LOCUS       sequence-204322-        7291 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cry2-mCherry-PKA W196R/Y204A in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7291)
  AUTHORS   O'Banion CP, Priestman MA, Hughes RM, Herring LE, Capuzzi SJ, 
            Lawrence DS
  TITLE     Design and Profiling of a Subcellular Targeted Optogenetic 
            cAMP-Dependent Protein Kinase.
  JOURNAL   Cell Chem Biol. 2017 Oct 25. pii: S2451-9456(17)30355-0. doi: 
            10.1016/j.chembiol.2017.09.011.
  PUBMED    29104065
REFERENCE   2  (bases 1 to 7291)
  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: "Cell Chem 
            Biol. 2017 Oct 25. pii: S2451-9456(17)30355-0. doi: 
            10.1016/j.chembiol.2017.09.011."
COMMENT     Sequence Label: optoPKAW196R/Y204A
FEATURES             Location/Qualifiers
     source          1..7291
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             625..2118
                     /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      2137..2146
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2143..2850
                     /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(2288..2306)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2556..2575
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4079..4200
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4116..4135)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4170..4189
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4207..4662)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4344..4365)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4556..4575
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4689..4793
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4795..5152
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4795..4986
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4818..4838)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5003..5138
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5187..5981
                     /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             5187..5981
                     /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(5241..5260)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5851..5870
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6169..6188)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6213..6260
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6589..7177
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7078..7097
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgaggc caccatgaag atggacaaaa agactatagt ttggtttaga
      661 agagacctaa ggattgagga taatcctgca ttagcagcag ctgctcacga aggatctgtt
      721 tttcctgtct tcatttggtg tcctgaagaa gaaggacagt tttatcctgg aagagcttca
      781 agatggtgga tgaaacaatc acttgctcac ttatctcaat ccttgaaggc tcttggatct
      841 gacctcactt taatcaaaac ccacaacacg atttcagcga tcttggattg tatccgcgtt
      901 accggtgcta caaaagtcgt ctttaaccac ctctatgatc ctgtttcgtt agttcgggac
      961 cataccgtaa aggagaagct ggtggaacgt gggatctctg tgcaaagcta caatggagat
     1021 ctattgtatg aaccgtggga gatatactgc gaaaagggca aaccttttac gagtttcaat
     1081 tcttactgga agaaatgctt agatatgtcg attgaatccg ttatgcttcc tcctccttgg
     1141 cggttgatgc caataactgc agcggctgaa gcgatttggg cgtgttcgat tgaagaacta
     1201 gggctggaga atgaggccga gaaaccgagc aatgcgttgt taactagagc ttggtctcca
     1261 ggatggagca atgctgataa gttactaaat gagttcatcg agaagcagtt gatagattat
     1321 gcaaagaaca gcaagaaagt tgttgggaat tctacttcac tactttctcc gtatctccat
     1381 ttcggggaaa taagcgtcag acacgttttc cagtgtgccc ggatgaaaca aattatatgg
     1441 gcaagagata agaacagtga aggagaagaa agtgcagatc tttttcttag gggaatcggt
     1501 ttaagagagt attctcggta tatatgtttc aacttcccgt ttactcacga gcaatcgttg
     1561 ttgagtcatc ttcggttttt cccttgggat gctgatgttg ataagttcaa ggcctggaga
     1621 caaggcagga ccggttatcc gttggtggat gccggaatga gagagctttg ggctaccgga
     1681 tggatgcata acagaataag agtgattgtt tcaagctttg ctgtgaagtt tcttctcctt
     1741 ccatggaaat ggggaatgaa gtatttctgg gatacacttt tggatgctga tttggaatgt
     1801 gacatccttg gctggcagta tatctctggg agtatccccg atggccacga gcttgatcgc
     1861 ttggacaatc ccgcgttaca aggcgccaaa tatgacccag aaggtgagta cataaggcaa
     1921 tggcttcccg agcttgcgag attgccaact gaatggatcc atcatccatg ggacgctcct
     1981 ttaaccgtac tcaaagcttc tggtgtggaa ctcggaacaa actatgcgaa acccattgta
     2041 gacatcgaca cagctcgtga gctactagct aaagctattt caagaacccg tgaagcacag
     2101 atcatgatcg gagcagcagc ccgggatcca ccggtcgcca ccatggtgag caagggcgag
     2161 gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc
     2221 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc
     2281 cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg
     2341 tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac
     2401 tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac
     2461 ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag
     2521 gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg
     2581 ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc
     2641 aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac
     2701 aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc
     2761 acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac
     2821 tccaccggcg gcatggacga gctgtacaag agtgctggtg gtagtgctgg tggtagtgct
     2881 ggtggtagtg ctggtggtat gggcaacgcc gccgccgcca agaagggcag cgagcaggag
     2941 agcgtgaaag agttcctagc caaagccaag gaagatttcc tgaaaaaatg ggagacccct
     3001 tctcagaata cagcccagtt ggatcagttt gatagaatca agacccttgg caccggctcc
     3061 tttgggcgag tgatgctggt gaagcacaag gagagtggga accactacgc catgaagatc
     3121 ttagacaagc agaaggtggt gaagctaaag cagatcgagc acactctgaa tgagaagcgc
     3181 atcctgcagg ccgtcaactt cccgttcctg gtcaaacttg aattctcctt caaggacaac
     3241 tcaaacctgt atatggtcat ggagtatgta gctggtggcg agatgttctc ccacctacgg
     3301 cggattggaa ggttcagcga gccccatgcc cgtttctacg cggcgcagat cgtcctgacc
     3361 tttgagtatc tgcactccct ggacctcatc taccgggacc tgaagcccga gaatcttctc
     3421 atcgaccagc agggctatat tcaggtgaca gacttcggtt ttgccaagcg tgtgaaaggc
     3481 cgtactcgga ccttgtgtgg gacccctgag gccttggccc ccgagattat cctgagcaaa
     3541 ggctacaaca aggctgtgga ctggtgggct ctcggagtcc tcatctacga gatggctgct
     3601 ggttacccac ccttcttcgc tgaccagcct atccagatct atgagaaaat cgtctctggg
     3661 aaggtgcggt tcccatccca cttcagctct gacttgaagg acctgctgcg gaaccttctg
     3721 caagtggatc tcaccaagcg ctttgggaac ctcaagaacg gggtcaatga catcaagaac
     3781 cacaagtggt ttgccacgac tgactggatt gccatctatc agagaaaggt ggaagctccc
     3841 ttcataccaa agtttaaagg ccctggggac acgagtaact ttgacgacta tgaggaggaa
     3901 gagatccggg tctccatcaa tgagaagtgt ggcaaggagt ttactgagtt ttaatgagcg
     3961 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     4021 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     4081 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4141 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4201 aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     4261 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     4321 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     4381 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     4441 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     4501 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     4561 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     4621 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     4681 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4741 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     4801 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4861 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4921 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4981 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     5041 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     5101 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     5161 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     5221 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     5281 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     5341 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     5401 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     5461 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     5521 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     5581 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     5641 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     5701 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     5761 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     5821 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     5881 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     5941 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     6001 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     6061 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     6121 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     6181 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     6241 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     6301 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     6361 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     6421 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6481 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6541 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6601 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6661 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6721 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6781 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6841 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6901 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6961 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     7021 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     7081 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     7141 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     7201 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     7261 ttctgtggat aaccgtatta ccgccatgca t
//
