LOCUS       sequence-90204-e        6243 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Light insensitive CRY2 mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6243)
  AUTHORS   Lee S, Park H, Kyung T, Kim NY, Kim S, Kim J, Heo WD
  TITLE     Reversible protein inactivation by optogenetic trapping in cells.
  JOURNAL   Nat Methods. 2014 Jun;11(6):633-6. doi: 10.1038/nmeth.2940. Epub 
            2014 May 4.
  PUBMED    24793453
REFERENCE   2  (bases 1 to 6243)
  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: 
            "10.1038/nmeth.2940"; journalName: "Nat Methods"; date: "2014-06"; 
            volume: "11"; issue: "6"; pages: "633-6"
COMMENT     Sequence Label: pCMV-CRY2(D387A)-mCherry
FEATURES             Location/Qualifiers
     source          1..6243
                     /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"
     regulatory      2128..2137
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2134..2841
                     /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(2279..2297)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2547..2566
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    2842..2907
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    3031..3152
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3068..3087)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3122..3141
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3159..3614)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3296..3317)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3508..3527
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3641..3745
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3747..4104
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3747..3938
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3770..3790)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3955..4090
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4139..4933
                     /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             4139..4933
                     /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(4193..4212)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4803..4822
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5121..5140)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5165..5212
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5541..6129
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6030..6049
                     /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 ccggtcgcca ccatgaagat ggacaaaaag accatcgtct ggtttcggag agatttgaga
      661 atagaagata atcccgcgct cgccgccgcg gcccacgagg gttccgtctt ccccgttttc
      721 atttggtgtc ctgaagaaga aggccagttt tatcccggaa gggcctctag gtggtggatg
      781 aagcaaagtc tggcccatct tagccagtca ctgaaagcac tgggcagtga tcttaccctg
      841 atcaagacac acaataccat ctctgccatc ctcgactgca tcagggtgac cggcgcaacg
      901 aaagtcgtgt ttaaccacct gtacgatcca gttagtctgg tgcgcgacca cactgtgaag
      961 gagaagctgg tggaacgggg gatcagtgtg cagagctaca acggggacct tctgtacgag
     1021 ccatgggaga tctattgcga gaaagggaaa ccgttcacct ccttcaacag ttactggaag
     1081 aaatgtttgg atatgtcaat agagtccgtt atgttgcccc ctccctggag actgatgccg
     1141 attactgctg ctgcagaggc catctgggcc tgctccatcg aggaactcgg tctggaaaat
     1201 gaagcagaaa agccaagcaa tgcacttctc actagagcct ggagccccgg ctggtctaat
     1261 gccgacaagc tgcttaacga gttcatcgaa aaacaactga ttgactacgc gaagaactcc
     1321 aagaaagtgg taggtaactc aactagcttg ctctctccat atctccattt tggcgagatt
     1381 tctgtccgcc atgtatttca gtgcgctcgg atgaaacaga ttatctgggc tcgcgataaa
     1441 aacagcgaag gcgaagaaag cgccgatctg ttcctgcgag ggatcggact tcgggaatac
     1501 tcccggtata tatgtttcaa ctttccattc acacacgagc agagtctgtt gtcccacctc
     1561 aggttcttcc cctgggacgc cgatgtcgac aaattcaagg catggagaca gggaaggaca
     1621 gggtacccac tcgtggatgc tggcatgaga gagctctggg ctacaggctg gatgcacaac
     1681 cgcatccggg taatcgtgtc ctcatttgct gtcaagtttc tgctcctgcc ttggaaatgg
     1741 ggaatgaagt acttttggga tacccttctc gccgccgact tggagtgtga cattctggga
     1801 tggcaatata ttagcgggtc aattcctgac ggccatgagt tggacaggtt ggacaatccg
     1861 gccttgcagg gagctaagta tgatcccgaa ggagagtata ttcgacagtg gctccccgag
     1921 ctggcccgac ttcctacgga gtggattcac catccttggg acgcaccact gacagtgctc
     1981 aaggcaagcg gggtggagct gggcaccaat tacgctaagc ctatagttga tatagataca
     2041 gcacgcgagc tgctggctaa agcgatctct cgcactcggg aggcgcagat tatgatcggt
     2101 gctgccggca gcggcagccc accggtcgcc accatggtga gcaagggcga ggaggataac
     2161 atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc
     2221 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     2281 aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag
     2341 ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag
     2401 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg
     2461 gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg
     2521 cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     2581 gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg
     2641 ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag
     2701 aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac
     2761 aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc
     2821 ggcatggacg agctgtacaa gtccggactc agatctcgag ctcaagcttc gaattctgca
     2881 gtcgacggta ccgcgggccc gggatccacc ggatctagat aactgatcat aatcagccat
     2941 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     3001 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     3061 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3121 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt
     3181 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     3241 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     3301 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     3361 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     3421 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     3481 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     3541 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     3601 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3661 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3721 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     3781 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3841 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3901 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3961 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4021 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     4081 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     4141 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     4201 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     4261 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     4321 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     4381 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     4441 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     4501 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     4561 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     4621 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     4681 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4741 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4801 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4861 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4921 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     4981 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     5041 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     5101 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     5161 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     5221 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     5281 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     5341 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     5401 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5461 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5521 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5581 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5641 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     5701 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5761 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5821 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5881 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     5941 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6001 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6061 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6121 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6181 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     6241 cat
//
