LOCUS       sequence-200470-        6188 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses fusion of CRY2high mutant (CRY2PHR E490R) with mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6188)
  AUTHORS   Duan L, Hope J, Ong Q, Lou HY, Kim N, McCarthy C, Acero V, Lin MZ, 
            Cui B
  TITLE     Understanding CRY2 interactions for optical control of intracellular
            signaling.
  JOURNAL   Nat Commun. 2017 Sep 15;8(1):547. doi: 10.1038/s41467-017-00648-8.
  PUBMED    28916751
REFERENCE   2  (bases 1 to 6188)
  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: "Nat 
            Commun."; date: "2017-09-15"; volume: "8(1):547. doi"; pages: " 
            10.1038/s41467-017-00648-8"
COMMENT     Sequence Label: CRY2high-mCherry
FEATURES             Location/Qualifiers
     source          1..6188
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        59..362
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        363..566
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     517..537
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             623..2116
                     /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
                     SGVELGTNYAKPIVDIDTARELLAKAISRTRRAQIMIGAA"
     regulatory      2135..2144
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2141..2851
                     /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(2286..2304)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2554..2573
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2974..3095
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3011..3030)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3065..3084
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3102..3557)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3239..3260)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3451..3470
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3584..3688
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3690..4047
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3690..3881
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3713..3733)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3898..4033
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4082..4876
                     /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             4082..4876
                     /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(4136..4155)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4746..4765
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5064..5083)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5108..5155
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5484..6072
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5973..5992
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
       61 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      121 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      181 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      241 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      301 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      361 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      421 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      481 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      541 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
      601 ggactcagat ctcgaggcca ccatgaagat ggacaaaaag actatagttt ggtttagaag
      661 agacctaagg attgaggata atcctgcatt agcagcagct gctcacgaag gatctgtttt
      721 tcctgtcttc atttggtgtc ctgaagaaga aggacagttt tatcctggaa gagcttcaag
      781 atggtggatg aaacaatcac ttgctcactt atctcaatcc ttgaaggctc ttggatctga
      841 cctcacttta atcaaaaccc acaacacgat ttcagcgatc ttggattgta tccgcgttac
      901 cggtgctaca aaagtcgtct ttaaccacct ctatgatcct gtttcgttag ttcgggacca
      961 taccgtaaag gagaagctgg tggaacgtgg gatctctgtg caaagctaca atggagatct
     1021 attgtatgaa ccgtgggaga tatactgcga aaagggcaaa ccttttacga gtttcaattc
     1081 ttactggaag aaatgcttag atatgtcgat tgaatccgtt atgcttcctc ctccttggcg
     1141 gttgatgcca ataactgcag cggctgaagc gatttgggcg tgttcgattg aagaactagg
     1201 gctggagaat gaggccgaga aaccgagcaa tgcgttgtta actagagctt ggtctccagg
     1261 atggagcaat gctgataagt tactaaatga gttcatcgag aagcagttga tagattatgc
     1321 aaagaacagc aagaaagttg ttgggaattc tacttcacta ctttctccgt atctccattt
     1381 cggggaaata agcgtcagac acgttttcca gtgtgcccgg atgaaacaaa ttatatgggc
     1441 aagagataag aacagtgaag gagaagaaag tgcagatctt tttcttaggg gaatcggttt
     1501 aagagagtat tctcggtata tatgtttcaa cttcccgttt actcacgagc aatcgttgtt
     1561 gagtcatctt cggtttttcc cttgggatgc tgatgttgat aagttcaagg cctggagaca
     1621 aggcaggacc ggttatccgt tggtggatgc cggaatgaga gagctttggg ctaccggatg
     1681 gatgcataac agaataagag tgattgtttc aagctttgct gtgaagtttc ttctccttcc
     1741 atggaaatgg ggaatgaagt atttctggga tacacttttg gatgctgatt tggaatgtga
     1801 catccttggc tggcagtata tctctgggag tatccccgat ggccacgagc ttgatcgctt
     1861 ggacaatccc gcgttacaag gcgccaaata tgacccagaa ggtgagtaca taaggcaatg
     1921 gcttcccgag cttgcgagat tgccaactga atggatccat catccatggg acgctccttt
     1981 aaccgtactc aaagcttctg gtgtggaact cggaacaaac tatgcgaaac ccattgtaga
     2041 catcgacaca gctcgtgagc tactagctaa agctatttca agaacccgta gagcacagat
     2101 catgatcgga gcagcagccc gggatccacc ggtcgccacc atggtgagca agggcgagga
     2161 ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt
     2221 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
     2281 gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc
     2341 ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta
     2401 cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
     2461 cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt
     2521 gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg
     2581 ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa
     2641 gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa
     2701 ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac
     2761 ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc
     2821 caccggcggc atggacgagc tgtacaagta aagcggccgc gactctagat cataatcagc
     2881 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     2941 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     3001 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     3061 agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat
     3121 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     3181 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     3241 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     3301 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     3361 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     3421 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     3481 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     3541 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3601 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3661 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     3721 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3781 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3841 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3901 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3961 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     4021 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     4081 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4141 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4201 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4261 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4321 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     4381 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4441 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4501 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4561 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     4621 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4681 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4741 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4801 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4861 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     4921 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4981 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     5041 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     5101 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     5161 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     5221 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     5281 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     5341 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5401 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5461 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5521 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5581 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5641 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5701 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5761 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5821 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5881 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5941 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6001 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6061 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6121 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     6181 atgcatta
//
