LOCUS       sequence-263360-        7188 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of GFP-tagged CRY2PHR fused to inositol 5-phosphatase 
            domain of OCRL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7188)
  AUTHORS   Idevall-Hagren O, Dickson EJ, Hille B, Toomre DK, De Camilli P
  TITLE     Optogenetic control of phosphoinositide metabolism.
  JOURNAL   Proc Natl Acad Sci U S A. 2012 Aug 28;109(35):E2316-23. doi: 
            10.1073/pnas.1211305109. Epub 2012 Jul 30.
  PUBMED    22847441
REFERENCE   2  (bases 1 to 7188)
  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: 
            "10.1073/pnas.1211305109"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2012-08-28- 28"; volume: "109"; issue: "35"; pages: 
            "E2316-23"
COMMENT     Sequence Label: GFP-CRY2-OCRL
FEATURES             Location/Qualifiers
     source          1..7188
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        69..372
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        373..576
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     527..547
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      615..624
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(621..623,624..626,627..1337)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(621..623,624..626,627..1337)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(666..687)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(927..946)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1274..1295
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1383..2876
                     /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"
     polyA_signal    3984..4105
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4021..4040)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4075..4094
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4112..4567)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4249..4270)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4461..4480
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4594..4698
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4700..5057
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4700..4891
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4723..4743)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4908..5043
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5092..5886
                     /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             5092..5886
                     /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(5146..5165)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5756..5775
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6074..6093)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6118..6165
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6494..7082
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6983..7002
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
       61 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      121 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      181 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      241 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      301 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      361 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      421 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      481 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      541 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc
      601 tagcgctacc ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
      661 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
      721 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
      781 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
      841 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
      901 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
      961 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     1021 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     1081 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     1141 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     1201 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     1261 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     1321 tggacgagct gtacaagaga tctcgaagcg cggccgcggg agcaggagga gcagctcgag
     1381 ctatgaagat ggacaaaaag actatagttt ggtttagaag agacctaagg attgaggata
     1441 atcctgcatt agcagcagct gctcacgaag gatctgtttt tcctgtcttc atttggtgtc
     1501 ctgaagaaga aggacagttt tatcctggaa gagcttcaag atggtggatg aaacaatcac
     1561 ttgctcactt atctcaatcc ttgaaggctc ttggatctga cctcacttta atcaaaaccc
     1621 acaacacgat ttcagcgatc ttggattgta tccgcgttac cggtgctaca aaagtcgtct
     1681 ttaaccacct ctatgatcct gtttcgttag ttcgggacca taccgtaaag gagaagctgg
     1741 tggaacgtgg gatctctgtg caaagctaca atggagatct attgtatgaa ccgtgggaga
     1801 tatactgcga aaagggcaaa ccttttacga gtttcaattc ttactggaag aaatgcttag
     1861 atatgtcgat tgaatccgtt atgcttcctc ctccttggcg gttgatgcca ataactgcag
     1921 cggctgaagc gatttgggcg tgttcgattg aagaactagg gctggagaat gaggccgaga
     1981 aaccgagcaa tgcgttgtta actagagctt ggtctccagg atggagcaat gctgataagt
     2041 tactaaatga gttcatcgag aagcagttga tagattatgc aaagaacagc aagaaagttg
     2101 ttgggaattc tacttcacta ctttctccgt atctccattt cggggaaata agcgtcagac
     2161 acgttttcca gtgtgcccgg atgaaacaaa ttatatgggc aagagataag aacagtgaag
     2221 gagaagaaag tgcagatctt tttcttaggg gaatcggttt aagagagtat tctcggtata
     2281 tatgtttcaa cttcccgttt actcacgagc aatcgttgtt gagtcatctt cggtttttcc
     2341 cttgggatgc tgatgttgat aagttcaagg cctggagaca aggcaggacc ggttatccgt
     2401 tggtggatgc cggaatgaga gagctttggg ctaccggatg gatgcataac agaataagag
     2461 tgattgtttc aagctttgct gtgaagtttc ttctccttcc atggaaatgg ggaatgaagt
     2521 atttctggga tacacttttg gatgctgatt tggaatgtga catccttggc tggcagtata
     2581 tctctgggag tatccccgat ggccacgagc ttgatcgctt ggacaatccc gcgttacaag
     2641 gcgccaaata tgacccagaa ggtgagtaca taaggcaatg gcttcccgag cttgcgagat
     2701 tgccaactga atggatccat catccatggg acgctccttt aaccgtactc aaagcttctg
     2761 gtgtggaact cggaacaaac tatgcgaaac ccattgtaga catcgacaca gctcgtgagc
     2821 tactagctaa agctatttca agaacccgtg aagcacagat catgatcgga gcagcagctc
     2881 gaggcgcagc agcgggagca ggaggagcag gtcgatcgta tgtcaacatt cagactttca
     2941 gattttttgt tggaacttgg aatgtgaatg gccagtctcc agatagcggg ttagaacctt
     3001 ggctgaactg tgatcccaat cctcctgata tctactgcat tggattccaa gaactggact
     3061 tgagcacaga agccttcttc tactttgaat ctgtgaagga acaagaatgg tccatggctg
     3121 tagagagagg tttgcattcc aaagccaagt ataagaaagt tcaactggtg cgccttgttg
     3181 ggatgatgct tcttatattt gccagaaagg atcagtgtcg atacattcgt gatattgcta
     3241 cagaaacagt tggaactgga atcatgggga aaatgggaaa caaaggtggg gtagctgtga
     3301 gatttgtatt tcacaacacc accttttgca ttgtcaattc ccatctggct gcacacgtgg
     3361 aggactttga gagaaggaat caagattata aggacatttg tgcgagaatg agttttgtgg
     3421 tcccaaatca gaccctcccg cagttgaaca tcatgaaaca tgaggttgtc atttggttgg
     3481 gagatttgaa ttatagactt tgcatgcctg atgccaatga ggtgaaaagt cttattaata
     3541 agaaagacct tcagagactc ttgaaattcg accagctaaa tattcagcgc acacagaaaa
     3601 aagcttttgt tgacttcaat gaaggggaaa tcaagttcat ccccacttat aagtatgact
     3661 ctaaaacaga ccggtgggat tccagtggga aatgccgggt tccagcctgg tgtgaccgaa
     3721 ttctttggag aggaacaaat gttaatcagc ttaattatcg gagtcacatg gaactgaaaa
     3781 ccagcgacca caagcctgtt agcgccctct tccatattgg ggtgaaggtt gtggattgag
     3841 gtaccgcggg cccgggatcc accggatcta gataactgat cataatcagc cataccacat
     3901 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     3961 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     4021 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4081 tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa
     4141 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     4201 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     4261 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     4321 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     4381 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     4441 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     4501 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     4561 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4621 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4681 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     4741 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4801 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4861 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4921 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4981 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     5041 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     5101 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     5161 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     5221 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     5281 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     5341 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     5401 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     5461 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     5521 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5581 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     5641 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5701 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5761 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5821 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5881 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5941 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     6001 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     6061 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     6121 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     6181 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     6241 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     6301 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     6361 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     6421 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6481 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6541 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6601 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6661 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6721 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6781 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6841 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6901 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6961 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     7021 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     7081 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     7141 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccg
//
