LOCUS       sequence-253055-        5867 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For mammalian cell expression of P29S mutant murine RAC1 carrying 2x
            Myc epitope tag sequences at the N-terminus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5867)
  AUTHORS   Lionarons DA, Hancock DC, Rana S, East P, Moore C, Murillo MM, 
            Carvalho J, Spencer-Dene B, Herbert E, Stamp G, Damry D, Calado DP, 
            Rosewell I, Fritsch R, Neubig RR, Molina-Arcas M, Downward J
  TITLE     RAC1(P29S) Induces a Mesenchymal Phenotypic Switch via Serum 
            Response Factor to Promote Melanoma Development and Therapy 
            Resistance.
  JOURNAL   Cancer Cell. 2019 Jul 8;36(1):68-83.e9. doi: 
            10.1016/j.ccell.2019.05.015. Epub 2019 Jun 27.
  PUBMED    31257073
REFERENCE   2  (bases 1 to 5867)
  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"; doi: 
            "10.1016/j.ccell.2019.05.015"; journalName: "Cancer Cell"; date: 
            "2019-07-8- 8"; volume: "36"; issue: "1"; pages: "68-83.e9"
COMMENT     Sequence Label: pBABE-hygro-2xMyc-Rac1-P29S
FEATURES             Location/Qualifiers
     source          1..5867
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..470
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    535..734
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             935..1351
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1281..1303
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1319..1335
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1368..1397
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1398..1427
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2057..2077)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2062..2391
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2242..2377
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2465..3490
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             2465..3490
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     primer_bind     4007..4026
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     LTR             4288..4757
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     CDS             complement(join(4872..5663,5664..5732))
                     /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     5495..5514
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5733..5837)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
       61 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
      121 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
      181 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
      241 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
      301 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg
      361 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
      421 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca tttgggggct
      481 cgtccgggat cgggagaccc ctgcccaggg accaccgacc caccaccggg aggcaagctg
      541 gccagcaact tatctgtgtc tgtccgattg tctagtgtct atgactgatt ttatgcgcct
      601 gcgtcggtac tagttagcta actagctctg tatctggcgg acccgtggtg gaactgacga
      661 gttctgaaca cccggccgca accctgggag acgtcccagg gactttgggg gccgtttttg
      721 tggcccgacc tgaggaaggg agtcgatgtg gaatccgacc ccgtcaggat atgtggttct
      781 ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc tttcggtttg
      841 gaaccgaagc cgcgcgtctt gtctgctgca gcgctgcagc atcgttctgt gttgtctctg
      901 tctgactgtg tttctgtatt tgtctgaaaa ttagggccag actgttacca ctcccttaag
      961 tttgacctta ggtcactgga aagatgtcga gcggatcgct cacaaccagt cggtagatgt
     1021 caagaagaga cgttgggtta ccttctgctc tgcagaatgg ccaaccttta acgtcggatg
     1081 gccgcgagac ggcaccttta accgagacct catcacccag gttaagatca aggtcttttc
     1141 acctggcccg catggacacc cagaccaggt cccctacatc gtgacctggg aagccttggc
     1201 ttttgacccc cctccctggg tcaagccctt tgtacaccct aagcctccgc ctcctcttcc
     1261 tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg accccgcctc gatcctccct
     1321 ttatccagcc ctcactcctt ctctaggcgc cggccggatc caccatggag cagaagctga
     1381 tctcagagga ggatctggag cagaagctga tctcagagga ggatctggat atccagcaca
     1441 gtggcggccg ccaggccatc aagtgtgtgg tggtgggaga cggagctgtt ggtaaaacct
     1501 gcctgctcat cagttacacg accaatgcat tcagtggaga gtacatcccc accgtctttg
     1561 acaactattc tgccaatgtt atggtagatg gaaaaccagt gaatctgggc ctatgggaca
     1621 cagctggaca agaagattat gacagattgc gtcccctctc ctacccgcag acagacgtgt
     1681 tcttaatttg cttttccctt gtgagtcctg catcatttga aaatgtccgt gcaaagtggt
     1741 atcctgaagt gcgacaccac tgtcccaata ctcctatcat cctcgtgggg acgaagcttg
     1801 atcttaggga tgataaggac accattgaga agctgaagga gaagaagctg actcccatca
     1861 cctacccgca ggggctggcc atggcgaaag agatcggtgc tgtcaaatac ctggagtgct
     1921 cagctctcac acagcgagga ctcaagacag tgtttgacga agctatccga gcggttctct
     1981 gtccccctcc tgtcaagaag aggaagagaa aatgcctgct gttgtaagaa ttcgccagca
     2041 cagtggtcga ccctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     2101 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     2161 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     2221 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     2281 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     2341 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagcttgcc
     2401 agctggggcg ccctctggta aggttgggaa gccctgcaga tctgatcccg gggggcaatg
     2461 agatatgaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt
     2521 cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt
     2581 cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa
     2641 agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga
     2701 cattggggaa tttagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac
     2761 gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggccat
     2821 ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca
     2881 aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt
     2941 gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga
     3001 tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt
     3061 cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga
     3121 ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt
     3181 ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc
     3241 gccgcggctc cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt
     3301 tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc
     3361 cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga
     3421 tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc
     3481 aaaggaatag agtagatgcc gaccgggatc tatcgatgcg gcgggtgtgg tggttacgcg
     3541 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3601 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     3661 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     3721 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3781 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3841 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3901 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3961 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4021 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4081 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc atcgataaaa taaaagattt
     4141 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
     4201 cttaagtaac gccattttgc aaggcatgga aaaatacata actgagaata gagaagttca
     4261 gatcaaggtc aggaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa
     4321 gcagttcctg ccccggctca gggccaagaa cagatggaac agctgaatat gggccaaaca
     4381 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga
     4441 tgcggtccag ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg
     4501 acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt
     4561 cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact cggggcgcca
     4621 gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg
     4681 catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg
     4741 tcagcggggg tctttcacat gcagcatgta tcaaaattaa tttggttttt tttcttaagt
     4801 atttacatta aatggccata gtgcggccgc aaatcaatct aaagtatata tgagtaaact
     4861 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4921 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4981 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5041 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5101 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5161 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5221 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5281 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5341 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5401 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5461 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5521 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5581 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5641 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5701 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5761 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5821 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgcagc
//
