LOCUS       sequence-271132-        8498 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses opto-Rac1 (BcLOV4-GGGSx2-WT_Rac1-GGGSx2-mCherry in 
            mammalian cells in the pcDNA3.1 backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8498)
  AUTHORS   Berlew EE, Kuznetsov IA, Yamada K, Bugaj LJ, Chow BY
  TITLE     Optogenetic Rac1 engineered from membrane lipid-binding RGS-LOV for 
            inducible lamellipodia formation.
  JOURNAL   Photochem Photobiol Sci. 2020 Mar 1;19(3):353-361. doi: 
            10.1039/c9pp00434c. Epub 2020 Feb 12.
  PUBMED    32048687
REFERENCE   2  (bases 1 to 8498)
  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: 
            "10.1039/c9pp00434c"; journalName: "Photochem Photobiol Sci"; date: 
            "2020-03-1- 1"; volume: "19"; issue: "3"; pages: "353-361"
COMMENT     Sequence Label: opto-Rac1
FEATURES             Location/Qualifiers
     source          1..8498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3110..3820
                     /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(3255..3273)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3523..3542
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3855..3872)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3861..4085
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4131..4559
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4218..4237)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4428..4449
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4568..4588)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4573..4902
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4753..4888
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4815..4834
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4969..5763
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5023..5042)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5633..5652
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5937..6058
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5974..5993)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6028..6047
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6107..6123)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6107..6123)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6120..6142)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6131..6147
                     /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        complement(join(6155..6161,6162..6179,6180..6185))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6200..6221
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6338..6355)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6509..7097)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6589..6608)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7268..8059,8060..8128))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFHVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7891..7910
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8129..8233)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8308..8327)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc atggccacag acgcaatcga
      721 ccattactac cagaacagag acggcccccc ttccaggagc tttagcgacg aatacgactc
      781 atacaataag acagcccctc tgccacctct gagcgcctgc ccagagcgcg cccagacccc
      841 taacagcaag agaatcccca tcacacccaa ttccgccaag agcacctccg agtctcctcg
      901 gagaatcaca ttctctcgcg acgcaggcag ctccaccaag ggcagcaccc agacaagccc
      961 atccaaccag ggctccgtgt ctagccggaa tggctctaac ggaaatggcc tgggcctgtc
     1021 ctctgccggc aacttcgccg atttctttag ctccgaggtg ttttatatcg tgctgcacaa
     1081 tcccaccaca gcccacagat tcctgaagtt ttgccagagc agggcctgtt ccgagaacct
     1141 ggagtttctg cagaagatcg acgagtacaa taggctgatc gatgaggcca cacgcctgct
     1201 gtctaccatc tacagcacat acgtgagcag cgaggcccct aagcagatca acatctctaa
     1261 tagccacgcc aggcgcctga gccacgacat caagggagca acccacaacg tgctgcccgg
     1321 cctggaggat atcttcaatg gctcccagga gcagatcgag aagctgctgg cctctgacct
     1381 gtatcctaga tttgtgaagc accaggtgac cgcatccgcc acaatggccc tggcaaacga
     1441 caaggagagg tacccaggcc tgggcgattg cttctgtctg acagacccac ggagagccga
     1501 taatcccatc gtgttcgcaa gcgacggatt cgtgagcgtg accggctatt cccgctctga
     1561 tatcatccca aggaactgcc gcttcctgca gggcagcttt accgaccggc aggccacaaa
     1621 gaggctgcgc acctccatcg agaatcgcga ggagacagtg gagctgctgc tgaactatcg
     1681 gaagaatggc gatccctttt ggaacctgct gtacgtgtcc cctctgctgg acggcaatgg
     1741 cgatgtgaga ttctttctgg gcggccagat caactgttct accacaatcc acagccggac
     1801 cgacgtgctg agaatcctga gcctgaacga cgaggataca ggctccttca tcgagggctc
     1861 taataagacc cccagcgtgc ggtccaagga gtctaacaat gcctccaact atggcaagac
     1921 atccttcttt aagtctttta agaagtacaa ggccaatacc gtggagatca gagacgaggc
     1981 cggcatggag ggcgagctga tcgataggat cggcaagctg cagttcaaga cacaggtgga
     2041 ggagttttac accgcctatt ctaagtacct ggtgctgagc tatcagccac agtcccagaa
     2101 cctgatcgtg aagtacttca gccccggcat cgtggatatg ctgaacctga atatgcctaa
     2161 tggcctggtg gccccaatcg tggacaagga tatcttcaag gtgctgacag agcactcccc
     2221 ttcctctgtg ccacggacct ttaagaacgt ggtgagggaa ggcatcaaag caggcagggc
     2281 cgtgtctgtg gagacaggcc tgctgacagg catcgaggag atcaagaagt ctcagaacat
     2341 cttcagcggc accacagcca atcgggagag aggctggaag aaggccgagg agaagtacgt
     2401 gtgccacttt accccctgta aggatgagat gggcagggtc ggctgggtgg tgctgactgt
     2461 cgctcctaaa ctggagagat ttgaattcgg tggtggatca ggtggaggat ctatgcaagc
     2521 cataaagtgc gtggttgtag gagacggtgc agtagggaag acttgtttgc ttatatctta
     2581 caccacaaac gcgtttcctg gcgaatacat ccctacggta tttgacaact attctgctaa
     2641 tgtgatggtg gatgggaagc ctgtaaatct tgggctttgg gataccgcag ggcaggagga
     2701 ctatgataga ttgcggccct tgtcttaccc tcaaacagac gtttttttga tctgtttctc
     2761 tctggtcagt cctgcatcct tcgagaatgt aagggctaag tggtacccag aggttaggca
     2821 tcattgtcca aacacgccca ttatattggt tggaaccaag ttggacctcc gggatgataa
     2881 ggatacaatt gagaagctca aggagaagaa gctgactcct ataacttacc cacagggact
     2941 cgcgatggca aaggaaatcg gagcagttaa atacctcgaa tgcagtgctc tgacacagcg
     3001 gggtctgaag acagtctttg atgaggcgat ccgcgccgtt ttgtgcccgc cgcctgtaaa
     3061 aaagcggaaa cgcaagtgtc tgctcggtgg tggatcaggt ggaggatcta tggtgagcaa
     3121 gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga
     3181 gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga
     3241 gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga
     3301 catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat
     3361 ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt
     3421 cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat
     3481 ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa
     3541 gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg
     3601 cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac
     3661 cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt
     3721 ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg
     3781 ccgccactcc accggcggca tggacgagct gtacaagtaa tgatctagag ggcccgttta
     3841 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     3901 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3961 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     4021 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     4081 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     4141 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4201 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4261 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4321 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     4381 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4441 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4501 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     4561 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4621 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4681 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4741 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4801 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     4861 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     4921 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4981 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     5041 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     5101 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     5161 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     5221 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     5281 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     5341 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     5401 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     5461 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     5521 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     5581 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5641 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5701 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5761 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     5821 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     5881 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     5941 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     6001 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     6061 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     6121 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6181 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6241 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     6301 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6361 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6421 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6481 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6541 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6601 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6661 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6721 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6781 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6841 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6901 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6961 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7021 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     7081 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     7141 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7201 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7261 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     7321 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     7381 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7441 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7501 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7561 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7621 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7681 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7741 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7801 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7861 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7921 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7981 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     8041 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     8101 gggcgacatg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     8161 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     8221 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     8281 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     8341 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     8401 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
     8461 tgcgctgctt cgcgatgtac gggccagata tacgcgtt
//
