaaga acagatggaa cagctgaata tgggccaaac
     5101 aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggtccccag
     5161 atgcggtcca gccctcagca gtttctagag aaccatcaga tgtttccagg gtgccccaag
     5221 gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt
     5281 tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac tcggggcgcc
     5341 agtcctccga ttgactgagt cgcccgggta cccgtgtatc caataaaccc tcttgcagtt
     5401 gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc
     5461 gtcagcgggg gtctttcaca tgcagcatgt atcaaaatta atttggtttt ttttcttaag
     5521 tatttacatt aaatggccat agttgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5581 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5641 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5701 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5761 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5821 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5881 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5941 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6001 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6061 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6121 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6181 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6241 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6301 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6361 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6421 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttgcg
     6481 gccgcaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6541 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6601 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6661 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6721 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6781 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6841 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6901 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6961 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7021 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7081 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7141 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7201 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7261 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7321 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7381 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7441 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7501 cccgaaaagt gcc
//
