LOCUS       sequence-177112-        7208 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PI(WT): Photo-inhibitable/blue light sensitive Rac1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7208)
  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 7208)
  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(WT)-mCherry
FEATURES             Location/Qualifiers
     source          1..7208
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     315..338
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     protein_bind    406..422
                     /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)."
     CDS             complement(883..1311)
                     /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"
     CDS             complement(1489..2196)
                     /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(1764..1783)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2033..2051
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      2193..2202
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2297..2321)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2318..2356)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    join(2389..2407,2408..2430,2431..2449,2450..2472,2473..2491,
                     2492..2514,2515..2533,2534..2556,2557..2575,2576..2598,
                     2599..2617,2618..2640,2641..2659)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     CDS             complement(2700..3116)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(2716..2732)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(2748..2770)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    3317..3516
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             3581..4050
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     promoter        4081..4185
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4186..4254,4255..5046)
                     /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     complement(4404..4423)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5206..5794
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5695..5714
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5948..5965
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(6474..7073)
                     /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(6557..6577)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     7054..7073
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(join(7096..7208,1..373))
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7144..7163)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
ORIGIN
        1 cccccggacc caccccttcc cagcctctga gcccagaaag cgaaggagca aagctgctat
       61 tggccgctgc cccaaaggcc tacccgcttc cattgctcag cggtgctgtc catctgcacg
      121 agactagtga gacgtgctac ttccatttgt cacgtcctgc acgacgcgag ctgcggggcg
      181 ggggggaact tcctgactag gggaggagta gaaggtggcg cgaaggggcc accaaagaac
      241 ggagccggtt ggcgcctacc ggtggatgtg gaatgtgtgc gaggccagag gccacttgtg
      301 tagcgccaag tgcccagcgg ggctgctaaa gcgcatgctc cagactgcct tgggaaaagc
      361 gcctccccta cccggtagaa ttgacctgca ggggccctcg acaaattgtt atccgctcac
      421 aattccacac tcgagttaca acagcaggca ttttctcttc ctcttcttca cgggaggcgg
      481 gcagaggact gctcggatcg cttcgtcaaa cactgtcttg aggcctcgct gtgtgagcgc
      541 cgagcactcc aggtatttta cggcaccaat ctccttagcc atggctagac cctgcggata
      601 ggtgatggga gtcagcttct tctccttcag tttctcgatc gtgtctttat catccctaag
      661 atcaagttta gttcccacta ggatgatggg agtgttggga cagtggtgcc gcacctcagg
      721 ataccacttt gcacggacat tttcaaatga tgcaggactc acaagggaaa agcaaattaa
      781 gaacacatct gtttgcggat aggatagggg gcgtaatctg tcataatctt cttgtccagc
      841 tgtatcccat aagcccagat tcaccggttt tccaccactg ccaagttctt ttgccgcctc
      901 atcaatattt tctgcagttt tcttaatcag catgactccc tctctctcgg cagcatctcg
      961 gacatgctca gttccatcca actgaacccc aataaagtac tggacatctc ccttctgatc
     1021 tcgcataggc tgcaagtgaa agaggttcca gaacttttta ccactctttg tataattaat
     1081 cagctgaaca gtgacctctg tttggttatc tatggcatct ctaatttttc tcactgtcgc
     1141 gcgatcagtt tcaggacctt gtagaaacct gcagtttctt cccaaaattt cttcacggct
     1201 atattctgtc aactgcaaga aactatcgga cgcgaatata atgggattat ctggcaatct
     1261 tgggtcagta atgacaaagt tcttctcaat acgttcaagt gtagtagcca aacctgatcc
     1321 atctaccata acattggcag aataattgtc aaagacagta gggatatatt ctccaggaaa
     1381 tgcattggtt gtgtaactga tcagtaggca agttttacct acagctccgt ctcccaccac
     1441 cacacacttg atggcctgca tggatccgag ctcagatctg agtccggact tgtacagctc
     1501 gtccatgccg ccggtggagt ggcggccctc ggcgcgttcg tactgttcca cgatggtgta
     1561 gtcctcgttg tgggaggtga tgtccaactt gatgttgacg ttgtaggcgc cgggcagctg
     1621 cacgggcttc ttggccttgt aggtggtctt gacctcagcg tcgtagtggc cgccgtcctt
     1681 cagcttcagc ctctgcttga tctcgccctt cagggcgccg tcctcggggt acatccgctc
     1741 ggaggaggcc tcccagccca tggtcttctt ctgcattacg gggccgtcgg aggggaagtt
     1801 ggtgccgcgc agcttcacct tgtagatgaa ctcgccgtcc tgcagggagg agtcctgggt
     1861 cacggtcacc acgccgccgt cctcgaagtt catcacgcgc tcccacttga agccctcggg
     1921 gaaggacagc ttcaagtagt cggggatgtc ggcggggtgc ttcacgtagg ccttggagcc
     1981 gtacatgaac tgaggggaca ggatgtccca ggcgaagggc agggggccac ccttggtcac
     2041 cttcagcttg gcggtctggg tgccctcgta ggggcggccc tcgccctcgc cctcgatctc
     2101 gaactcgtgg ccgttcacgg agccctccat gtgcaccttg aagcgcatga actccttgat
     2161 gatggccatg ttatcctcct cgcccttgct caccatggtg gcggcgaatt cactttacgg
     2221 atctggggcc gcggaggctg gatcggtccc ggtgtcttct atggaggtca aaacagcgtg
     2281 gatggcgtct ccaggcgatc tgacggttca ctaaacgagc tctgcttata taggcctccc
     2341 accgtacacg cctacctcga cccgggtacc gagctcgact ttcacttttc tctatcactg
     2401 atagggagtg gtaaactcga ctttcacttt tctctatcac tgatagggag tggtaaactc
     2461 gactttcact tttctctatc actgataggg agtggtaaac tcgactttca cttttctcta
     2521 tcactgatag ggagtggtaa actcgacttt cacttttctc tatcactgat agggagtggt
     2581 aaactcgact ttcacttttc tctatcactg atagggagtg gtaaactcga ctttcacttt
     2641 tctctatcac tgatagggag tggtaaactc gacgacgaaa gggcctcgtg gatccggccg
     2701 gcgcctagag aaggagtgag ggctggataa agggaggatc gaggcggggt cgaacgagga
     2761 ggttcaaggg ggagagacgg ggcggatgga ggaagaggag gcggaggctt agggtgtaca
     2821 aagggcttga cccagggagg ggggtcaaaa gccaaggctt cccaggtcac gatgtagggg
     2881 acctggtctg ggtgtccatg cgggccaggt gaaaagacct tgatcttaac ctgggtgatg
     2941 aggtctcggt taaaggtgcc gtctcgcggc catccgacgt taaaggttgg ccattctgca
     3001 gagcagaagg taacccaacg tctcttcttg acatctaccg actggttgtg agcgatccgc
     3061 tcgacatctt tccagtgacc taaggtcaaa cttaagggag tggtaacagt ctggccctaa
     3121 ttttcagaca aatacagaaa cacagtcaga cagagacaac acagaacgat gctgcagcgc
     3181 tgcagcagac aagacgcgcg gcttcggttc caaaccgaaa gcaaaaattc agacggaggc
     3241 gggaactgtt ttaggttctc gtctcctacc agaaccacat atcctgacgg ggtcggattc
     3301 cacatcgact cccttcctca ggtcgggcca caaaaacggc ccccaaagtc cctgggacgt
     3361 ctcccagggt tgcggccggg tgttcagaac tcgtcagttc caccacgggt ccgccagata
     3421 cagagctagt tagctaacta gtaccgacgc aggcgcataa aatcagtcat agacactaga
     3481 caatcggaca gacacagata agttgctggc cagcttgcct cccggtggtg ggtcggtggt
     3541 ccctgggcag gggtctcccg atcccggacg agcccccaaa tgaaagaccc ccgctgacgg
     3601 gtagtcaatc actcagagga gaccctccca aggaacagcg agaccacaag tcggatgcaa
     3661 ctgcaagagg gtttattgga tacacgggta cccgggcgac tcagtcaatc ggaggactgg
     3721 cgccccgagt gaggggttgt gggctctttt attgagctcg gggagcagaa gcgcgcgaac
     3781 agaagcgaga agcgaactga ttggttagtt caaataaggc acagggtcat ttcaggtcct
     3841 tggggcaccc tggaaacatc tgatggttct ctagaaactg ctgagggctg gaccgcatct
     3901 ggggaccatc tgttcttggc cctgagccgg ggcaggaact gcttaccaca gatatcctgt
     3961 ttggcccata ttcagctgtt ccatctgttc ttggccctga gccggggcag gaactgctta
     4021 ccacagatat cctgtttggc ccatattcag gctgcaggtg gcacttttcg gggaaatgtg
     4081 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4141 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     4201 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     4261 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     4321 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     4381 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     4441 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     4501 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     4561 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     4621 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     4681 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     4741 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     4801 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     4861 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     4921 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     4981 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5041 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttgcg gccgcaaaag
     5101 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5161 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5221 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5281 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5341 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5401 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5461 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5521 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5581 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5641 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5701 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5761 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5821 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     5881 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     5941 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     6001 ccccgcgcgt tggccgattc attaatgcaa ctatggccat ttaatgtaaa tacttaagaa
     6061 aaaaaaccaa attaattttg atacatgctg catgtgaaag acccccgctg acgggtagtc
     6121 aatcactcag aggagaccct cccaaggaac agcgagacca caagtcggat gcaactgcaa
     6181 gagggtttat tggatacacg ggtacccggg cgactcagtc aatcggagga ctggcgcccc
     6241 gagtgagggg ttgtgggctc agggatctcc taccttgatc tgaacttctc tattctcagt
     6301 tatgtatttt tccatgcctt gcaaaatggc gttacttaag ctagcttgcc aaacctacag
     6361 gtggggtctt tcattccccc ctttttctgg agactaaata aaatctttta ttttatcgat
     6421 gcatggggtc gtgcgctcct ttcggtcggg cgctgcgggt cgtggggcgg gcgtcaggca
     6481 ccgggcttgc gggtcatgca ccaggtgcgc ggtccttcgg gcacctcgac gtcggcggtg
     6541 acggtgaagc cgagccgctc gtagaagggg aggttgcggg gcgcggaggt ctccaggaag
     6601 gcgggcaccc cggcgcgctc ggccgcctcc actccgggga gcacgacggc gctgcccaga
     6661 cccttgccct ggtggtcggg cgagacgccg acggtggcca ggaaccacgc gggctccttg
     6721 ggccggtgcg gcgccaggag gccttccatc tgttgctgcg cggccagccg ggaaccgctc
     6781 aactcggcca tgcgcgggcc gatctcggcg aacaccgccc ccgcttcgac gctctccggc
     6841 gtggtccaga ccgccaccgc ggcgccgtcg tccgcgaccc acaccttgcc gatgtcgagc
     6901 ccgacgcgcg tgaggaagag ttcttgcagc tcggtgaccc gctcgatgtg gcggtccgga
     6961 tcgacggtgt ggcgcgtggc ggggtagtcg gcgaacgcgg cggcgagggt gcgtacggcc
     7021 ctggggacgt cgtcgcgggt ggcgaggcgc accgtgggct tgtactcggt catggtaagc
     7081 ttattggctg caggtcgaaa ggcccggaga tgaggaagag gagaacagcg cggcagacgt
     7141 gcgcttttga agcgtgcaga atgccgggcc tccggaggac cttcgggcgc ccgccccgcc
     7201 cctgagcc
//
