LOCUS       sequence-348225-        8485 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cry2PHR with C-terminal IBAR  and WH2 domains.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8485)
  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 8485)
  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: Cry2PHR-IBAR-WH2
FEATURES             Location/Qualifiers
     source          1..8485
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..1494
                     /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      1513..1522
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1519..2226
                     /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(1664..1682)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1932..1951
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4649..4770
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4686..4705)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4740..4759
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4777..5232)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4914..4935)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5126..5145
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5259..5363
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5365..5722
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5365..5556
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5388..5408)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5573..5708
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5757..6551
                     /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             5757..6551
                     /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(5811..5830)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6421..6440
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6739..6758)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6783..6830
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7159..7747
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7648..7667
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7922..8225
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8226..8429
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8380..8400
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 atgaagatgg acaaaaagac tatagtttgg tttagaagag acctaaggat tgaggataat
       61 cctgcattag cagcagctgc tcacgaagga tctgtttttc ctgtcttcat ttggtgtcct
      121 gaagaagaag gacagtttta tcctggaaga gcttcaagat ggtggatgaa acaatcactt
      181 gctcacttat ctcaatcctt gaaggctctt ggatctgacc tcactttaat caaaacccac
      241 aacacgattt cagcgatctt ggattgtatc cgcgttaccg gtgctacaaa agtcgtcttt
      301 aaccacctct atgatcctgt ttcgttagtt cgggaccata ccgtaaagga gaagctggtg
      361 gaacgtggga tctctgtgca aagctacaat ggagatctat tgtatgaacc gtgggagata
      421 tactgcgaaa agggcaaacc ttttacgagt ttcaattctt actggaagaa atgcttagat
      481 atgtcgattg aatccgttat gcttcctcct ccttggcggt tgatgccaat aactgcagcg
      541 gctgaagcga tttgggcgtg ttcgattgaa gaactagggc tggagaatga ggccgagaaa
      601 ccgagcaatg cgttgttaac tagagcttgg tctccaggat ggagcaatgc tgataagtta
      661 ctaaatgagt tcatcgagaa gcagttgata gattatgcaa agaacagcaa gaaagttgtt
      721 gggaattcta cttcactact ttctccgtat ctccatttcg gggaaataag cgtcagacac
      781 gttttccagt gtgcccggat gaaacaaatt atatgggcaa gagataagaa cagtgaagga
      841 gaagaaagtg cagatctttt tcttagggga atcggtttaa gagagtattc tcggtatata
      901 tgtttcaact tcccgtttac tcacgagcaa tcgttgttga gtcatcttcg gtttttccct
      961 tgggatgctg atgttgataa gttcaaggcc tggagacaag gcaggaccgg ttatccgttg
     1021 gtggatgccg gaatgagaga gctttgggct accggatgga tgcataacag aataagagtg
     1081 attgtttcaa gctttgctgt gaagtttctt ctccttccat ggaaatgggg aatgaagtat
     1141 ttctgggata cacttttgga tgctgatttg gaatgtgaca tccttggctg gcagtatatc
     1201 tctgggagta tccccgatgg ccacgagctt gatcgcttgg acaatcccgc gttacaaggc
     1261 gccaaatatg acccagaagg tgagtacata aggcaatggc ttcccgagct tgcgagattg
     1321 ccaactgaat ggatccatca tccatgggac gctcctttaa ccgtactcaa agcttctggt
     1381 gtggaactcg gaacaaacta tgcgaaaccc attgtagaca tcgacacagc tcgtgagcta
     1441 ctagctaaag ctatttcaag aacccgtgaa gcacagatca tgatcggagc agcagcccgg
     1501 gatccaccgg tcgccaccat ggtgagcaag ggcgaggagg ataacatggc catcatcaag
     1561 gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc
     1621 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
     1681 aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc
     1741 aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag
     1801 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag
     1861 gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc
     1921 ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg
     1981 atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac
     2041 ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg
     2101 cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc
     2161 atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg
     2221 tacaagggcg gcagcggcgg cagcatggag gctgtgattg agaaggaatg cagcgcgctc
     2281 ggaggcctct tccagaccat catcagcgac atgaagggga gctatccagt ttgggaagat
     2341 ttcataaaca aagcaggaaa gctgcagtcc cagcttcgga caacagtagt agcagcagct
     2401 gccttcttgg acgcctttca gaaagtggct gacatggcca ccaacacacg tggtgggacc
     2461 agggagattg gatctgctct caccaggatg tgcatgaggc acagaagcat tgaagccaag
     2521 ctgaggcagt tttcgagcgc tttaattgat tgtctgataa acccacttca agaacagatg
     2581 gaagaatgga agaaagtggc caaccagctg gataaagacc acgcaaaaga atataagaaa
     2641 gcccgccaag agataaaaaa gaagtcctcg gatacgctga aactgcagaa gaaagcaaaa
     2701 aaagtggaca ctcttgggag aggtgatatc cagcctcagt tggacagtgc tctccaagat
     2761 gtcaatgata agtatctctt attggaagaa acagaaaagc aggctgtccg gaaggctttg
     2821 attgaagaac gtggccgatt ctgtaccttc atctctatgc tgcggccagt gattgaagaa
     2881 gaaatctcaa tgctagggga aataacccac cttcagacca tctcggaaga tctaaaaagc
     2941 ctgaccatgg accctcacaa actgccctcc tcaagtgaac aggtgattct ggacttgaaa
     3001 ggttctgatt acagctggtc gtatcagacg ccaccctctt cccccagcac caccatgtcc
     3061 agaaagtcca gtgtctgcag cagcctgaac agtgtcaaca gcagtgactc ccggtccagc
     3121 ggctcccact cgcattcccc cagctcacat taccgctacc gcagctccaa cctggcccag
     3181 caggctcctg tgaggctgtc cagcgtgtcc tcccatgact caggattcat atcccaggat
     3241 gccttccagt ccaagtcacc atcccccatg ccgccagagg cccccaacca gttgtctaac
     3301 gggttttctc actatagttt atcaagtgag tcccacgtgg ggcccacggg tgcaggcctt
     3361 ttccctcatt gcctgcctgc ctctcgcctg ctccctcggg tcacctctgt ccaccttcca
     3421 gactacgctc attattacac cattgggccc ggcatgttcc cgtcatctca gatccctagc
     3481 tggaaggact gggctaagcc tgggccctat gaccagcctc tggtgaacac cctgcagcgc
     3541 cgcaaagaga agcgagaacc ggaccccaac gggggaggac ccactaccgc cagcggccca
     3601 cccgcagcag ctgaggaggc tcagaggcca cggagcatga ctgtatcggc tgccaccagg
     3661 cctggtgagg agatggaggc ttgtgaggag ctggccctgg ccctgtctcg gggcctgcag
     3721 ctggacaccc agaggagcag ccgggactcg cttcagtgct ccagcggcta cagcacccag
     3781 acaaccaccc cctgctgctc tgaggacacc atcccttccc aagtttcaga ttatgattat
     3841 ttctctgtaa gtggtgacca ggaggcagat cagcaggagt tcgacaagtc ctccaccatt
     3901 ccaagaaaca gcgacatcag ccagtcctac cgacggatgt tccaagccaa gcgtccagcc
     3961 tcaactgctg gcctccccac caccctggga cctgctatgg tcactccagg ggttgcaact
     4021 atccgacgga ccccttccac caagccttct gtccgccggg gaaccattgg agctggtccc
     4081 atccccatca agacacccgt gatccctgtc aagaccccaa ccgtcccaga cctcccaggg
     4141 gtgttgccag cccctccaga tgggccagaa gagcgcgggg agcacagccc tgagtcgcca
     4201 tctgtgggtg agggccccca aggtgtcacc agcatgccct cctcaatgtg gagcggccaa
     4261 gcttccgtta accctccact tccaggcccg aagcccagta tccctgagga gcacagacag
     4321 gcaattccag aaagtgaagc tgaagaccag gaacgggaac ccccaagtgc cactgtctcc
     4381 ccaggccaga ttccagagag tgaccctgca gacctgagcc caagggatac tccacaagga
     4441 gaagacatgc tgaacgccat ccgaaggggc gtgaaactga agaagaccac gacaaacgat
     4501 cgctcagccc ctcgcttttc ttgataagcg gccgcgactc tagatcataa tcagccatac
     4561 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     4621 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     4681 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4741 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt
     4801 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     4861 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     4921 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     4981 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     5041 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     5101 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     5161 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     5221 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5281 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5341 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     5401 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     5461 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     5521 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     5581 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     5641 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     5701 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     5761 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     5821 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     5881 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     5941 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     6001 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     6061 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     6121 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     6181 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     6241 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     6301 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     6361 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     6421 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     6481 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     6541 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     6601 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     6661 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     6721 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     6781 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     6841 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     6901 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     6961 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     7021 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     7081 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     7141 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     7201 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     7261 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     7321 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     7381 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     7441 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     7501 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     7561 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     7621 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     7681 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     7741 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     7801 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     7861 ttagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     7921 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     7981 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     8041 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     8101 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     8161 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     8221 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     8281 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     8341 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     8401 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct
     8461 accggactca gatctcgagg ccacc
//
