LOCUS       sequence-254052-        6847 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  2P-activatable CRY2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6847)
  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 6847)
  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
FEATURES             Location/Qualifiers
     source          1..6847
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          621..1641
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1561..1583
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1649..1668
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3861..3880)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3926..3981
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3980..3999)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4342..4358)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4342..4358)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4355..4377)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4366..4382
                     /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(4390..4396,4397..4414,4415..4420))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4435..4456
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4515..4710
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4561..4696
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4716..4850
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4753..4772)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4807..4826
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4918..4935)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5089..5677)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5169..5188)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5848..6639,6640..6708))
                     /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     6471..6490
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6709..6813)
                     /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 ggcggggcga ggcggagagg tgcggcggca gccaatcaga
      541 gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa
      601 agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg
      661 ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg
      721 ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga cggcttgttt
      781 cttttctgtg gctgcgtgaa agccttgagg ggctccggga gggccctttg tgcgggggga
      841 gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc
      901 tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc
      961 gcgaggggag cgcggccggg ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca
     1021 aaggctgcgt gcggggtgtg tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg
     1081 ggctgcaacc ccccctgcac ccccctcccc gagttgctga gcacggcccg gcttcgggtg
     1141 cggggctccg tacggggcgt ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg
     1201 tgggggtgcc gggcggggcg gggccgcctc gggccgggga gggctcgggg gaggggcgcg
     1261 gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat tgccttttat
     1321 ggtaatcgtg cgagagggcg cagggacttc ctttgtccca aatctgtgcg gagccgaaat
     1381 ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca
     1441 ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc
     1501 tccagcctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg ggcagggcgg
     1561 ggttcggctt ctggcgtgtg accggcggct ctagctagag cctctgctaa ccatgttcat
     1621 gccttcttct ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat
     1681 tttggcaaga attcaccatg aagatggaca agaaaaccat cgtgtggttt cggagagatc
     1741 tgaggattga ggacaaccca gccctggccg ctgccgctca cgaaggcagc gtgtttccag
     1801 tgttcatctg gtgccccgag gaagagggac agttctaccc aggaagggcc agccgctggt
     1861 ggatgaagca gtccctggct cacctgtctc agagtctgaa ggccctgggg tcagatctga
     1921 ccctgatcaa aacccataac acaatcagcg ccattctgga ctgtattcgg gtgactggag
     1981 ccaccaaagt ggtgtttaat cacctgtatg atcccgtgag cctggtgcgg gaccatacag
     2041 tgaaggagaa actggtggaa agaggcatca gtgtgcagtc atacaacggg gatctgctgt
     2101 atgagccatg ggaaatctac tgcgagaagg ggaaaccctt taccagcttc aattcctact
     2161 ggaagaaatg tctggacgga tctggaatga gcgagctgat taaggagaac atgcacatga
     2221 agctgtacat ggagggcacc gtggacaacc atcacttcaa gtgcacatcc gagggcgaag
     2281 gcaagcccta cgagggcacc cagaccatga gaatcaaggt ggtcgagggc ggccctctcc
     2341 ccttcgcctt cgacatcctg gctactagct tcctctacgg cagcaagacc ttcatcaacc
     2401 acacccaggg catccccgac ttcttcaagc agtccttccc tgagggcttc acatgggaga
     2461 gagtcaccac atacgaagac gggggcgtgc tgaccgctac ccaggacacc agcctccagg
     2521 acggctgcct catctacaac gtcaagatca gaggggtgaa cttcacatcc aacggccctg
     2581 tgatgcagaa gaaaacactc ggctgggagg cattcaccga gacgctgtac cccgctgacg
     2641 gcggcctgga aggcagaaac gacatggccc tgaagctcgt gggcgggagc catctgatcg
     2701 caaacgccaa gaccacatat agatccaaga aacccgctaa gaacctcaag atgcctggcg
     2761 tctactatgt ggactacaga ctggaaagaa tcaaggaggc caacaacgag acctacgtcg
     2821 agcagcacga ggtggcagtg gccagatacg gtagcggtcc atggaggctg atgcctatta
     2881 cagccgctgc cgaagctatc tgggcctgct ccattgaaga gctgggactg gaaaacgagg
     2941 ccgaaaagcc ttctaatgct ctgctgactc gcgcctggtc cccaggatgg tctaacgctg
     3001 ataaactgct gaatgagttc atcgaaaagc agctgattga ctatgccaaa aactctaaga
     3061 aagtggtggg caacagcacc agcctgctgt ctccatacct gcactttgga gagatcagtg
     3121 tgcggcacgt gttccagtgc gctagaatga agcagatcat ttgggccagg gataaaaaca
     3181 gtgagggcga agagtcagcc gacctgtttc tgaggggaat cggactgagg gagtactccc
     3241 gctatatttg tttcaatttt cccttcactc acgaacagtc actgctgagc catctgcggt
     3301 tctttccttg ggacgctgat gtggacaagt tcaaagcctg gcggcaggga agaacaggct
     3361 atcccctggt ggatgctgga atgagagagc tgtgggccac tggctggatg cacaatagga
     3421 tccgcgtgat tgtgagctcc tttgctgtga agttcctgct gctgccttgg aagtggggca
     3481 tgaaatattt ttgggataca ctgctggatg ccgacctgga gtgtgacatc ctggggtggc
     3541 agtacatctc cggatctatt ccagatggcc atgaactgga taggctggac aaccccgctc
     3601 tgcagggcgc taagtatgac cctgagggag aatacatcag gcagtggctg cctgagctgg
     3661 ctaggctgcc aactgaatgg attcaccatc cctgggacgc tcctctgaca gtgctgaagg
     3721 ccagtggggt ggagctggga actaattacg ctaaacctat cgtggatatt gacaccgcca
     3781 gagagctgct ggctaaggcc atctcccgga ccagagaagc ccagatcatg attggcgctg
     3841 cctctagata agtcgacgga tctttttccc tctgccaaaa attatgggga catcatgaag
     3901 ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt
     3961 tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat
     4021 cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac
     4081 aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta
     4141 ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt
     4201 ttttctttaa catccctaaa attttcctta catgttttac tagccagatt tttcctcctc
     4261 tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct cgacctgcag
     4321 cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4381 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     4441 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4501 gtgccagcgg atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4561 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4621 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     4681 ggcttttttg gaggcctagg cttttgcaaa aagctaactt gtttattgca gcttataatg
     4741 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4801 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cgctgcatta
     4861 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     4921 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4981 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5041 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5101 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5161 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5221 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5281 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5341 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5401 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5461 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5521 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     5581 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     5641 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     5701 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     5761 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     5821 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     5881 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5941 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6001 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6061 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6121 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6181 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6241 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6301 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6361 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6421 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6481 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6541 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     6601 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     6661 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     6721 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6781 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgg
     6841 gtcgatc
//
