LOCUS       sequence-497312-        6944 bp    DNA     circular SYN 14-JAN-2026
DEFINITION  An expression construct encoding a CRY2-fused FLAG-tagged STIM1 
            fragment (a.a.238–685)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6944)
  AUTHORS   Kook YH, Lee H, Lee J, Jeong Y, Rho J, Heo WD, Lee S
  TITLE     AAV-compatible optogenetic tools for activating endogenous calcium 
            channels in vivo.
  JOURNAL   Mol Brain. 2023 Oct 17;16(1):73. doi: 10.1186/s13041-023-01061-7.
  PUBMED    37848907
REFERENCE   2  (bases 1 to 6944)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 14, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol 
            Brain."; date: "2023-10-17"; volume: "16(1):73. doi"; pages: " 
            10.1186/s13041-023-01061-7"
COMMENT     Sequence Label: pCMV-FLAG-CRY2-STIM1(238-685)
FEATURES             Location/Qualifiers
     source          1..6944
                     /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"
     regulatory      597..606
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             609..632
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3732..3853
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3769..3788)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3823..3842
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3860..4315)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3997..4018)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4209..4228
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4342..4446
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4448..4805
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4448..4639
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4471..4491)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4656..4791
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4840..5634
                     /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             4840..5634
                     /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(4894..4913)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5504..5523
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5822..5841)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5866..5913
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6242..6830
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6731..6750
                     /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 gctagcgcca
      601 ccatgggaga ctacaaggat gacgacgata agccaccggt catgaagatg gacaaaaaga
      661 ccatcgtctg gtttcggaga gatttgagaa tagaagataa tcccgcgctc gccgccgcgg
      721 cccacgaggg ttccgtcttc cccgttttca tttggtgtcc tgaagaagaa ggccagtttt
      781 atcccggaag ggcctctagg tggtggatga agcaaagtct ggcccatctt agccagtcac
      841 tgaaagcact gggcagtgat cttaccctga tcaagacaca caataccatc tctgccatcc
      901 tcgactgcat cagggtgacc ggcgcaacga aagtcgtgtt taaccacctg tacgatccag
      961 ttagtctggt gcgcgaccac actgtgaagg agaagctggt ggaacggggg atcagtgtgc
     1021 agagctacaa cggggacctt ctgtacgagc catgggagat ctattgcgag aaagggaaac
     1081 cgttcacctc cttcaacagt tactggaaga aatgtttgga tatgtcaata gagtccgtta
     1141 tgttgccccc tccctggaga ctgatgccga ttactgctgc tgcagaggcc atctgggcct
     1201 gctccatcga ggaactcggt ctggaaaatg aagcagaaaa gccaagcaat gcacttctca
     1261 ctagagcctg gagccccggc tggtctaatg ccgacaagct gcttaacgag ttcatcgaaa
     1321 aacaactgat tgactacgcg aagaactcca agaaagtggt aggtaactca actagcttgc
     1381 tctctccata tctccatttt ggcgagattt ctgtccgcca tgtatttcag tgcgctcgga
     1441 tgaaacagat tatctgggct cgcgataaaa acagcgaagg cgccgaaagc gccgatctgt
     1501 tcctgcgagg gatcggactt cgggaatact cccggtatat atgtttcaac tttccattca
     1561 cacacgagca gagtctgttg tcccacctca ggttcttccc ctgggacgcc gatgtcgaca
     1621 aattcaaggc atggagacag ggaaggacag ggtacccact cgtggatgct ggcatgagag
     1681 agctctgggc tacaggctgg atgcacaacc gcatccgggt aatcgtgtcc tcatttgctg
     1741 tcaagtttct gctcctgcct tggaaatggg gaatgaagta cttttgggat acccttctcg
     1801 acgccgactt ggagtgtgac attctgggat ggcaatatat tagcgggtca attcctgacg
     1861 gccatgagtt ggacaggttg gacaatccgg ccttgcaggg agctaagtat gatcccgaag
     1921 gagagtatat tcgacagtgg ctccccgagc tggcccgact tcctacggag tggattcacc
     1981 atccttggga cgcaccactg acagtgctca aggcaagcgg ggtggagctg ggcaccaatt
     2041 acgctaagcc tatagttgat atagatacag cacgcgagct gctggctaaa gcgatctctc
     2101 gcactcggga ggcgcagatt atgatcggtg ctgccgccag ggaccccccc gacctcgaca
     2161 acgaattcag tgggggaggc ggagggggtg gttcaggtgg tggtggtggt tcgggtggtg
     2221 gcggaggggg aggctccgga ctcagatcta aggagcacat gaagaagatg atgaaggact
     2281 tggaggggtt acaccgagct gagcagagtc tgcatgacct tcaggaaagg ctgcacaagg
     2341 cccaggagga gcaccgcaca gtggaggtgg agaaggtcca tctggaaaag aagctgcgcg
     2401 atgagatcaa ccttgctaag caggaagccc agcggctgaa ggagctgcgg gagggtactg
     2461 agaatgagcg gagccgccaa aaatatgctg aggaggagtt ggagcaggtt cgggaggcct
     2521 tgaggaaagc agagaaggag ctagaatctc acagctcatg gtatgctcca gaggcccttc
     2581 agaagtggct gcagctgaca catgaggtgg aggtgcaata ttacaacatc aagaagcaaa
     2641 atgctgagaa gcagctgctg gtggccaagg agggggctga gaagataaaa aagaagagaa
     2701 acacactctt tggcaccttc cacgtggccc acagctcttc cctggatgat gtagatcata
     2761 aaattctaac agctaagcaa gcactgagcg aggtgacagc agcattgcgg gagcgcctgc
     2821 accgctggca acagatcgag atcctctgtg gcttccagat tgtcaacaac cctggcatcc
     2881 actcactggt ggctgccctc aacatagacc ccagctggat gggcagtaca cgccccaacc
     2941 ctgctcactt catcatgact gacgacgtgg atgacatgga tgaggagatt gtgtctccct
     3001 tgtccatgca gtcccctagc ctgcagagca gtgttcggca gcgcctgacg gagccacagc
     3061 atggcctggg atctcagaga gatttgaccc attccgattc ggagtcctcc ctccacatga
     3121 gtgaccgcca gcgtgtggcc cccaaacctc ctcagatgag ccgtgctgca gacgaggctc
     3181 tcaatgccat gacttccaat ggcagccacc ggctgatcga gggggtccac ccagggtctc
     3241 tggtggagaa actgcctgac agccctgccc tggccaagaa ggcattactg gcgctgaacc
     3301 atgggctgga caaggcccac agcctgatgg agctgagccc ctcagcccca cctggtggct
     3361 ctccacattt ggattcttcc cgttctcaca gccccagctc cccagaccca gacacaccat
     3421 ctccagttgg ggacagccga gccctgcaag ccagccgaaa cacacgcatt ccccacctgg
     3481 ctggcaagaa ggctgtggct gaggaggata atggctctat tggcgaggaa acagactcca
     3541 gcccaggccg gaagaagttt cccctcaaaa tctttaagaa gcctcttaag aagtaggaat
     3601 tgggatccac cggatctaga taactgatca taatcagcca taccacattt gtagaggttt
     3661 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     3721 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     3781 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     3841 tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     3901 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     3961 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     4021 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     4081 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     4141 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     4201 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     4261 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     4321 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     4381 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4441 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4501 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4561 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4621 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4681 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4741 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4801 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     4861 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     4921 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     4981 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     5041 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     5101 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     5161 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     5221 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     5281 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     5341 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     5401 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     5461 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     5521 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     5581 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     5641 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     5701 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     5761 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
     5821 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     5881 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     5941 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     6001 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     6061 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
     6121 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6181 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6241 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6301 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6361 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     6421 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6481 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6541 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6601 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6661 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6721 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6781 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6841 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     6901 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
