LOCUS       sequence-216307-        7898 bp    DNA     circular SYN 14-APR-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 7898)
  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 7898)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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..7898
                     /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..958)
                     /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(1129..1920,1921..1989))
                     /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     1752..1771
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1990..2094)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(2169..2188)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2360..2739
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2740..2943
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2894..2914
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2988..3007
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2988..3006
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4872..5582
                     /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(5017..5035)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5285..5304
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(5617..5634)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5623..5847
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5893..6321
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5980..5999)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6190..6211
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6330..6350)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6335..6664
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6515..6650
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6577..6596
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6731..7525
                     /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(6785..6804)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7395..7414
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7699..7820
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7736..7755)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7790..7809
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7869..7885)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7869..7885)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(join(7882..7898,1..6))
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    join(7893..7898,1..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 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
      961 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     1021 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     1081 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     1141 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     1201 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     1261 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     1321 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     1381 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     1441 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     1501 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     1561 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     1621 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     1681 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     1741 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     1801 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     1861 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     1921 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     1981 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     2041 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     2101 tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggtgc
     2161 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     2221 gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg
     2281 accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta
     2341 cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg
     2401 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     2461 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     2521 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     2581 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     2641 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     2701 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     2761 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     2821 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     2881 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     2941 gctctctggc taactagaga acccactgct tactggctta tcgaaattaa tacgactcac
     3001 tatagggaga cccaagctgg ctagcgttta aacttaagct tggtaccgag ctcggatcca
     3061 tggccacaga cgcaatcgac cattactacc agaacagaga cggcccccct tccaggagct
     3121 ttagcgacga atacgactca tacaataaga cagcccctct gccacctctg agcgcctgcc
     3181 cagagcgcgc ccagacccct aacagcaaga gaatccccat cacacccaat tccgccaaga
     3241 gcacctccga gtctcctcgg agaatcacat tctctcgcga cgcaggcagc tccaccaagg
     3301 gcagcaccca gacaagccca tccaaccagg gctccgtgtc tagccggaat ggctctaacg
     3361 gaaatggcct gggcctgtcc tctgccggca acttcgccga tttctttagc tccgaggtgt
     3421 tttatatcgt gctgcacaat cccaccacag cccacagatt cctgaagttt tgccagagca
     3481 gggcctgttc cgagaacctg gagtttctgc agaagatcga cgagtacaat aggctgatcg
     3541 atgaggccac acgcctgctg tctaccatct acagcacata cgtgagcagc gaggccccta
     3601 agcagatcaa catctctaat agccacgcca ggcgcctgag ccacgacatc aagggagcaa
     3661 cccacaacgt gctgcccggc ctggaggata tcttcaatgg ctcccaggag cagatcgaga
     3721 agctgctggc ctctgacctg tatcctagat ttgtgaagca ccaggtgacc gcatccgcca
     3781 caatggccct ggcaaacgac aaggagaggt acccaggcct gggcgattgc ttctgtctga
     3841 cagacccacg gagagccgat aatcccatcg tgttcgcaag cgacggattc gtgagcgtga
     3901 ccggctattc ccgctctgat atcatcccaa ggaactgccg cttcctgcag ggcagcttta
     3961 ccgaccggca ggccacaaag aggctgcgca cctccatcga gaatcgcgag gagacagtgg
     4021 agctgctgct gaactatcgg aagaatggcg atcccttttg gaacctgctg tacgtgtccc
     4081 ctctgctgga cggcaatggc gatgtgagat tctttctggg cggccagatc aactgttcta
     4141 ccacaatcca cagccggacc gacgtgctga gaatcctgag cctgaacgac gaggatacag
     4201 gctccttcat cgagggctct aataagaccc ccagcgtgcg gtccaaggag tctaacaatg
     4261 cctccaacta tggcaagaca tccttcttta agtcttttaa gaagtacaag gccaataccg
     4321 tggagatcag agacgaggcc ggcatggagg gcgagctgat cgataggatc ggcaagctgc
     4381 agttcaagac acaggtggag gagttttaca ccgcctattc taagtacctg gtgctgagct
     4441 atcagccaca gtcccagaac ctgatcgtga agtacttcag ccccggcatc gtggatatgc
     4501 tgaacctgaa tatgcctaat ggcctggtgg ccccaatcgt ggacaaggat atcttcaagg
     4561 tgctgacaga gcactcccct tcctctgtgc cacggacctt taagaacgtg gtgagggaag
     4621 gcatcaaagc aggcagggcc gtgtctgtgg agacaggcct gctgacaggc atcgaggaga
     4681 tcaagaagtc tcagaacatc ttcagcggca ccacagccaa tcgggagaga ggctggaaga
     4741 aggccgagga gaagtacgtg tgccacttta ccccctgtaa ggatgagatg ggcagggtcg
     4801 gctgggtggt gctgactgtc gctcctaaac tggaaagatt tgaattcggt ggtggatcag
     4861 gtggaggatc tatggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca
     4921 tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg
     4981 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg
     5041 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct
     5101 acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca
     5161 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct
     5221 ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg
     5281 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc
     5341 ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc
     5401 actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg
     5461 cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg
     5521 aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt
     5581 aatgatctag agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     5641 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     5701 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     5761 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     5821 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     5881 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     5941 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     6001 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     6061 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     6121 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     6181 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     6241 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     6301 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     6361 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     6421 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     6481 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     6541 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     6601 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     6661 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     6721 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     6781 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     6841 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     6901 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     6961 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     7021 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     7081 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     7141 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     7201 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     7261 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     7321 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     7381 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     7441 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     7501 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     7561 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     7621 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     7681 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     7741 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     7801 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     7861 taatcatggt catagctgtt tcctgtgtga aattgtta
//
