LOCUS       sequence-224407-        7901 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes mCherry-tagged BcLOV4 for expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7901)
  AUTHORS   Glantz ST, Berlew EE, Jaber Z, Schuster BS, Gardner KH, Chow BY
  TITLE     Directly light-regulated binding of RGS-LOV photoreceptors to 
            anionic membrane phospholipids.
  JOURNAL   Proc Natl Acad Sci U S A. 2018 Aug 14;115(33):E7720-E7727. doi: 
            10.1073/pnas.1802832115. Epub 2018 Jul 31.
  PUBMED    30065115
REFERENCE   2  (bases 1 to 7901)
  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: 
            "10.1073/pnas.1802832115"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2018-08-14- 14"; volume: "115"; issue: "33"; pages: 
            "E7720-E7727"
COMMENT     Sequence Label: BcLOV4-mammalian-recoded_mCherry_pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..7901
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(19..25,26..43,44..49))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    64..85
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(202..219)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(373..961)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(453..472)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1132..1923,1924..1992))
                     /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     1755..1774
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1993..2097)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2172..2191)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2363..2742
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2743..2946
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2897..2917
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2991..3010
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2991..3009
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4875..5585
                     /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(5020..5038)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5288..5307
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(5620..5637)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5626..5850
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5896..6324
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5983..6002)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6193..6214
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6333..6353)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6338..6667
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6518..6653
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6580..6599
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6734..7528
                     /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(6788..6807)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7398..7417
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7702..7823
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7739..7758)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7793..7812
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7872..7888)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7872..7888)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(join(7885..7901,1..6))
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    join(7896..7901,1..11)
                     /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)."
ORIGIN
        1 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
       61 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
      121 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
      181 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
      241 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
      301 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
      361 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
      421 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
      481 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
      541 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
      601 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
      661 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
      721 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
      781 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
      841 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
      901 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
      961 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     1021 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     1081 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     1141 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     1201 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     1261 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     1321 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     1381 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     1441 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     1501 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     1561 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     1621 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     1681 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     1741 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     1801 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     1861 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     1921 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga catggaaatg
     1981 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     2041 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     2101 atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg
     2161 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct
     2221 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
     2281 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
     2341 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
     2401 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     2461 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     2521 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     2581 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     2641 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     2701 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     2761 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     2821 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     2881 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     2941 agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact
     3001 cactataggg agacccaagc tggctagcgt ttaaacttaa gcttggtacc gagctcggat
     3061 ccatggccac agacgcaatc gaccattact accagaacag agacggcccc ccttccagga
     3121 gctttagcga cgaatacgac tcatacaata agacagcccc tctgccacct ctgagcgcct
     3181 gcccagagcg cgcccagacc cctaacagca agagaatccc catcacaccc aattccgcca
     3241 agagcacctc cgagtctcct cggagaatca cattctctcg cgacgcaggc agctccacca
     3301 agggcagcac ccagacaagc ccatccaacc agggctccgt gtctagccgg aatggctcta
     3361 acggaaatgg cctgggcctg tcctctgccg gcaacttcgc cgatttcttt agctccgagg
     3421 tgttttatat cgtgctgcac aatcccacca cagcccacag attcctgaag ttttgccaga
     3481 gcagggcctg ttccgagaac ctggagtttc tgcagaagat cgacgagtac aataggctga
     3541 tcgatgaggc cacacgcctg ctgtctacca tctacagcac atacgtgagc agcgaggccc
     3601 ctaagcagat caacatctct aatagccacg ccaggcgcct gagccacgac atcaagggag
     3661 caacccacaa cgtgctgccc ggcctggagg atatcttcaa tggctcccag gagcagatcg
     3721 agaagctgct ggcctctgac ctgtatccta gatttgtgaa gcaccaggtg accgcatccg
     3781 ccacaatggc cctggcaaac gacaaggaga ggtacccagg cctgggcgat tgcttctgtc
     3841 tgacagaccc acggagagcc gataatccca tcgtgttcgc aagcgacgga ttcgtgagcg
     3901 tgaccggcta ttcccgctct gatatcatcc caaggaactg ccgcttcctg cagggcagct
     3961 ttaccgaccg gcaggccaca aagaggctgc gcacctccat cgagaatcgc gaggagacag
     4021 tggagctgct gctgaactat cggaagaatg gcgatccctt ttggaacctg ctgtacgtgt
     4081 cccctctgct ggacggcaat ggcgatgtga gattctttct gggcggccag atcaactgtt
     4141 ctaccacaat ccacagccgg accgacgtgc tgagaatcct gagcctgaac gacgaggata
     4201 caggctcctt catcgagggc tctaataaga cccccagcgt gcggtccaag gagtctaaca
     4261 atgcctccaa ctatggcaag acatccttct ttaagtcttt taagaagtac aaggccaata
     4321 ccgtggagat cagagacgag gccggcatgg agggcgagct gatcgatagg atcggcaagc
     4381 tgcagttcaa gacacaggtg gaggagtttt acaccgccta ttctaagtac ctggtgctga
     4441 gctatcagcc acagtcccag aacctgatcg tgaagtactt cagccccggc atcgtggata
     4501 tgctgaacct gaatatgcct aatggcctgg tggccccaat cgtggacaag gatatcttca
     4561 aggtgctgac agagcactcc ccttcctctg tgccacggac ctttaagaac gtggtgaggg
     4621 aaggcatcaa agcaggcagg gccgtgtctg tggagacagg cctgctgaca ggcatcgagg
     4681 agatcaagaa gtctcagaac atcttcagcg gcaccacagc caatcgggag agaggctgga
     4741 agaaggccga ggagaagtac gtgtgccact ttaccccctg taaggatgag atgggcaggg
     4801 tcggctgggt ggtgctgact gtcgctccta aactggaaag atttgaattc ggtggtggat
     4861 caggtggagg atctatggtg agcaagggcg aggaggataa catggccatc atcaaggagt
     4921 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
     4981 gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg
     5041 gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac ggctccaagg
     5101 cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct
     5161 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
     5221 cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct
     5281 ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc gagcggatgt
     5341 accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg aaggacggcg
     5401 gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg cagctgcccg
     5461 gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac tacaccatcg
     5521 tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac gagctgtaca
     5581 agtaatgatc tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     5641 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     5701 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     5761 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     5821 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     5881 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     5941 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     6001 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     6061 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     6121 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     6181 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     6241 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     6301 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     6361 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     6421 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     6481 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     6541 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     6601 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     6661 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
     6721 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     6781 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     6841 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     6901 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     6961 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     7021 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     7081 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     7141 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     7201 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     7261 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     7321 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     7381 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     7441 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     7501 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     7561 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     7621 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     7681 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     7741 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     7801 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     7861 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt a
//
