LOCUS       sequence-415156-        8408 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Fus-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 8408)
  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 8408)
  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: Fus-BcLOV-mCh
FEATURES             Location/Qualifiers
     source          1..8408
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             134..928
                     /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(188..207)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     798..817
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1102..1223
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1139..1158)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1193..1212
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1272..1288)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1272..1288)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1285..1307)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1296..1312
                     /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(1320..1326,1327..1344,1345..1350))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1365..1386
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1503..1520)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1674..2262)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1754..1773)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2433..3224,3225..3293))
                     /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     3056..3075
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3294..3398)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3473..3492)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3664..4043
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4044..4247
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4198..4218
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4292..4311
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4292..4310
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4744..4755
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             6683..7393
                     /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(6828..6846)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     7096..7115
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(7428..7445)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7434..7658
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      7704..8132
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7791..7810)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8001..8022
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8141..8161)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(8146..8408,1..67)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(8326..8408,1..53)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8388..8407
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
       61 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
      121 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
      181 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
      241 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
      301 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
      361 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
      421 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
      481 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
      541 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
      601 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
      661 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
      721 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
      781 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
      841 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
      901 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
      961 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     1021 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     1081 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     1141 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     1201 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     1261 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     1321 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     1381 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     1441 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     1501 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     1561 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     1621 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     1681 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     1741 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     1801 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     1861 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     1921 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     1981 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2041 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2101 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2161 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2221 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2281 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     2341 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     2401 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     2461 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     2521 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     2581 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     2641 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     2701 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     2761 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     2821 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     2881 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     2941 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     3001 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     3061 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     3121 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     3181 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     3241 caaaatgccg caaaaaaggg aataagggcg acatggaaat gttgaatact catactcttc
     3301 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     3361 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     3421 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     3481 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     3541 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     3601 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
     3661 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     3721 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     3781 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     3841 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     3901 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     3961 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     4021 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
     4081 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     4141 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     4201 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta
     4261 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag
     4321 ctggctagcg tttaaactta agcttggtac cgagctcgga tccatggcct caaacgatta
     4381 tacccaacaa gcaacccaaa gctatggggc ctaccccacc cagcccgggc agggctattc
     4441 ccagcagagc agtcagccct acggacagca gagttacagt ggttatagcc agtccacgga
     4501 cacttcaggc tatggccaga gcagctattc ttcttatggc cagagccaga acacaggcta
     4561 tggaactcag tcaactcccc agggatatgg ctcgactggc ggctatggca gtagccagag
     4621 ctcccaatcg tcttacgggc agcagtcctc ctatcctggc tatggccagc agccagctcc
     4681 cagcagcacc tcgggaagtt acggtagcag ttctcagagc agcagctatg ggcagcccca
     4741 gagtgggagc tacagccagc agcctagcta tggtggacag cagcaaagct atggacagca
     4801 gcaaagctat aatccccctc agggctatgg acagcagaac cagtacaaca gcggtagcgg
     4861 cagcggtagc atggccacag acgcaatcga ccattactac cagaacagag acggcccccc
     4921 ttccaggagc tttagcgacg aatacgactc atacaataag acagcccctc tgccacctct
     4981 gagcgcctgc ccagagcgcg cccagacccc taacagcaag agaatcccca tcacacccaa
     5041 ttccgccaag agcacctccg agtctcctcg gagaatcaca ttctctcgcg acgcaggcag
     5101 ctccaccaag ggcagcaccc agacaagccc atccaaccag ggctccgtgt ctagccggaa
     5161 tggctctaac ggaaatggcc tgggcctgtc ctctgccggc aacttcgccg atttctttag
     5221 ctccgaggtg ttttatatcg tgctgcacaa tcccaccaca gcccacagat tcctgaagtt
     5281 ttgccagagc agggcctgtt ccgagaacct ggagtttctg cagaagatcg acgagtacaa
     5341 taggctgatc gatgaggcca cacgcctgct gtctaccatc tacagcacat acgtgagcag
     5401 cgaggcccct aagcagatca acatctctaa tagccacgcc aggcgcctga gccacgacat
     5461 caagggagca acccacaacg tgctgcccgg cctggaggat atcttcaatg gctcccagga
     5521 gcagatcgag aagctgctgg cctctgacct gtatcctaga tttgtgaagc accaggtgac
     5581 cgcatccgcc acaatggccc tggcaaacga caaggagagg tacccaggcc tgggcgattg
     5641 cttctgtctg acagacccac ggagagccga taatcccatc gtgttcgcaa gcgacggatt
     5701 cgtgagcgtg accggctatt cccgctctga tatcatccca aggaactgcc gcttcctgca
     5761 gggcagcttt accgaccggc aggccacaaa gaggctgcgc acctccatcg agaatcgcga
     5821 ggagacagtg gagctgctgc tgaactatcg gaagaatggc gatccctttt ggaacctgct
     5881 gtacgtgtcc cctctgctgg acggcaatgg cgatgtgaga ttctttctgg gcggccagat
     5941 caactgttct accacaatcc acagccggac cgacgtgctg agaatcctga gcctgaacga
     6001 cgaggataca ggctccttca tcgagggctc taataagacc cccagcgtgc ggtccaagga
     6061 gtctaacaat gcctccaact atggcaagac atccttcttt aagtctttta agaagtacaa
     6121 ggccaatacc gtggagatca gagacgaggc cggcatggag ggcgagctga tcgataggat
     6181 cggcaagctg cagttcaaga cacaggtgga ggagttttac accgcctatt ctaagtacct
     6241 ggtgctgagc tatcagccac agtcccagaa cctgatcgtg aagtacttca gccccggcat
     6301 cgtggatatg ctgaacctga atatgcctaa tggcctggtg gccccaatcg tggacaagga
     6361 tatcttcaag gtgctgacag agcactcccc ttcctctgtg ccacggacct ttaagaacgt
     6421 ggtgagggaa ggcatcaaag caggcagggc cgtgtctgtg gagacaggcc tgctgacagg
     6481 catcgaggag atcaagaagt ctcagaacat cttcagcggc accacagcca atcgggagag
     6541 aggctggaag aaggccgagg agaagtacgt gtgccacttt accccctgta aggatgagat
     6601 gggcagggtc ggctgggtgg tgctgactgt cgctcctaaa ctggaaagat ttgaattcgg
     6661 tggtggatca ggtggaggat ctatggtgag caagggcgag gaggataaca tggccatcat
     6721 caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga
     6781 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     6841 gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg
     6901 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc
     6961 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
     7021 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa
     7081 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga
     7141 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa
     7201 ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca
     7261 gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta
     7321 caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga
     7381 gctgtacaag taatgatcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     7441 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     7501 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     7561 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     7621 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     7681 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     7741 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     7801 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     7861 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     7921 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     7981 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     8041 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     8101 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     8161 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     8221 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     8281 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     8341 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     8401 gccgaggc
//
