LOCUS       sequence-154443-        7360 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 7360)
  AUTHORS   Hao H, Muniz-Medina VM, Mehta H, Thomas NE, Khazak V, Der CJ, 
            Shields JM
  TITLE     Context-dependent roles of mutant B-Raf signaling in melanoma and 
            colorectal carcinoma cell growth.
  JOURNAL   Mol Cancer Ther. 2007 Aug . 6(8):2220-9.
  PUBMED    17699719
REFERENCE   2  (bases 1 to 7360)
  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: "Mol Cancer 
            Ther. 2007 Aug . 6(8):2220-9."
COMMENT     Sequence Label: pBabe B-Raf V600E
FEATURES             Location/Qualifiers
     source          1..7360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(33..824,825..893))
                     /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     656..675
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(894..998)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             1029..1498
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    1563..1762
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1963..2379
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     2309..2331
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     2347..2363
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(4711..4731)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4716..5045
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4896..5031
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5055..5654
                     /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(5055..5074)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5551..5571
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5875..6344
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6474..6491)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6645..7233)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6725..6744)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
       61 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
      121 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
      181 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
      241 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
      301 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
      361 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
      421 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
      481 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
      541 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
      601 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
      661 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
      721 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
      781 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
      841 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
      901 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
      961 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     1021 cctgcagcct gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc
     1081 agggccaaga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
     1141 ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtcca gccctcagca
     1201 gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
     1261 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
     1321 agctcaataa aagagcccac aacccctcac tcggggcgcc agtcctccga ttgactgagt
     1381 cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg
     1441 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatt
     1501 tgggggctcg tccgggatcg ggagacccct gcccagggac caccgaccca ccaccgggag
     1561 gcaagctggc cagcaactta tctgtgtctg tccgattgtc tagtgtctat gactgatttt
     1621 atgcgcctgc gtcggtacta gttagctaac tagctctgta tctggcggac ccgtggtgga
     1681 actgacgagt tctgaacacc cggccgcaac cctgggagac gtcccaggga ctttgggggc
     1741 cgtttttgtg gcccgacctg aggaagggag tcgatgtgga atccgacccc gtcaggatat
     1801 gtggttctgg taggagacga gaacctaaaa cagttcccgc ctccgtctga atttttgctt
     1861 tcggtttgga accgaagccg cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt
     1921 tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt agggccagac tgttaccact
     1981 cccttaagtt tgaccttagg tcactggaaa gatgtcgagc ggatcgctca caaccagtcg
     2041 gtagatgtca agaagagacg ttgggttacc ttctgctctg cagaatggcc aacctttaac
     2101 gtcggatggc cgcgagacgg cacctttaac cgagacctca tcacccaggt taagatcaag
     2161 gtcttttcac ctggcccgca tggacaccca gaccaggtcc cctacatcgt gacctgggaa
     2221 gccttggctt ttgacccccc tccctgggtc aagccctttg tacaccctaa gcctccgcct
     2281 cctcttcctc catccgcccc gtctctcccc cttgaacctc ctcgttcgac cccgcctcga
     2341 tcctcccttt atccagccct cactccttct ctaggcgccg gccggatctc gactaggaat
     2401 ggcggcgctg agcggtggcg gtggtggcgg cgcggagccg ggccaggctc tgttcaacgg
     2461 ggacatggag cccgaggccg gcgccggcgc cggcgccgcg gcctcttcgg ctgcggaccc
     2521 tgccattccg gaggaggtgt ggaatatcaa acaaatgatt aagttgacac aggaacatat
     2581 agaggcccta ttggacaaat ttggtgggga gcataatcca ccatcaatat atctggaggc
     2641 ctatgaagaa tacaccagca agctagatgc actccaacaa agagaacaac agttattgga
     2701 atctctgggg aacggaactg atttttctgt ttctagctct gcatcaatgg ataccgttac
     2761 atcttcttcc tcttctagcc tttcagtgct accttcatct ctttcagttt ttcaaaatcc
     2821 cacagatgtg gcacggagca accccaagtc accacaaaaa cctatcgtta gagtcttcct
     2881 gcccaacaaa cagaggacag tggtacctgc aaggtgtgga gttacagtcc gagacagtct
     2941 aaagaaagca ctgatgatga gaggtctaat cccagagtgc tgtgctgttt acagaattca
     3001 ggatggagag aagaaaccaa ttggttggga cactgatatt tcctggctta ctggagaaga
     3061 attgcatgtg gaagtgttgg agaatgttcc acttacaaca cacaactttg tacgaaaaac
     3121 gtttttcacc ttagcatttt gtgacttttg tcgaaagctg cttttccagg gtttccgctg
     3181 tcaaacatgt ggttataaat ttcaccagcg ttgtagtaca gaagttccac tgatgtgtgt
     3241 taattatgac caacttgatt tgctgtttgt ctccaagttc tttgaacacc acccaatacc
     3301 acaggaagag gcgtccttag cagagactgc cctaacatct ggatcatccc cttccgcacc
     3361 cgcctcggac tctattgggc cccaaattct caccagtccg tctccttcaa aatccattcc
     3421 aattccacag cccttccgac cagcagatga agatcatcga aatcaatttg ggcaacgaga
     3481 ccgatcctca tcagctccca atgtgcatat aaacacaata gaacctgtca atattgatga
     3541 cttgattaga gaccaaggat ttcgtggtga tggaggatca accacaggtt tgtctgctac
     3601 cccccctgcc tcattacctg gctcactaac taacgtgaaa gccttacaga aatctccagg
     3661 acctcagcga gaaaggaagt catcttcatc ctcagaagac aggaatcgaa tgaaaacact
     3721 tggtagacgg gactcgagtg atgattggga gattcctgat gggcagatta cagtgggaca
     3781 aagaattgga tctggatcat ttggaacagt ctacaaggga aagtggcatg gtgatgtggc
     3841 agtgaaaatg ttgaatgtga cagcacctac acctcagcag ttacaagcct tcaaaaatga
     3901 agtaggagta ctcaggaaaa cacgacatgt gaatatccta ctcttcatgg gctattccac
     3961 aaagccacaa ctggctattg ttacccagtg gtgtgagggc tccagcttgt atcaccatct
     4021 ccatatcatt gagaccaaat ttgagatgat caaacttata gatattgcac gacagactgc
     4081 acagggcatg gattacttac acgccaagtc aatcatccac agagacctca agagtaataa
     4141 tatatttctt catgaagacc tcacagtaaa aataggtgat tttggtctag ctacagagaa
     4201 atctcgatgg agtgggtccc atcagtttga acagttgtct ggatccattt tgtggatggc
     4261 accagaagtc atcagaatgc aagataaaaa tccatacagc tttcagtcag atgtatatgc
     4321 atttggaatt gttctgtatg aattgatgac tggacagtta ccttattcaa acatcaacaa
     4381 cagggaccag ataattttta tggtgggacg aggatacctg tctccagatc tcagtaaggt
     4441 acggagtaac tgtccaaaag ccatgaagag attaatggca gagtgcctca aaaagaaaag
     4501 agatgagaga ccactctttc cccaaattct cgcctctatt gagctgctgg cccgctcatt
     4561 gccaaaaatt caccgcagtg catcagaacc ctccttgaat cgggctggtt tccaaacaga
     4621 ggattttagt ctatatgctt gtgcttctcc aaaaacaccc atccaggcag ggggatatgg
     4681 tgcgtttcct gtccactgag tcgaccctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4741 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4801 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4861 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4921 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4981 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     5041 gcaaaaagct taccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
     5101 tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
     5161 ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
     5221 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
     5281 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
     5341 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
     5401 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg
     5461 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
     5521 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
     5581 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
     5641 agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac
     5701 cccatgcatc gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag
     5761 accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa
     5821 atacataact gagaatagag aagttcagat caaggtcagg aacagatgga acagctgaat
     5881 atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag
     5941 atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
     6001 cagggccaag aacagatggt ccccagatgc ggtccagccc tcagcagttt ctagagaacc
     6061 atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa
     6121 ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga
     6181 gcccacaacc cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc cgggtacccg
     6241 tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag
     6301 ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcacatgca gcatgtatca
     6361 aaattaattt ggtttttttt cttaagtatt tacattaaat ggccatagtt gcattaatga
     6421 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     6481 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     6541 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     6601 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6661 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6721 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6781 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     6841 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6901 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6961 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     7021 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     7081 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     7141 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     7201 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     7261 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     7321 aggatcttca cctagatcct tttgcggccg caaatcaatc
//
