LOCUS       sequence-355865-        7419 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cry2PHR with N-terminal IBAR domain and C-terminal WH2 domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7419)
  AUTHORS   Wurz AI, Bunner WP, Szatmari EM, Hughes RM
  TITLE     CRY-BARs: Versatile light-gated molecular tools for the remodeling 
            of membrane architectures.
  JOURNAL   J Biol Chem. 2022 Aug 18;298(10):102388. doi: 
            10.1016/j.jbc.2022.102388.
  PUBMED    35987384
REFERENCE   2  (bases 1 to 7419)
  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: "2022-08-18"; volume: "298(10):102388. doi"; pages: " 
            10.1016/j.jbc.2022.102388"
COMMENT     Sequence Label: IBAR-Cry2PHR-WH2
FEATURES             Location/Qualifiers
     source          1..7419
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1299..2792
                     /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      2811..2820
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2817..3524
                     /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(2962..2980)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3230..3249
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4147..4268
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4184..4203)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4238..4257
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4275..4730)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4412..4433)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4624..4643
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4757..4861
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4863..5220
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4863..5054
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4886..4906)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5071..5206
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5255..6049
                     /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             5255..6049
                     /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(5309..5328)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5919..5938
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6237..6256)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6281..6328
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6657..7245
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7146..7165
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcatgg
      541 aggctgtgat tgagaaggaa tgcagcgcgc tcggaggcct cttccagacc atcatcagcg
      601 acatgaaggg gagctatcca gtttgggaag atttcataaa caaagcagga aagctgcagt
      661 cccagcttcg gacaacagta gtagcagcag ctgccttctt ggacgccttt cagaaagtgg
      721 ctgacatggc caccaacaca cgtggtggga ccagggagat tggatctgct ctcaccagga
      781 tgtgcatgag gcacagaagc attgaagcca agctgaggca gttttcgagc gctttaattg
      841 attgtctgat aaacccactt caagaacaga tggaagaatg gaagaaagtg gccaaccagc
      901 tggataaaga ccacgcaaaa gaatataaga aagcccgcca agagataaaa aagaagtcct
      961 cggatacgct gaaactgcag aagaaagcaa aaaaagtgga cactcttggg agaggtgata
     1021 tccagcctca gttggacagt gctctccaag atgtcaatga taagtatctc ttattggaag
     1081 aaacagaaaa gcaggctgtc cggaaggctt tgattgaaga acgtggccga ttctgtacct
     1141 tcatctctat gctgcggcca gtgattgaag aagaaatctc aatgctaggg gaaataaccc
     1201 accttcagac catctcggaa gatctaaaaa gcctgaccat ggaccctcac aaactgccct
     1261 cctcaagtga acaggtgatt ctggacctcg aggccaccat gaagatggac aaaaagacta
     1321 tagtttggtt tagaagagac ctaaggattg aggataatcc tgcattagca gcagctgctc
     1381 acgaaggatc tgtttttcct gtcttcattt ggtgtcctga agaagaagga cagttttatc
     1441 ctggaagagc ttcaagatgg tggatgaaac aatcacttgc tcacttatct caatccttga
     1501 aggctcttgg atctgacctc actttaatca aaacccacaa cacgatttca gcgatcttgg
     1561 attgtatccg cgttaccggt gctacaaaag tcgtctttaa ccacctctat gatcctgttt
     1621 cgttagttcg ggaccatacc gtaaaggaga agctggtgga acgtgggatc tctgtgcaaa
     1681 gctacaatgg agatctattg tatgaaccgt gggagatata ctgcgaaaag ggcaaacctt
     1741 ttacgagttt caattcttac tggaagaaat gcttagatat gtcgattgaa tccgttatgc
     1801 ttcctcctcc ttggcggttg atgccaataa ctgcagcggc tgaagcgatt tgggcgtgtt
     1861 cgattgaaga actagggctg gagaatgagg ccgagaaacc gagcaatgcg ttgttaacta
     1921 gagcttggtc tccaggatgg agcaatgctg ataagttact aaatgagttc atcgagaagc
     1981 agttgataga ttatgcaaag aacagcaaga aagttgttgg gaattctact tcactacttt
     2041 ctccgtatct ccatttcggg gaaataagcg tcagacacgt tttccagtgt gcccggatga
     2101 aacaaattat atgggcaaga gataagaaca gtgaaggaga agaaagtgca gatctttttc
     2161 ttaggggaat cggtttaaga gagtattctc ggtatatatg tttcaacttc ccgtttactc
     2221 acgagcaatc gttgttgagt catcttcggt ttttcccttg ggatgctgat gttgataagt
     2281 tcaaggcctg gagacaaggc aggaccggtt atccgttggt ggatgccgga atgagagagc
     2341 tttgggctac cggatggatg cataacagaa taagagtgat tgtttcaagc tttgctgtga
     2401 agtttcttct ccttccatgg aaatggggaa tgaagtattt ctgggataca cttttggatg
     2461 ctgatttgga atgtgacatc cttggctggc agtatatctc tgggagtatc cccgatggcc
     2521 acgagcttga tcgcttggac aatcccgcgt tacaaggcgc caaatatgac ccagaaggtg
     2581 agtacataag gcaatggctt cccgagcttg cgagattgcc aactgaatgg atccatcatc
     2641 catgggacgc tcctttaacc gtactcaaag cttctggtgt ggaactcgga acaaactatg
     2701 cgaaacccat tgtagacatc gacacagctc gtgagctact agctaaagct atttcaagaa
     2761 cccgtgaagc acagatcatg atcggagcag cagcccggga tccaccggtc gccaccatgg
     2821 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc
     2881 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     2941 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
     3001 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
     3061 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     3121 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg
     3181 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
     3241 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
     3301 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
     3361 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
     3421 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
     3481 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagggcggc agcggcggca
     3541 gcccttctgt ccgccgggga accattggag ctggtcccat ccccatcaag acacccgtga
     3601 tccctgtcaa gaccccaacc gtcccagacc tcccaggggt gttgccagcc cctccagatg
     3661 ggccagaaga gcgcggggag cacagccctg agtcgccatc tgtgggtgag ggcccccaag
     3721 gtgtcaccag catgccctcc tcaatgtgga gcggccaagc ttccgttaac cctccacttc
     3781 caggcccgaa gcccagtatc cctgaggagc acagacaggc aattccagaa agtgaagctg
     3841 aagaccagga acgggaaccc ccaagtgcca ctgtctcccc aggccagatt ccagagagtg
     3901 accctgcaga cctgagccca agggatactc cacaaggaga agacatgctg aacgccatcc
     3961 gaaggggcgt gaaactgaag aagaccacga caaacgatcg ctcagcccct cgcttttctt
     4021 gataagcggc cgcgactcta gatcataatc agccatacca catttgtaga ggttttactt
     4081 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     4141 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4201 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4261 gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     4321 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     4381 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     4441 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     4501 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     4561 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     4621 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     4681 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
     4741 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4801 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4861 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4921 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4981 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5041 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     5101 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
     5161 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     5221 gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     5281 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     5341 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     5401 caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc ggctatcgtg
     5461 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     5521 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     5581 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     5641 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     5701 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     5761 actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg
     5821 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     5881 tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc
     5941 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     6001 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg
     6061 gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     6121 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     6181 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag gctaactgaa
     6241 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     6301 aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     6361 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
     6421 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     6481 ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa acttcatttt
     6541 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6601 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6661 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6721 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6781 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6841 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6901 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6961 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     7021 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     7081 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7141 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7201 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     7261 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     7321 tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttattaat agtaatcaat
     7381 tacggggtca ttagttcata gcccatatat ggagttccg
//
