LOCUS       sequence-415315-        8423 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses RGG-BcLOV fusion for optogenetic membrane recruitment. 
            Includes an mCherry tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8423)
  AUTHORS   Pal AA, Benman W, Mumford TR, Huang Z, Chow BY, Bugaj LJ
  TITLE     Optogenetic clustering and membrane translocation of the BcLOV4 
            photoreceptor.
  JOURNAL   Proc Natl Acad Sci U S A. 2023 Aug 8;120(32):e2221615120. doi: 
            10.1073/pnas.2221615120. Epub 2023 Aug 1.
  PUBMED    37527339
REFERENCE   2  (bases 1 to 8423)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2023-08-8"; volume: "120(32):e2221615120. 
            doi"; pages: " 10.1073/pnas.2221615120. Epub 2023 Aug 1"
COMMENT     Sequence Label: RGG-BcLOV-mCh
FEATURES             Location/Qualifiers
     source          1..8423
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2224..2934
                     /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(2369..2387)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2637..2656
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(2969..2986)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2975..3199
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3245..3673
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3332..3351)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3542..3563
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3682..3702)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3687..4016
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3867..4002
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3929..3948
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4083..4877
                     /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(4137..4156)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4747..4766
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5051..5172
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5088..5107)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5142..5161
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5221..5237)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5221..5237)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5234..5256)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5245..5261
                     /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(5269..5275,5276..5293,5294..5299))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5314..5335
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5452..5469)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5623..6211)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5703..5722)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6382..7173,7174..7242))
                     /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     7005..7024
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7243..7347)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7422..7441)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        7613..7992
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7993..8196
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8147..8167
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8241..8260
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8241..8259
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gcctcagccg gaggcgacga cagacgcggt ggggccgggg gtggtggcta tcgtagaggt
       61 ggtggaaatt cagggggcgg cggcggagga ggctacgatc gtggatacaa cgacaaccgg
      121 gatgaccgtg acaatcgggg cggctccggc ggttatggcc gggacagaaa ttacgaggat
      181 cgcggttata acgggggtgg tgggggagga ggcaatcgcg gctacaataa taaccgcggt
      241 gggggtggag gcggctataa tcggcaggac cgtggagacg gcggttctag taatttctcg
      301 cggggagggt ataacaacag agatgagggc tcagacaaca gaggctcagg cagatcatac
      361 aacaacgatc ggcgcgacaa tggtggtgac gggggtagcg gcagcggtag catggccaca
      421 gacgcaatcg accattacta ccagaacaga gacggccccc cttccaggag ctttagcgac
      481 gaatacgact catacaataa gacagcccct ctgccacctc tgagcgcctg cccagagcgc
      541 gcccagaccc ctaacagcaa gagaatcccc atcacaccca attccgccaa gagcacctcc
      601 gagtctcctc ggagaatcac attctctcgc gacgcaggca gctccaccaa gggcagcacc
      661 cagacaagcc catccaacca gggctccgtg tctagccgga atggctctaa cggaaatggc
      721 ctgggcctgt cctctgccgg caacttcgcc gatttcttta gctccgaggt gttttatatc
      781 gtgctgcaca atcccaccac agcccacaga ttcctgaagt tttgccagag cagggcctgt
      841 tccgagaacc tggagtttct gcagaagatc gacgagtaca ataggctgat cgatgaggcc
      901 acacgcctgc tgtctaccat ctacagcaca tacgtgagca gcgaggcccc taagcagatc
      961 aacatctcta atagccacgc caggcgcctg agccacgaca tcaagggagc aacccacaac
     1021 gtgctgcccg gcctggagga tatcttcaat ggctcccagg agcagatcga gaagctgctg
     1081 gcctctgacc tgtatcctag atttgtgaag caccaggtga ccgcatccgc cacaatggcc
     1141 ctggcaaacg acaaggagag gtacccaggc ctgggcgatt gcttctgtct gacagaccca
     1201 cggagagccg ataatcccat cgtgttcgca agcgacggat tcgtgagcgt gaccggctat
     1261 tcccgctctg atatcatccc aaggaactgc cgcttcctgc agggcagctt taccgaccgg
     1321 caggccacaa agaggctgcg cacctccatc gagaatcgcg aggagacagt ggagctgctg
     1381 ctgaactatc ggaagaatgg cgatcccttt tggaacctgc tgtacgtgtc ccctctgctg
     1441 gacggcaatg gcgatgtgag attctttctg ggcggccaga tcaactgttc taccacaatc
     1501 cacagccgga ccgacgtgct gagaatcctg agcctgaacg acgaggatac aggctccttc
     1561 atcgagggct ctaataagac ccccagcgtg cggtccaagg agtctaacaa tgcctccaac
     1621 tatggcaaga catccttctt taagtctttt aagaagtaca aggccaatac cgtggagatc
     1681 agagacgagg ccggcatgga gggcgagctg atcgatagga tcggcaagct gcagttcaag
     1741 acacaggtgg aggagtttta caccgcctat tctaagtacc tggtgctgag ctatcagcca
     1801 cagtcccaga acctgatcgt gaagtacttc agccccggca tcgtggatat gctgaacctg
     1861 aatatgccta atggcctggt ggccccaatc gtggacaagg atatcttcaa ggtgctgaca
     1921 gagcactccc cttcctctgt gccacggacc tttaagaacg tggtgaggga aggcatcaaa
     1981 gcaggcaggg ccgtgtctgt ggagacaggc ctgctgacag gcatcgagga gatcaagaag
     2041 tctcagaaca tcttcagcgg caccacagcc aatcgggaga gaggctggaa gaaggccgag
     2101 gagaagtacg tgtgccactt taccccctgt aaggatgaga tgggcagggt cggctgggtg
     2161 gtgctgactg tcgctcctaa actggaaaga tttgaattcg gtggtggatc aggtggagga
     2221 tctatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
     2281 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     2341 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
     2401 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
     2461 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     2521 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     2581 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     2641 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     2701 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     2761 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     2821 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     2881 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaatgatct
     2941 agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     3001 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     3061 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3121 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     3181 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     3241 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3301 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3361 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3421 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3481 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3541 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3601 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3661 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3721 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3781 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3841 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3901 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     3961 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     4021 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     4081 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4141 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4201 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4261 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4321 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4381 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4441 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4501 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4561 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     4621 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4681 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4741 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4801 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4861 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     4921 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4981 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5041 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     5101 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5161 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     5221 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5281 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5341 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5401 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5461 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5521 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5581 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5641 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5701 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5761 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5821 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5881 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5941 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6001 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6061 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6121 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6181 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6241 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6301 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6361 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6421 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6481 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6541 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6601 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6661 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6721 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6781 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6841 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6901 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6961 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     7021 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     7081 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7141 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7201 aaaaaaggga ataagggcga catggaaatg ttgaatactc atactcttcc tttttcaata
     7261 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     7321 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     7381 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
     7441 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     7501 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     7561 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga
     7621 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     7681 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     7741 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     7801 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     7861 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     7921 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     7981 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     8041 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     8101 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     8161 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact
     8221 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc tggctagcgt
     8281 ttaaacttaa gcttggtacc gagctcggat ccatggaatc gaatcaatct aacaacggtg
     8341 gatcgggaaa cgcagcattg aatcgtgggg gccgttacgt accaccacac ttgagaggcg
     8401 gagatggtgg tgccgccgct gct
//
