LOCUS       sequence-183928-        7373 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CRY2 (full-length, with endogenous NLS) fused to Gal4 
            (aa1-65 of Gal4BD), downstream of mCherry-IRES.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7373)
  AUTHORS   Pathak GP, Spiltoir JI, Hoglund C, Polstein LR, Heine-Koskinen S, 
            Gersbach CA, Rossi J, Tucker CL
  TITLE     Bidirectional approaches for optogenetic regulation of gene 
            expression in mammalian cells using Arabidopsis cryptochrome 2.
  JOURNAL   Nucleic Acids Res. 2017 Apr 20. doi: 10.1093/nar/gkx260.
  PUBMED    28431041
REFERENCE   2  (bases 1 to 7373)
  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: "Nucleic 
            Acids Res. 2017 Apr 20. doi: 10.1093/nar/gkx260."
COMMENT     Sequence Label: CRY-Gal∆DD (B1013)
FEATURES             Location/Qualifiers
     source          1..7373
                     /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      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             613..1320
                     /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(758..776)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1026..1045
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(1922..1938)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1923..1939)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             1950..3785
                     /codon_start=1
                     /product="Arabidopsis thaliana cryptochrome 2"
                     /label=CRY2
                     /note="binds to CIB1 upon exposure to blue light (Kennedy 
                     et al., 2010)"
                     /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
                     FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
                     DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
                     ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
                     EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
                     ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAAPDEIVADSFEALGANTIKE
                     PGLCPSVSSNDQQVPSAVRYNGSKRVKPEEEEERDMKKSRGFDERELFSTAESSSSSSV
                     FFVSQSCSLASEGKNLEGIQDSSDQITTSLGKNGCK"
     polyA_signal    4161..4282
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4198..4217)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4252..4271
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4289..4744)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4426..4447)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4638..4657
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4771..4875
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4877..5234
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4877..5068
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4900..4920)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5085..5220
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5269..6063
                     /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             5269..6063
                     /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(5323..5342)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5933..5952
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6251..6270)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6295..6342
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6671..7259
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7160..7179
                     /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 ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
      661 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
      721 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
      781 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
      841 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
      901 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
      961 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     1021 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     1081 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     1141 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     1201 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     1261 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     1321 tgtggcggct agtactccgg tattgcggta cccttgtacg cctgttttat actcccttcc
     1381 cgtaacttag acgcacaaaa ccaagttcaa tagaaggggg tacaaaccag taccaccacg
     1441 aacaagcact tctgtttccc cggtgatgtc gtatagactg cttgcgtggt tgaaagcgac
     1501 ggatccgtta tccgcttatg tacttcgaga agcccagtac cacctcggaa tcttcgatgc
     1561 gttgcgctca gcactcaacc ccagagtgta gcttaggctg atgagtctgg acatccctca
     1621 ccggtgacgg tggtccaggc tgcgttggcg gcctacctat ggctaacgcc atgggacgct
     1681 agttgtgaac aaggtgtgaa gagcctattg agctacataa gaatcctccg gcccctgaat
     1741 gcggctaatc ccaacctcgg agcaggtggt cacaaaccag tgattggcct gtcgtaacgc
     1801 gcaagtccgt ggcggaaccg actactttgg gtgtccgtgt ttccttttat tttattgtgg
     1861 ctgcttatgg tgacaatcac agattgttat cataaagcga attggatagg atcaagctta
     1921 tcgataccgt cgacctcgag ctcgccacca tgaagatgga caaaaagact atagtttggt
     1981 ttagaagaga cctaaggatt gaggataatc ctgcattagc agcagctgct cacgaaggat
     2041 ctgtttttcc tgtcttcatt tggtgtcctg aagaagaagg acagttttat cctggaagag
     2101 cttcaagatg gtggatgaaa caatcacttg ctcacttatc tcaatccttg aaggctcttg
     2161 gatctgacct cactttaatc aaaacccaca acacgatttc agcgatcttg gattgtatcc
     2221 gcgttaccgg tgctacaaaa gtcgtcttta accacctcta tgatcctgtt tcgttagttc
     2281 gggaccatac cgtaaaggag aagctggtgg aacgtgggat ctctgtgcaa agctacaatg
     2341 gagatctatt gtatgaaccg tgggagatat actgcgaaaa gggcaaacct tttacgagtt
     2401 tcaattctta ctggaagaaa tgcttagata tgtcgattga atccgttatg cttcctcctc
     2461 cttggcggtt gatgccaata actgcagcgg ctgaagcgat ttgggcgtgt tcgattgaag
     2521 aactagggct ggagaatgag gccgagaaac cgagcaatgc gttgttaact agagcttggt
     2581 ctccaggatg gagcaatgct gataagttac taaatgagtt catcgagaag cagttgatag
     2641 attatgcaaa gaacagcaag aaagttgttg ggaattctac ttcactactt tctccgtatc
     2701 tccatttcgg ggaaataagc gtcagacacg ttttccagtg tgcccggatg aaacaaatta
     2761 tatgggcaag agataagaac agtgaaggag aagaaagtgc agatcttttt cttaggggaa
     2821 tcggtttaag agagtattct cggtatatat gtttcaactt cccgtttact cacgagcaat
     2881 cgttgttgag tcatcttcgg tttttccctt gggatgctga tgttgataag ttcaaggcct
     2941 ggagacaagg caggaccggt tatccgttgg tggatgccgg aatgagagag ctttgggcta
     3001 ccggatggat gcataacaga ataagagtga ttgtttcaag ctttgctgtg aagtttcttc
     3061 tccttccatg gaaatgggga atgaagtatt tctgggatac acttttggat gctgatttgg
     3121 aatgtgacat ccttggctgg cagtatatct ctgggagtat ccccgatggc cacgagcttg
     3181 atcgcttgga caatcccgcg ttacaaggcg ccaaatatga cccagaaggt gagtacataa
     3241 ggcaatggct tcccgagctt gcgagattgc caactgaatg gatccatcat ccatgggacg
     3301 ctcctttaac cgtactcaaa gcttctggtg tggaactcgg aacaaactat gcgaaaccca
     3361 ttgtagacat cgacacagct cgtgagctac tagctaaagc tatttcaaga acccgtgaag
     3421 cacagatcat gatcggagca gcacctgatg agattgtagc agatagcttc gaggccttag
     3481 gggctaatac cattaaagaa cctggtcttt gcccatctgt gtcttctaat gaccaacaag
     3541 taccttcggc tgttcgttac aacgggtcaa agagagtgaa acctgaggaa gaagaagaga
     3601 gagacatgaa gaaatctagg ggattcgatg aaagggagtt gttttcgact gctgaatctt
     3661 cttcttcttc gagtgtgttt ttcgtttcgc agtcttgctc gttggcatca gaagggaaga
     3721 atctggaagg tattcaagat tcatctgatc agattactac aagtttggga aaaaatggtt
     3781 gcaaaggtgg cggtggctct ggaggtggtg ggtccggagg aggcggccgc atgaagctac
     3841 tgtcttctat cgaacaagca tgcgatattt gccgacttaa aaagctcaag tgctccaaag
     3901 aaaaaccgaa gtgcgccaag tgtctgaaga acaactggga gtgtcgctac tctcccaaaa
     3961 ccaaaaggtc tccgctgact agggcacatc tgacagaagt ggaatcaagg ctagaaagac
     4021 tggaatctag gggatccacc ggatctagat aactgatcat aatcagccat accacatttg
     4081 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     4141 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     4201 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4261 ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc
     4321 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     4381 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     4441 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     4501 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     4561 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     4621 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     4681 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     4741 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4801 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4861 tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     4921 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4981 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5041 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     5101 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     5161 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     5221 taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     5281 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     5341 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     5401 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca
     5461 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     5521 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     5581 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     5641 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     5701 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     5761 ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc
     5821 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     5881 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5941 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     6001 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     6061 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     6121 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     6181 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg
     6241 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     6301 taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt
     6361 cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg
     6421 tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca
     6481 acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt tagattgatt
     6541 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     6601 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     6661 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6721 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6781 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     6841 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6901 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6961 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     7021 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     7081 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     7141 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     7201 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     7261 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     7321 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
