LOCUS       sequence-261155-        7086 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of wt CRAF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7086)
  TITLE     pBABEpuro-CRAF
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7086)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pBABEpuro-CRAF
FEATURES             Location/Qualifiers
     source          1..7086
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             8..477
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    542..741
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             942..1358
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1288..1310
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1326..1342
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     protein_bind    1386..1410
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3355..3379)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3416..3436)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3421..3750
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3601..3736
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3760..4359
                     /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(3760..3779)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4256..4276
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4580..5049
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5179..5196)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5350..5938)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5430..5449)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6098..6889,6890..6958))
                     /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     6721..6740
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6959..7063)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
       61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
      121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
      181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
      241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
      301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
      361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
      421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
      481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
      541 caagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
      601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
      661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
      721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
      781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
      841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt
      901 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
      961 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
     1021 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
     1081 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
     1141 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
     1201 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
     1261 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
     1321 cctcccttta tccagccctc actccttctc taggcgccgg ccggatccca gtgtggtggt
     1381 acatcacaag tttgtacaaa aaagttggca tggagcacat acagggagct tggaagacga
     1441 tcagcaatgg ttttggattc aaagatgccg tgtttgatgg ctccagctgc atctctccta
     1501 caatagttca gcagtttggc tatcagcgcc gggcatcaga tgatggcaaa ctcacagatc
     1561 cttctaagac aagcaacact atccgtgttt tcttgccgaa caagcaaaga acagtggtca
     1621 atgtgcgaaa tggaatgagc ttgcatgact gccttatgaa agcactcaag gtgaggggcc
     1681 tgcaaccaga gtgctgtgca gtgttcagac ttctccacga acacaaaggt aaaaaagcac
     1741 gcttagattg gaatactgat gctgcgtctt tgattggaga agaacttcaa gtagatttcc
     1801 tggatcatgt tcccctcaca acacacaact ttgctcggaa gacgttcctg aagcttgcct
     1861 tctgtgacat ctgtcagaaa ttcctgctca atggatttcg atgtcagact tgtggctaca
     1921 aatttcatga gcactgtagc accaaagtac ctactatgtg tgtggactgg agtaacatca
     1981 gacaactctt attgtttcca aattccacta ttggtgatag tggagtccca gcactacctt
     2041 ctttgactat gcgtcgtatg cgagagtctg tttccaggat gcctgttagt tctcagcaca
     2101 gatattctac acctcacgcc ttcaccttta acacctccag tccctcatct gaaggttccc
     2161 tctcccagag gcagaggtcg acatccacac ctaatgtcca catggtcagc accaccctgc
     2221 ctgtggacag caggatgatt gaggatgcaa ttcgaagtca cagcgaatca gcctcacctt
     2281 cagccctgtc cagtagcccc aacaatctga gcccaacagg ctggtcacag ccgaaaaccc
     2341 ccgtgccagc acaaagagag cgggcaccag tatctgggac ccaggagaaa aacaaaatta
     2401 ggcctcgtgg acagagagat tcaagctatt attgggaaat agaagccagt gaagtgatgc
     2461 tgtccactcg gattgggtca ggctcttttg gaactgttta taagggtaaa tggcacggag
     2521 atgttgcagt aaagatccta aaggttgtcg acccaacccc agagcaattc caggccttca
     2581 ggaatgaggt ggctgttctg cgcaaaacac ggcatgtgaa cattctgctt ttcatggggt
     2641 acatgacaaa ggacaacctg gcaattgtga cccagtggtg cgagggcagc agcctctaca
     2701 aacacctgca tgtccaggag accaagtttc agatgttcca gctaattgac attgcccggc
     2761 agacggctca gggaatggac tatttgcatg caaagaacat catccataga gacatgaaat
     2821 ccaacaatat atttctccat gaaggcttaa cagtgaaaat tggagatttt ggtttggcaa
     2881 cagtaaagtc acgctggagt ggttctcagc aggttgaaca acctactggc tctgtcctct
     2941 ggatggcccc agaggtgatc cgaatgcagg ataacaaccc attcagtttc cagtcggatg
     3001 tctactccta tggcatcgta ttgtatgaac tgatgacggg ggagcttcct tattctcaca
     3061 tcaacaaccg agatcagatc atcttcatgg tgggccgagg atatgcctcc ccagatctta
     3121 gtaagctata taagaactgc cccaaagcaa tgaagaggct ggtagctgac tgtgtgaaga
     3181 aagtaaagga agagaggcct ctttttcccc agatcctgtc ttccattgag ctgctccaac
     3241 actctctacc gaagatcaac cggagcgctt ccgagccatc cttgcatcgg gcagcccaca
     3301 ctgaggatat caatgcttgc acgctgacca cgtccccgag gctgcctgtc ttctagccaa
     3361 ctttcttgta caaagtggtg atgtaggaat tcgccagcac agtggtcgac cctgtggaat
     3421 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3481 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3541 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3601 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     3661 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     3721 ggcttttttg gaggcctagg cttttgcaaa aagcttacca tgaccgagta caagcccacg
     3781 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
     3841 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
     3901 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     3961 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     4021 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     4081 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     4141 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     4201 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     4261 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     4321 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag
     4381 cgcccgaccg aaaggagcgc acgaccccat gcatcgataa aataaaagat tttatttagt
     4441 ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta
     4501 acgccatttt gcaaggcatg gaaaaataca taactgagaa tagagaagtt cagatcaagg
     4561 tcaggaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc
     4621 tgccccggct cagggccaag aacagatgga acagctgaat atgggccaaa caggatatct
     4681 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc
     4741 agccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa
     4801 tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct
     4861 tctgctcccc gagctcaata aaagagccca caacccctca ctcggggcgc cagtcctccg
     4921 attgactgag tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac
     4981 ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg
     5041 ggtctttcac atgcagcatg tatcaaaatt aatttggttt tttttcttaa gtatttacat
     5101 taaatggcca tagttgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     5161 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     5221 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     5281 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     5341 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5401 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5461 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     5521 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     5581 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     5641 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5701 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5761 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     5821 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5881 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5941 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6001 gattttggtc atgagattat caaaaaggat cttcacctag atccttttgc ggccgcaaat
     6061 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6121 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6181 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6241 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6301 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6361 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6421 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6481 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6541 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6601 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6661 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6721 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6781 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6841 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6901 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6961 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7021 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7081 tgccac
//
