LOCUS       sequence-254060-        6828 bp    DNA     circular SYN 15-APR-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 6828)
  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 6828)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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..6828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          620..1640
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1560..1582
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1648..1667
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3842..3861)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3907..3962
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3961..3980)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4323..4339)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4323..4339)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4336..4358)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4347..4363
                     /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(4371..4377,4378..4395,4396..4401))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4416..4437
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4496..4691
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4542..4677
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4697..4831
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4734..4753)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4788..4807
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4899..4916)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5070..5658)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5150..5169)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5829..6620,6621..6689))
                     /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     6452..6471
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6690..6794)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggggggg gcggggcgag gcggagaggt gcggcggcag ccaatcagag
      541 cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa
      601 gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc
      661 cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg
      721 gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc
      781 ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag ggccctttgt gcggggggag
      841 cggctcgggg ggtgcgtgcg tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct
      901 gcccggcggc tgtgagcgct gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg
      961 cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa
     1021 aggctgcgtg cggggtgtgt gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg
     1081 gctgcaaccc cccctgcacc cccctccccg agttgctgag cacggcccgg cttcgggtgc
     1141 ggggctccgt acggggcgtg gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt
     1201 gggggtgccg ggcggggcgg ggccgcctcg ggccggggag ggctcggggg aggggcgcgg
     1261 cggcccccgg agcgccggcg gctgtcgagg cgcggcgagc cgcagccatt gccttttatg
     1321 gtaatcgtgc gagagggcgc agggacttcc tttgtcccaa atctgtgcgg agccgaaatc
     1381 tgggaggcgc cgccgcaccc cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag
     1441 gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct
     1501 ccagcctcgg ggctgtccgc ggggggacgg ctgccttcgg gggggacggg gcagggcggg
     1561 gttcggcttc tggcgtgtga ccggcggctc tagctagagc ctctgctaac catgttcatg
     1621 ccttcttctt tttcctacag ctcctgggca acgtgctggt tattgtgctg tctcatcatt
     1681 ttggcaagaa ttcaccatga agatggacaa gaaaaccatc gtgtggtttc ggagagatct
     1741 gaggattgag gacaacccag ccctggccgc tgccgctcac gaaggcagcg tgtttccagt
     1801 gttcatctgg tgccccgagg aagagggaca gttctaccca ggaagggcca gccgctggtg
     1861 gatgaagcag tccctggctc acctgtctca gagtctgaag gccctggggt cagatctgac
     1921 cctgatcaaa acccataaca caatcagcgc cattctggac tgtattcggg tgactggagc
     1981 caccaaagtg gtgtttaatc acctgtatga tcccgtgagc ctggtgcggg accatacagt
     2041 gaaggagaaa ctggtggaaa gaggcatcag tgtgcagtca tacaacgggg atctgctgta
     2101 tgagccatgg gaaatctact gcgagaaggg gaaacccttt accagcttca attcctactg
     2161 gaagaaatgt ctggacatga gcgagctgat taaggagaac atgcacatga agctgtacat
     2221 ggagggcacc gtggacaacc atcacttcaa gtgcacatcc gagggcgaag gcaagcccta
     2281 cgagggcacc cagaccatga gaatcaaggt ggtcgagggc ggccctctcc ccttcgcctt
     2341 cgacatcctg gctactagct tcctctacgg cagcaagacc ttcatcaacc acacccaggg
     2401 catccccgac ttcttcaagc agtccttccc tgagggcttc acatgggaga gagtcaccac
     2461 atacgaagac gggggcgtgc tgaccgctac ccaggacacc agcctccagg acggctgcct
     2521 catctacaac gtcaagatca gaggggtgaa cttcacatcc aacggccctg tgatgcagaa
     2581 gaaaacactc ggctgggagg cattcaccga gacgctgtac cccgctgacg gcggcctgga
     2641 aggcagaaac gacatggccc tgaagctcgt gggcgggagc catctgatcg caaacgccaa
     2701 gaccacatat agatccaaga aacccgctaa gaacctcaag atgcctggcg tctactatgt
     2761 ggactacaga ctggaaagaa tcaaggaggc caacaacgag acctacgtcg agcagcacga
     2821 ggtggcagtg gccagatacc catggaggct gatgcctatt acagccgctg ccgaagctat
     2881 ctgggcctgc tccattgaag agctgggact ggaaaacgag gccgaaaagc cttctaatgc
     2941 tctgctgact cgcgcctggt ccccaggatg gtctaacgct gataaactgc tgaatgagtt
     3001 catcgaaaag cagctgattg actatgccaa aaactctaag aaagtggtgg gcaacagcac
     3061 cagcctgctg tctccatacc tgcactttgg agagatcagt gtgcggcacg tgttccagtg
     3121 cgctagaatg aagcagatca tttgggccag ggataaaaac agtgagggcg aagagtcagc
     3181 cgacctgttt ctgaggggaa tcggactgag ggagtactcc cgctatattt gtttcaattt
     3241 tcccttcact cacgaacagt cactgctgag ccatctgcgg ttctttcctt gggacgctga
     3301 tgtggacaag ttcaaagcct ggcggcaggg aagaacaggc tatcccctgg tggatgctgg
     3361 aatgagagag ctgtgggcca ctggctggat gcacaatagg atccgcgtga ttgtgagctc
     3421 ctttgctgtg aagttcctgc tgctgccttg gaagtggggc atgaaatatt tttgggatac
     3481 actgctagct gccgacctgg agtgtgacat cctggggtgg cagtacatct ccggatctat
     3541 tccagatggc catgaactgg ataggctgga caaccccgct ctgcagggcg ctaagtatga
     3601 ccctgaggga gaatacatca ggcagtggct gcctgagctg gctaggctgc caactgaatg
     3661 gattcaccat ccctgggacg ctcctctgac agtgctgaag gccagtgggg tggagctggg
     3721 aactaattac gctaaaccta tcgtggatat tgacaccgcc agagagctgc tggctaaggc
     3781 catctcccgg accagagaag cccagatcat gattggcgct gcctctagtt aagtcgacgg
     3841 atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt
     3901 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct
     3961 cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt
     4021 agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag
     4081 gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga
     4141 cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa
     4201 aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca
     4261 tagctgtccc tcttctctta tggagatccc tcgacctgca gcccaagctt ggcgtaatca
     4321 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     4381 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     4441 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc
     4501 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4561 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4621 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4681 gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4741 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4801 actcatcaat gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg
     4861 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4921 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4981 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5041 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5101 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5161 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5221 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5281 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5341 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5401 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5461 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     5521 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5581 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5641 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5701 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5761 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5821 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5881 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5941 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6001 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6061 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6121 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6181 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6241 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6301 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6361 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6421 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6481 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6541 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6601 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6661 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6721 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6781 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg ggtcgatc
//
