LOCUS       sequence-248891-        6874 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  2P-activatable CRY2 with the FAD-disrupting mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6874)
  AUTHORS   Kinjo T, Terai K, Horita S, Nomura N, Sumiyama K, Togashi K, Iwata 
            S, Matsuda M
  TITLE     FRET-assisted photoactivation of flavoproteins for in vivo 
            two-photon optogenetics.
  JOURNAL   Nat Methods. 2019 Sep 9. pii: 10.1038/s41592-019-0541-5. doi: 
            10.1038/s41592-019-0541-5.
  PUBMED    31501546
REFERENCE   2  (bases 1 to 6874)
  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: "Nat 
            Methods. 2019 Sep 9. pii: 10.1038/s41592-019-0541-5. doi: 
            10.1038/s41592-019-0541-5."
COMMENT     Sequence Label: pCAGGS_2paCRY2(D387A)
FEATURES             Location/Qualifiers
     source          1..6874
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        8..387
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..665
                     /label=chicken beta-actin promoter
     intron          666..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1598..1620
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     complement(3898..3917)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3963..4018
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4017..4036)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4377..4393)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4377..4393)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4390..4412)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4401..4417
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4425..4431,4432..4449,4450..4455))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4470..4491
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4550..4745
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4596..4731
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4751..4885
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4788..4807)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4842..4861
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4953..4970)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5124..5712)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5204..5223)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5883..6674,6675..6743))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6506..6525
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6744..6848)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgatcgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
       61 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      121 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      181 atagggactt tccattgacg tcaatgggtg gactatttac ggtaaactgc ccacttggca
      241 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      301 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      361 tacgtattag tcatcgctat taccatgggt cgaggtgagc cccacgttct gcttcactct
      421 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
      481 tgcagcgatg ggggcggggg gggggggggc gcgcgccagg cggggcgggg cgggggcgag
      541 ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa
      601 agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc
      661 gggcgggagt cgctgcgttg ccttcgcccc gtgccccgct ccgcgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggctcgt ttcttttctg tggctgcgtg
      841 aaagccttaa agggctccgg gagggccctt tgtgcggggg ggagcggctc ggggggtgcg
      901 tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggcccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcgtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
     1081 tgcgtggggg ggtgagcagg gggtgtgggc gcggcggtcg ggctgtaacc cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtgcggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccg gagcgccggc
     1321 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
     1381 cagggacttc ctttgtccca aatctggcgg agccgaaatc tgggaggcgc cgccgcaccc
     1441 cctctagcgg gcgcgggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg
     1501 ccttcgtgcg tcgccgcgcc gccgtcccct tctccatctc cagcctcggg gctgccgcag
     1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1621 ggcggctcta gctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt ggcaagaatt caccatgaag
     1741 atggacaaga aaaccatcgt gtggtttcgg agagatctga ggattgagga caacccagcc
     1801 ctggccgctg ccgctcacga aggcagcgtg tttccagtgt tcatctggtg ccccgaggaa
     1861 gagggacagt tctacccagg aagggccagc cgctggtgga tgaagcagtc cctggctcac
     1921 ctgtctcaga gtctgaaggc cctggggtca gatctgaccc tgatcaaaac ccataacaca
     1981 atcagcgcca ttctggactg tattcgggtg actggagcca ccaaagtggt gtttaatcac
     2041 ctgtatgatc ccgtgagcct ggtgcgggac catacagtga aggagaaact ggtggaaaga
     2101 ggcatcagtg tgcagtcata caacggggat ctgctgtatg agccatggga aatctactgc
     2161 gagaagggga aaccctttac cagcttcaat tcctactgga agaaatgtct ggacggatct
     2221 ggaatgagcg agctgattaa ggagaacatg cacatgaagc tgtacatgga gggcaccgtg
     2281 gacaaccatc acttcaagtg cacatccgag ggcgaaggca agccctacga gggcacccag
     2341 accatgagaa tcaaggtggt cgagggcggc cctctcccct tcgccttcga catcctggct
     2401 actagcttcc tctacggcag caagaccttc atcaaccaca cccagggcat ccccgacttc
     2461 ttcaagcagt ccttccctga gggcttcaca tgggagagag tcaccacata cgaagacggg
     2521 ggcgtgctga ccgctaccca ggacaccagc ctccaggacg gctgcctcat ctacaacgtc
     2581 aagatcagag gggtgaactt cacatccaac ggccctgtga tgcagaagaa aacactcggc
     2641 tgggaggcat tcaccgagac gctgtacccc gctgacggcg gcctggaagg cagaaacgac
     2701 atggccctga agctcgtggg cgggagccat ctgatcgcaa acgccaagac cacatataga
     2761 tccaagaaac ccgctaagaa cctcaagatg cctggcgtct actatgtgga ctacagactg
     2821 gaaagaatca aggaggccaa caacgagacc tacgtcgagc agcacgaggt ggcagtggcc
     2881 agatacggta gcggtccatg gaggctgatg cctattacag ccgctgccga agctatctgg
     2941 gcctgctcca ttgaagagct gggactggaa aacgaggccg aaaagccttc taatgctctg
     3001 ctgactcgcg cctggtcccc aggatggtct aacgctgata aactgctgaa tgagttcatc
     3061 gaaaagcagc tgattgacta tgccaaaaac tctaagaaag tggtgggcaa cagcaccagc
     3121 ctgctgtctc catacctgca ctttggagag atcagtgtgc ggcacgtgtt ccagtgcgct
     3181 agaatgaagc agatcatttg ggccagggat aaaaacagtg agggcgaaga gtcagccgac
     3241 ctgtttctga ggggaatcgg actgagggag tactcccgct atatttgttt caattttccc
     3301 ttcactcacg aacagtcact gctgagccat ctgcggttct ttccttggga cgctgatgtg
     3361 gacaagttca aagcctggcg gcagggaaga acaggctatc ccctggtgga tgctggaatg
     3421 agagagctgt gggccactgg ctggatgcac aataggatcc gcgtgattgt gagctccttt
     3481 gctgtgaagt tcctgctgct gccttggaag tggggcatga aatatttttg ggatacactg
     3541 ctagctgccg acctggagtg tgacatcctg gggtggcagt acatctccgg atctattcca
     3601 gatggccatg aactggatag gctggacaac cccgctctgc agggcgctaa gtatgaccct
     3661 gagggagaat acatcaggca gtggctgcct gagctggcta ggctgccaac tgaatggatt
     3721 caccatccct gggacgctcc tctgacagtg ctgaaggcca gtggggtgga gctgggaact
     3781 aattacgcta aacctatcgt ggatattgac accgccagag agctgctggc taaggccatc
     3841 tcccggacca gagaagccca gatcatgatt ggcgctgcct ctagataagt cgacggatct
     3901 ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg
     3961 ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact
     4021 cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag
     4081 tttggcaaca tatgccatat gctggctgcc atgaacaaag gtggctataa agaggtcatc
     4141 agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag
     4201 gttagatttt tttttatatt ttgtttgtgt tatttttttc tttaacatcc ctaaaatttt
     4261 ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg
     4321 tccctcttct cttatgaaga tccctcgacc tgcagcccaa gcttggcgta atcatggtca
     4381 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4441 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4501 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt
     4561 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4621 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4681 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4741 gcaaaaagct aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4801 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4861 caatgtatct tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga
     4921 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4981 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5041 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5101 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5161 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5221 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5281 tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc
     5341 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5401 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5461 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5521 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     5581 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5641 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5701 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5761 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5821 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5881 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5941 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6001 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6061 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6121 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6181 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6241 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6301 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6361 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6421 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6481 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6541 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6601 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6661 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6721 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6781 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6841 gttccgcgca catttccccg aaaagtgcca cctg
//
