LOCUS       sequence-382847-        7336 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses a fluorescently-tagged, optogenetically activated 
            PKC-epsilon. Used in tandem with CIBN-CAAX.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7336)
  AUTHORS   Gada KD, Kawano T, Plant LD, Logothetis DE
  TITLE     An optogenetic tool to recruit individual PKC isozymes to the cell 
            surface and promote specific phosphorylation of membrane proteins.
  JOURNAL   J Biol Chem. 2022 May;298(5):101893. doi: 10.1016/j.jbc.2022.101893.
            Epub 2022 Mar 31.
  PUBMED    35367414
REFERENCE   2  (bases 1 to 7336)
  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: "J Biol 
            Chem."; date: "2022-05"; volume: "298(5):101893. doi"; pages: " 
            10.1016/j.jbc.2022.101893. Epub 2022 Mar 31"
COMMENT     Sequence Label: mCherry-CRY2–mPKC-epsilon-CAT-HA
FEATURES             Location/Qualifiers
     source          1..7336
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        63..366
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        367..570
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     521..541
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      608..617
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             614..1321
                     /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(759..777)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1027..1046
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1367..2860
                     /codon_start=1
                     /product="N-terminal photolyase homology region of 
                     Arabidopsis thaliana cryptochrome 2"
                     /label=CRY2PHR
                     /note="binds to CIB1 upon exposure to blue light (Kennedy 
                     et al., 2010)"
                     /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
                     FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
                     DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
                     ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
                     EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
                     ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAA"
     regulatory      2909..2918
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3959..3985
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    4126..4247
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4163..4182)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4217..4236
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4254..4709)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4391..4412)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4603..4622
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4736..4840
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4842..5199
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4842..5033
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4865..4885)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5050..5185
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5234..6028
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5234..6028
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5288..5307)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5898..5917
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6216..6235)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6260..6307
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6636..7224
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7125..7144
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
       61 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      121 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      181 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      241 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      301 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      361 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      421 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      481 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      541 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc
      601 taccggtgcc accatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt
      661 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
      721 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
      781 tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc
      841 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
      901 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
      961 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
     1021 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta
     1081 ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg
     1141 ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
     1201 cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt
     1261 ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     1321 gagatctcga agcgcggccg cgggagcagg aggagcagct cgagctatga agatggacaa
     1381 aaagactata gtttggttta gaagagacct aaggattgag gataatcctg cattagcagc
     1441 agctgctcac gaaggatctg tttttcctgt cttcatttgg tgtcctgaag aagaaggaca
     1501 gttttatcct ggaagagctt caagatggtg gatgaaacaa tcacttgctc acttatctca
     1561 atccttgaag gctcttggat ctgacctcac tttaatcaaa acccacaaca cgatttcagc
     1621 gatcttggat tgtatccgcg ttaccggtgc tacaaaagtc gtctttaacc acctctatga
     1681 tcctgtttcg ttagttcggg accataccgt aaaggagaag ctggtggaac gtgggatctc
     1741 tgtgcaaagc tacaatggag atctattgta tgaaccgtgg gagatatact gcgaaaaggg
     1801 caaacctttt acgagtttca attcttactg gaagaaatgc ttagatatgt cgattgaatc
     1861 cgttatgctt cctcctcctt ggcggttgat gccaataact gcagcggctg aagcgatttg
     1921 ggcgtgttcg attgaagaac tagggctgga gaatgaggcc gagaaaccga gcaatgcgtt
     1981 gttaactaga gcttggtctc caggatggag caatgctgat aagttactaa atgagttcat
     2041 cgagaagcag ttgatagatt atgcaaagaa cagcaagaaa gttgttggga attctacttc
     2101 actactttct ccgtatctcc atttcgggga aataagcgtc agacacgttt tccagtgtgc
     2161 ccggatgaaa caaattatat gggcaagaga taagaacagt gaaggagaag aaagtgcaga
     2221 tctttttctt aggggaatcg gtttaagaga gtattctcgg tatatatgtt tcaacttccc
     2281 gtttactcac gagcaatcgt tgttgagtca tcttcggttt ttcccttggg atgctgatgt
     2341 tgataagttc aaggcctgga gacaaggcag gaccggttat ccgttggtgg atgccggaat
     2401 gagagagctt tgggctaccg gatggatgca taacagaata agagtgattg tttcaagctt
     2461 tgctgtgaag tttcttctcc ttccatggaa atggggaatg aagtatttct gggatacact
     2521 tttggatgct gatttggaat gtgacatcct tggctggcag tatatctctg ggagtatccc
     2581 cgatggccac gagcttgatc gcttggacaa tcccgcgtta caaggcgcca aatatgaccc
     2641 agaaggtgag tacataaggc aatggcttcc cgagcttgcg agattgccaa ctgaatggat
     2701 ccatcatcca tgggacgctc ctttaaccgt actcaaagct tctggtgtgg aactcggaac
     2761 aaactatgcg aaacccattg tagacatcga cacagctcgt gagctactag ctaaagctat
     2821 ttcaagaacc cgtgaagcac agatcatgat cggagcagca gctcgaggcg cagcagcggg
     2881 agcaggagga gcaggtcgga tcctcgaggc caccatggaa ttggtccggc caggccaggc
     2941 caagcgcttg gggctggatg agttcaactt catcaaagtg ttgggcaaag gcagctttgg
     3001 caaggtcatg ttggcggaac tcaaaggcaa agatgaagtc tacgctgtga aggtcttgaa
     3061 gaaggacgtt atcctacaag acgatgatgt ggactgcaca atgacagaga agaggatttt
     3121 ggctctggct cggaaacacc cttatctaac ccaactctat tgctgcttcc agaccaagga
     3181 ccgcctcttc ttcgtcatgg aatatgtaaa tggtggagac ctcatgttcc agattcagcg
     3241 gtcccgaaaa tttgatgagc ctcgttctcg gttctatgcc gcagaggtca catcggccct
     3301 catgtttctc caccagcatg gagtgatcta cagggatttg aaactggaca acatccttct
     3361 agatgcagaa ggccactgca agctggctga ctttgggatg tgcaaggaag ggattatgaa
     3421 tggtgtgaca actaccacct tctgtgggac tcctgactac atagctccag agatcctaca
     3481 ggagttggag tacggcccct cagtggactg gtgggccctg ggggtgctga tgtacgagat
     3541 gatggctggg cagcccccct ttgaagctga caacgaggac gacttgttcg aatccatcct
     3601 tcatgatgat gttctctatc ctgtctggct tagcaaggaa gctgtcagca tcctgaaagc
     3661 tttcatgacc aagaacccgc acaagcgcct gggctgtgtg gcagcgcaga acggggagga
     3721 cgccatcaag caacatccat tcttcaagga gattgactgg gtactgctgg agcagaagaa
     3781 aatcaagccc cccttcaagc cgagaattaa aaccaaaaga gatgtcaata actttgacca
     3841 agactttacg cgggaagagc caatacttac acttgtggat gaagcaatca ttaagcagat
     3901 caaccaggaa gaattcaaag gcttctccta ctttggtgaa gacctgatgc cccaattcta
     3961 tccttacgac gtgcctgact acgcctaagg atcccgggat ccaccggatc tagataactg
     4021 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     4081 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     4141 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4201 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt
     4261 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     4321 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     4381 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     4441 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     4501 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     4561 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     4621 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     4681 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     4741 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     4801 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     4861 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4921 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4981 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     5041 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     5101 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     5161 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     5221 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     5281 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     5341 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     5401 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     5461 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     5521 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     5581 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     5641 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     5701 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     5761 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     5821 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     5881 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     5941 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     6001 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     6061 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     6121 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     6181 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     6241 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     6301 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     6361 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     6421 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     6481 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6541 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6601 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6661 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6721 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6781 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6841 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6901 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6961 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     7021 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     7081 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     7141 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     7201 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     7261 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     7321 cgtattaccg ccatgc
//
