LOCUS       sequence-213358-        6747 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Rac1 fused with light-insensitive LOV2 (C450A 
            mutation).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6747)
  AUTHORS   Wu YI, Frey D, Lungu OI, Jaehrig A, Schlichting I, Kuhlman B, Hahn 
            KM
  TITLE     A genetically encoded photoactivatable Rac controls the motility of 
            living cells.
  JOURNAL   Nature. 2009 Sep 3. 461(7260):104-8.
  PUBMED    19693014
REFERENCE   2  (bases 1 to 6747)
  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: "Nature. 
            2009 Sep 3. 461(7260):104-8."
COMMENT     Sequence Label: pTriEx-mCherry-PA-Rac1-C450A
FEATURES             Location/Qualifiers
     source          1..6747
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        102..405
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        407..609
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     560..580
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     812..834
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     896..915
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     957..976
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        957..975
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    976..1000
                     /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        1016..1125
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     CDS             1146..1163
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1170..1874
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1312..1330)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1580..1599
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1881..2309
                     /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
                     NARFLQGPETDRATVRKIRDAIDNQTEVTVQLINYTKSGKKFWNLFHLQPMRDQKGDVQ
                     YFIGVQLDGTEHVRDAAEREGVMLIKKTAENIDEAAKEL"
     CDS             2916..2948
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /label=HSV tag
                     /translation="QPELAPEDPED"
     CDS             2955..2978
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(3068..3087)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3133..3188
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3187..3206)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     terminator      3276..3323
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     3336..4041
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           3424..3796
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(3453..4041)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     protein_bind    complement(4050..4083)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(4251..4838)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4330..4349)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4851..5642,5643..5711))
                     /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     5474..5493
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     misc_recomb     join(5954..6747,1..34)
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             5954..6078
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(6116..6721)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
ORIGIN
        1 acaaactgga aatgtctatc aatatatagt tgctgatggc cggcctatta atagtaatca
       61 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
      121 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      181 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      241 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtccgcc ccctattgac
      301 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt acgggacttt
      361 cctacttggc agtacatcta cgtattagtc atcgctatta ccatgctgat gcggttttgg
      421 cagtacacca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
      481 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
      541 aataaccccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      601 agcagacgtc gtttagtgaa ccgtcagatc actagatgct ttattgcggt agtttatcac
      661 agttaaattg ctaacgccag tctcgaactt aacgtgcaga agttggtcgt gaggcactgg
      721 gcaggtaagt atcgggccct ttgtgcgggg ggagcggctc ggggctgtcc gcggggggac
      781 ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc
      841 tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac
      901 gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tggatcggac cgaaattaat
      961 acgactcact ataggggaat tgtgagcgga taacaattcc ccggagttaa tccgggacct
     1021 ttaattcaac ccaacacaat atattatagt taaataagaa ttattatcaa atcatttgta
     1081 tattaattaa aatactatac tgtaaattac attttattta caatcaaagg agatatacca
     1141 tggcacacca tcaccaccat cacggatctg tgagcaaggg cgaggaggat aacatggcca
     1201 tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt
     1261 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga
     1321 aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt
     1381 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct
     1441 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg
     1501 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca
     1561 ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct
     1621 ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc
     1681 tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg
     1741 tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg
     1801 actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg
     1861 acgagctgta caagggatcc ttggctacta cacttgaacg tattgagaag aactttgtca
     1921 ttactgaccc aagattgcca gataatccca ttatattcgc gtccgatagt ttcttgcagt
     1981 tgacagaata tagccgtgaa gaaattttgg gaagaaacgc aaggtttcta caaggtcctg
     2041 aaactgatcg cgcgacagtg agaaaaatta gagatgccat agataaccaa acagaggtca
     2101 ctgttcagct gattaattat acaaagagtg gtaaaaagtt ctggaacctc tttcacttgc
     2161 agcctatgcg agatcagaag ggagatgtcc agtactttat tggggttcag ttggatggaa
     2221 ctgagcatgt ccgagatgct gccgagagag agggagtcat gctgattaag aaaactgcag
     2281 aaaatattga tgaggcggca aaagaactta tcaagtgtgt ggtggtggga gacggagctg
     2341 taggtaaaac ttgcctactg atcagttaca caaccaatgc atttcctgga gaatatatcc
     2401 ctactgtctt tgacaattat tctgccaatg ttatggtaga tggaaaaccg gtgaatctgg
     2461 gcttatggga tacagctgga ctagaagatt atgacagatt acgcccccta tcctatccgc
     2521 aaacagatgt gttcttaatt tgcttttccc ttgtgagtcc tgcatcattc caccacgtcc
     2581 gtgcaaagtg gtatcctgag gtgcggcacc actgtcccaa cactcccatc atcctagtgg
     2641 gaactaaact tgatcttagg gatgataaag acacgatcga gaaactgaag gagaagaagc
     2701 tgactcccat cacctatccg cagggtctag ccatggctaa ggagattggt gctgtaaaat
     2761 acctggagtg ctcggcgctc acacagcgag gcctcaagac agtgtttgac gaagcgatcc
     2821 gagcagtcct ctgcccgcct cccgtgaaga agaggaagag aaaatgcctg ctgttgtaaa
     2881 agcttgcggc cgcacagctg tatacacgtg caagccagcc agaactcgcc ccggaagacc
     2941 ccgaggatct cgagcaccac catcaccatc accatcacta agtgattaac ctcaggtgca
     3001 ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg
     3061 agatcgatct ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc
     3121 tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt
     3181 gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt
     3241 tggtttagag tttggcaaca tatgcccata tgtaactagc ataacccctt ggggcctcta
     3301 aacgggtctt gaggggtttt ttgctgaaag catgcggagg aaattctcct tgaagtttcc
     3361 ctggtgttca aagtaaagga gtttgcacca gacgcacctc tgttcactgg tccggcgtat
     3421 taaaacacga tacattgtta ttagtacatt tattaagcgc tagattctgt gcgttgttga
     3481 tttacagaca attgttgtac gtattttaat aattcattaa atttataatc tttagggtgg
     3541 tatgttagag cgaaaatcaa atgattttca gcgtctttat atctgaattt aaatattaaa
     3601 tcctcaatag atttgtaaaa taggtttcga ttagtttcaa acaagggttg tttttccgaa
     3661 ccgatggctg gactatctaa tggattttcg ctcaacgcca caaaacttgc caaatcttgt
     3721 agcagcaatc tagctttgtc gatattcgtt tgtgttttgt tttgtaataa aggttcgacg
     3781 tcgttcaaaa tattatgcgc ttttgtattt ctttcatcac tgtcgttagt gtacaattga
     3841 ctcgacgtaa acacgttaaa tagagcttgg acatatttaa catcgggcgt gttagcttta
     3901 ttaggccgat tatcgtcgtc gtcccaaccc tcgtcgttag aagttgcttc cgaagacgat
     3961 tttgccatag ccacacgacg cctattaatt gtgtcggcta acacgtccgc gatcaaattt
     4021 gtagttgagc tttttggaat tgcgatcgca taacttcgta tagcatacat tatacgaagt
     4081 tataagctcg gaacgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4141 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4201 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4261 ctccgccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4321 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4381 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4441 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4501 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4561 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4621 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4681 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4741 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4801 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg ttaccaatgc
     4861 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4921 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     4981 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5041 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5101 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5161 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5221 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5281 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5341 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5401 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5461 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5521 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     5581 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     5641 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     5701 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     5761 atgtccgcgc gtttcctgca tcttttaatc aaatcccaag atgtgtataa acgcgccggt
     5821 atgtacagga agaggtttat actaaactgt tacattgcaa acgtggtttc gtgtgccaag
     5881 tgtgaaaacc gatgtttaat caaggctctg acgcatttct acaaccacga ctccaagtgt
     5941 gtgggtgaag tcatgcatct tttaatcaaa tcccaagatg tgtataaacc accaaactgc
     6001 caaaaaatga aaactgtcga caagctctgt ccgtttgctg gcaactgcaa gggtctcaat
     6061 cctatttgta attattgaat aataaaacaa ttataaatgt caaatttgtt ttttattaac
     6121 gatacaaacc aaacgcaaca agaacatttg tagtattatc tataattgaa aacgcgtagt
     6181 tataatcgct gaggtaatat ttaaaatcat tttcaaatga ttcacagtta atttgcgaca
     6241 atataatttt attttcacat aaactagacg ccttgtcgtc ttcttcttcg tattccttct
     6301 ctttttcatt tttctcttca taaaaattaa catagttatt atcgtatcca tatatgtatc
     6361 tatcgtatag agtaaatttt ttgttgtcat aaatatatat gtctttttta atggggtgta
     6421 tagtaccgct gcgcatagtt tttctgtaat ttacaacagt gctattttct ggtagttctt
     6481 cggagtgtgt tgctttaatt attaaattta tataatcaat gaatttggga tcgtcggttt
     6541 tgtacaatat gttgccggca tagtacgcag cttcttctag ttcaattaca ccatttttta
     6601 gcagcaccgg attaacataa ctttccaaaa tgttgtacga accgttaaac aaaaacagtt
     6661 cacctccctt ttctatacta ttgtctgcga gcagttgttt gttgttaaaa ataacagcca
     6721 ttgtaatgag acgcacaaac taatatc
//
