LOCUS       sequence-177061-        7210 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PI(WT): Photo-inhibitable/blue light sensitive; const. active Rac1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7210)
  AUTHORS   Dagliyan O, Tarnawski M, Chu PH, Shirvanyants D, Schlichting I, 
            Dokholyan NV, Hahn KM
  TITLE     Engineering extrinsic disorder to control protein activity in living
            cells.
  JOURNAL   Science. 2016 Dec 16;354(6318):1441-1444.
  PUBMED    27980211
REFERENCE   2  (bases 1 to 7210)
  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: "Science."; 
            date: "2016-12-16"; volume: "354(6318)"; pages: "1441-1444"
COMMENT     Sequence Label: pBabe-PI(WT)-Rac1(Q61L)-mCherry
FEATURES             Location/Qualifiers
     source          1..7210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     295..314
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             385..984
                     /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(385..404)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     881..901
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(1493..1510)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1664..2252)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1744..1763)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2412..3203,3204..3272))
                     /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     3035..3054
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3273..3377)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             3408..3877
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    3942..4141
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             4342..4758
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     4688..4710
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     4726..4742
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     protein_bind    join(4799..4817,4818..4840,4841..4859,4860..4882,4883..4901,
                     4902..4924,4925..4943,4944..4966,4967..4985,4986..5008,
                     5009..5027,5028..5050,5051..5069)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        5102..5140
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5137..5161
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      5256..5265
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5262..5969
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(5407..5425)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5675..5694
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             6147..6575
                     /codon_start=1
                     /gene="NPH1 from Avena sativa"
                     /product="light, oxygen, or voltage domain from phototropin
                     1 of Avena sativa (Herrou and Crosson, 2011)"
                     /label=LOV2
                     /note="photoswitch that undergoes a light-inducible 
                     conformational change"
                     /translation="LATTLERIEKNFVITDPRLPDNPIIFASDSFLQLTEYSREEILGR
                     NCRFLQGPETDRATVRKIRDAIDNQTEVTVQLINYTKSGKKFWNLFHLQPMRDQKGDVQ
                     YFIGVQLDGTEHVRDAAEREGVMLIKKTAENIDEAAKEL"
     protein_bind    7036..7052
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        join(7085..7210,1..362)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7120..7143)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt
       61 tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca
      121 ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca
      181 gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt
      241 ccgggggcgg gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct
      301 gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt
      361 cgacctgcag ccaataagct taccatgacc gagtacaagc ccacggtgcg cctcgccacc
      421 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
      481 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
      541 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
      601 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
      661 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
      721 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
      781 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
      841 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
      901 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
      961 atgacccgca agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg
     1021 agcgcacgac cccatgcatc gataaaataa aagattttat ttagtctcca gaaaaagggg
     1081 ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag
     1141 gcatggaaaa atacataact gagaatagag aagttcagat caaggtagga gatccctgag
     1201 cccacaaccc ctcactcggg gcgccagtcc tccgattgac tgagtcgccc gggtacccgt
     1261 gtatccaata aaccctcttg cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg
     1321 gtctcctctg agtgattgac tacccgtcag cgggggtctt tcacatgcag catgtatcaa
     1381 aattaatttg gttttttttc ttaagtattt acattaaatg gccatagttg cattaatgaa
     1441 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     1501 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     1561 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     1621 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     1681 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     1741 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     1801 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     1861 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     1921 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     1981 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     2041 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     2101 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     2161 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     2221 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     2281 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     2341 ggatcttcac ctagatcctt ttgcggccgc aaatcaatct aaagtatata tgagtaaact
     2401 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     2461 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     2521 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     2581 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     2641 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     2701 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     2761 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     2821 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     2881 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     2941 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     3001 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     3061 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     3121 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     3181 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     3241 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     3301 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     3361 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgcagcctg aatatgggcc
     3421 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac
     3481 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     3541 aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat
     3601 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
     3661 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
     3721 acccctcact cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc
     3781 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
     3841 tctgagtgat tgactacccg tcagcggggg tctttcattt gggggctcgt ccgggatcgg
     3901 gagacccctg cccagggacc accgacccac caccgggagg caagctggcc agcaacttat
     3961 ctgtgtctgt ccgattgtct agtgtctatg actgatttta tgcgcctgcg tcggtactag
     4021 ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc
     4081 ggccgcaacc ctgggagacg tcccagggac tttgggggcc gtttttgtgg cccgacctga
     4141 ggaagggagt cgatgtggaa tccgaccccg tcaggatatg tggttctggt aggagacgag
     4201 aacctaaaac agttcccgcc tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc
     4261 gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt
     4321 ctgtatttgt ctgaaaatta gggccagact gttaccactc ccttaagttt gaccttaggt
     4381 cactggaaag atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt
     4441 tgggttacct tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc
     4501 acctttaacc gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat
     4561 ggacacccag accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct
     4621 ccctgggtca agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg
     4681 tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc
     4741 actccttctc taggcgccgg ccggatccac gaggcccttt cgtcgtcgag tttaccactc
     4801 cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa
     4861 aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta
     4921 ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat
     4981 agagaaaagt gaaagtcgag tttaccactc cctatcagtg atagagaaaa gtgaaagtcg
     5041 agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagctcggt acccgggtcg
     5101 aggtaggcgt gtacggtggg aggcctatat aagcagagct cgtttagtga accgtcagat
     5161 cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg accgatccag
     5221 cctccgcggc cccagatccg taaagtgaat tcgccgccac catggtgagc aagggcgagg
     5281 aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg gagggctccg
     5341 tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac gagggcaccc
     5401 agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg gacatcctgt
     5461 cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac atccccgact
     5521 acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg
     5581 gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc atctacaagg
     5641 tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag aagaccatgg
     5701 gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag ggcgagatca
     5761 agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag accacctaca
     5821 aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag ttggacatca
     5881 cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag ggccgccact
     5941 ccaccggcgg catggacgag ctgtacaagt ccggactcag atctgagctc ggatccatgc
     6001 aggccatcaa gtgtgtggtg gtgggagacg gagctgtagg taaaacttgc ctactgatca
     6061 gttacacaac caatgcattt cctggagaat atatccctac tgtctttgac aattattctg
     6121 ccaatgttat ggtagatgga tcaggtttgg ctactacact tgaacgtatt gagaagaact
     6181 ttgtcattac tgacccaaga ttgccagata atcccattat attcgcgtcc gatagtttct
     6241 tgcagttgac agaatatagc cgtgaagaaa ttttgggaag aaactgcagg tttctacaag
     6301 gtcctgaaac tgatcgcgcg acagtgagaa aaattagaga tgccatagat aaccaaacag
     6361 aggtcactgt tcagctgatt aattatacaa agagtggtaa aaagttctgg aacctctttc
     6421 acttgcagcc tatgcgagat cagaagggag atgtccagta ctttattggg gttcagttgg
     6481 atggaactga gcatgtccga gatgctgccg agagagaggg agtcatgctg attaagaaaa
     6541 ctgcagaaaa tattgatgag gcggcaaaag aacttggcag tggtggaaaa ccggtgaatc
     6601 tgggcttatg ggatacagct ggactagaag attatgacag attacgcccc ctatcctatc
     6661 cgcaaacaga tgtgttctta atttgctttt cccttgtgag tcctgcatca tttgaaaatg
     6721 tccgtgcaaa gtggtatcct gaggtgcggc accactgtcc caacactccc atcatcctag
     6781 tgggaactaa acttgatctt agggatgata aagacacgat cgagaaactg aaggagaaga
     6841 agctgactcc catcacctat ccgcagggtc tagccatggc taaggagatt ggtgccgtaa
     6901 aatacctgga gtgctcggcg ctcacacagc gaggcctcaa gacagtgttt gacgaagcga
     6961 tccgagcagt cctctgcccg cctcccgtga agaagaggaa gagaaaatgc ctgctgttgt
     7021 aactcgagtg tggaattgtg agcggataac aatttgtcga gggcccctgc aggtcaattc
     7081 taccgggtag gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg
     7141 ctgggcactt ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt
     7201 aggcgccaac
//
