LOCUS       sequence-229778-        6877 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Light activated recruitment of BAX in mammalian cells with Cry2/CIB 
            optogenetic switch.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6877)
  AUTHORS   Godwin WC, Hoffmann GF, Gray TJ, Hughes RM
  TITLE     Imaging of morphological and biochemical hallmarks of apoptosis with
            optimized optogenetic tools.
  JOURNAL   J Biol Chem. 2019 Nov 8;294(45):16918-16929. doi: 
            10.1074/jbc.RA119.009141. Epub 2019 Oct 3.
  PUBMED    31582560
REFERENCE   2  (bases 1 to 6877)
  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: 
            "10.1074/jbc.RA119.009141"; journalName: "J Biol Chem"; date: 
            "2019-11-8- 8"; volume: "294"; issue: "45"; pages: "16918-16929"
COMMENT     Sequence Label: Cry2(1-531)-mCh-BAXS184E
FEATURES             Location/Qualifiers
     source          1..6877
                     /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"
     CDS             619..2112
                     /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      2230..2239
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2236..2943
                     /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(2381..2399)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2649..2668
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3665..3786
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3702..3721)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3756..3775
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3793..4248)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3930..3951)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4142..4161
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4275..4379
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4381..4738
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4381..4572
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4404..4424)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4589..4724
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4773..5567
                     /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             4773..5567
                     /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(4827..4846)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5437..5456
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5755..5774)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5799..5846
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6175..6763
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6664..6683
                     /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 ccggactcag atctcgagat gaagatggac aaaaagacta tagtttggtt tagaagagac
      661 ctaaggattg aggataatcc tgcattagca gcagctgctc acgaaggatc tgtttttcct
      721 gtcttcattt ggtgtcctga agaagaagga cagttttatc ctggaagagc ttcaagatgg
      781 tggatgaaac aatcacttgc tcacttatct caatccttga aggctcttgg atctgacctc
      841 actttaatca aaacccacaa cacgatttca gcgatcttgg attgtatccg cgttaccggt
      901 gctacaaaag tcgtctttaa ccacctctat gatcctgttt cgttagttcg ggaccatacc
      961 gtaaaggaga agctggtgga acgtgggatc tctgtgcaaa gctacaatgg agatctattg
     1021 tatgaaccgt gggagatata ctgcgaaaag ggcaaacctt ttacgagttt caattcttac
     1081 tggaagaaat gcttagatat gtcgattgaa tccgttatgc ttcctcctcc ttggcggttg
     1141 atgccaataa ctgcagcggc tgaagcgatt tgggcgtgtt cgattgaaga actagggctg
     1201 gagaatgagg ccgagaaacc gagcaatgcg ttgttaacta gagcttggtc tccaggatgg
     1261 agcaatgctg ataagttact aaatgagttc atcgagaagc agttgataga ttatgcaaag
     1321 aacagcaaga aagttgttgg gaattctact tcactacttt ctccgtatct ccatttcggg
     1381 gaaataagcg tcagacacgt tttccagtgt gcccggatga aacaaattat atgggcaaga
     1441 gataagaaca gtgaaggaga agaaagtgca gatctttttc ttaggggaat cggtttaaga
     1501 gagtattctc ggtatatatg tttcaacttc ccgtttactc acgagcaatc gttgttgagt
     1561 catcttcggt ttttcccttg ggatgctgat gttgataagt tcaaggcctg gagacaaggc
     1621 aggaccggtt atccgttggt ggatgccgga atgagagagc tttgggctac cggatggatg
     1681 cataacagaa taagagtgat tgtttcaagc tttgctgtga agtttcttct ccttccatgg
     1741 aaatggggaa tgaagtattt ctgggataca cttttggatg ctgatttgga atgtgacatc
     1801 cttggctggc agtatatctc tgggagtatc cccgatggcc acgagcttga tcgcttggac
     1861 aatcccgcgt tacaaggcgc caaatatgac ccagaaggtg agtacataag gcaatggctt
     1921 cccgagcttg cgagattgcc aactgaatgg atccatcatc catgggacgc tcctttaacc
     1981 gtactcaaag cttctggtgt ggaactcgga acaaactatg cgaaacccat tgtagacatc
     2041 gacacagctc gtgagctact agctaaagct atttcaagaa cccgtgaagc acagatcatg
     2101 atcggagcag cacctgatga gattgtagca gatagcttcg aggccttagg ggctaatacc
     2161 attaaagaac ctggtctttg cccatctgtg tcttctaatg accaacaagt agcccgggat
     2221 ccaccggtcg ccaccatggt gagcaagggc gaggaggata acatggccat catcaaggag
     2281 ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag
     2341 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
     2401 ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag
     2461 gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc
     2521 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac
     2581 tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc
     2641 tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg
     2701 taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc
     2761 ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc
     2821 ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc
     2881 gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac
     2941 aagggcggca gcggcggcgg catggacggg tccggggagc agcccagagg cggggggccc
     3001 accagctctg agcagatcat gaagacaggg gcccttttgc ttcagggttt catccaggat
     3061 cgagcagggc gaatgggggg ggaggcaccc gagctggccc tggacccggt gcctcaggat
     3121 gcgtccacca agaagctgag cgagtgtctc aagcgcatcg gggacgaact ggacagtaac
     3181 atggagctgc agaggatgat tgccgccgtg gacacagact ccccccgaga ggtctttttc
     3241 cgagtggcag ctgacatgtt ttctgacggc aacttcaact ggggccgggt tgtcgccctt
     3301 ttctactttg ccagcaaact ggtgctcaag gccctgtgca ccaaggtgcc ggaactgatc
     3361 agaaccatca tgggctggac attggacttc ctccgggagc ggctgttggg ctggatccaa
     3421 gaccagggtg gttgggacgg cctcctctcc tactttggga cgcccacgtg gcagaccgtg
     3481 accatctttg tggcgggagt gctcaccgcc gagctcacca tctggaagaa gatgggctaa
     3541 tgagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
     3601 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     3661 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3721 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3781 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     3841 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     3901 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     3961 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     4021 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     4081 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     4141 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     4201 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     4261 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4321 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     4381 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     4441 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     4501 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4561 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4621 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     4681 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     4741 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4801 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4861 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4921 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     4981 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5041 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5101 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5161 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     5221 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5281 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5341 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5401 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5461 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5521 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5581 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5641 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5701 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     5761 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     5821 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     5881 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     5941 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     6001 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     6061 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6121 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6181 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6241 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6301 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     6361 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6421 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6481 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6541 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6601 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6661 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6721 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6781 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6841 ccctgattct gtggataacc gtattaccgc catgcat
//
