LOCUS       sequence-369139-        7631 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid of monSTIM1 (ultra-photosensitive 
            optogenetically-activated STIM1 protein).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7631)
  AUTHORS   Kim S, Kyung T, Chung JH, Kim N, Keum S, Lee J, Park H, Kim HM, Lee 
            S, Shin HS, Do Heo W
  TITLE     Non-invasive optical control of endogenous Ca(2+) channels in awake 
            mice.
  JOURNAL   Nat Commun. 2020 Jan 10;11(1):210. doi: 10.1038/s41467-019-14005-4.
  PUBMED    31924789
REFERENCE   2  (bases 1 to 7631)
  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 
            Commun."; date: "2020-01-10"; volume: "11(1):210. doi"; pages: " 
            10.1038/s41467-019-14005-4"
COMMENT     Sequence Label: pCMV-monSTIM1
FEATURES             Location/Qualifiers
     source          1..7631
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             43..837
                     /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             43..837
                     /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(97..116)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     707..726
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1025..1044)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1069..1116
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1445..2033
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1934..1953
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2208..2511
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2512..2715
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2666..2686
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2744..2753
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2750..2752,2753..2755,2756..3466)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2750..2752,2753..2755,2756..3466)
                     /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(2795..2816)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3056..3075)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3403..3424
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    6566..6687
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6603..6622)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6657..6676
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6694..7149)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6831..6852)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7043..7062
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        7176..7280
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(7282..7631,1..8)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        7282..7473
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(7305..7325)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      7490..7625
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
       61 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
      121 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
      181 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
      241 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
      301 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
      361 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
      421 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
      481 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
      541 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
      601 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
      661 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
      721 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
      781 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
      841 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
      901 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
      961 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     1021 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     1081 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     1141 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     1201 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     1261 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     1321 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     1381 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     1441 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     1501 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     1561 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     1621 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     1681 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     1741 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     1801 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     1861 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     1921 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     1981 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2041 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     2101 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag
     2161 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     2221 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     2281 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     2341 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     2401 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     2461 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     2521 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     2581 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     2641 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     2701 tatataagca gagctggttt agtgaaccgt cagatccgct agcgccacca tggtgagcaa
     2761 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     2821 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     2881 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     2941 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     3001 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     3061 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     3121 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     3181 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     3241 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     3301 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     3361 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     3421 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagccac cggtcatgaa
     3481 gatggacaaa aagaccatcg tctggtttcg gagagatttg agaatagaag ataatcccgc
     3541 gctcgccgcc gcggcccacg agggttccgt cttccccgtt ttcatttggt gtcctgaaga
     3601 agaaggccag ttttatcccg gaagggcctc taggtggtgg atgaagcaaa gtctggccca
     3661 tcttagccag tcactgaaag cactgggcag tgatcttacc ctgatcaaga cacacaatac
     3721 catctctgcc atcctcgact gcatcagggt gaccggcgca acgaaagtcg tgtttaacca
     3781 cctgtacgat ccagttagtc tggtgcgcga ccacactgtg aaggagaagc tggtggaacg
     3841 ggggatcagt gtgcagagct acaacgggga ccttctgtac gagccatggg agatctattg
     3901 cgagaaaggg aaaccgttca cctccttcaa cagttactgg aagaaatgtt tggatatgtc
     3961 aatagagtcc gttatgttgc cccctccctg gagactgatg ccgattactg ctgctgcaga
     4021 ggccatctgg gcctgctcca tcgaggaact cggtctggaa aatgaagcag aaaagccaag
     4081 caatgcactt ctcactagag cctggagccc cggctggtct aatgccgaca agctgcttaa
     4141 cgagttcatc gaaaaacaac tgattgacta cgcgaagaac tccaagaaag tggtaggtaa
     4201 ctcaactagc ttgctctctc catatctcca ttttggcgag atttctgtcc gccatgtatt
     4261 tcagtgcgct cggatgaaac agattatctg ggctcgcgat aaaaacagcg aaggcgccga
     4321 aagcgccgat ctgttcctgc gagggatcgg acttcgggaa tactcccggt atatatgttt
     4381 caactttcca ttcacacacg agcagagtct gttgtcccac ctcaggttct tcccctggga
     4441 cgccgatgtc gacaaattca aggcatggag acagggaagg acagggtacc cactcgtgga
     4501 tgctggcatg agagagctct gggctacagg ctggatgcac aaccgcatcc gggtaatcgt
     4561 gtcctcattt gctgtcaagt ttctgctcct gccttggaaa tggggaatga agtacttttg
     4621 ggataccctt ctcgacgccg acttggagtg tgacattctg ggatggcaat atattagcgg
     4681 gtcaattcct gacggccatg agttggacag gttggacaat ccggccttgc agggagctaa
     4741 gtatgatccc gaaggagagt atattcgaca gtggctcccc gagctggccc gacttcctac
     4801 ggagtggatt caccatcctt gggacgcacc actgacagtg ctcaaggcaa gcggggtgga
     4861 gctgggcacc aattacgcta agcctatagt tgatatagat acagcacgcg agctgctggc
     4921 taaagcgatc tctcgcactc gggaggcgca gattatgatc ggtgctgccg ccagggaccc
     4981 ccccgacctc gacaacgaat tcagtggggg aggcggaggg ggtggttcag gtggtggtgg
     5041 tggttcgggt ggtggcggag ggggaggctc cggactcaga tctaaggagc acatgaagaa
     5101 gatgatgaag gacttggagg ggttacaccg agctgagcag agtctgcatg accttcagga
     5161 aaggctgcac aaggcccagg aggagcaccg cacagtggag gtggagaagg tccatctgga
     5221 aaagaagctg cgcgatgaga tcaaccttgc taagcaggaa gcccagcggc tgaaggagct
     5281 gcgggagggt actgagaatg agcggagccg ccaaaaatat gctgaggagg agttggagca
     5341 ggttcgggag gccttgagga aagcagagaa ggagctagaa tctcacagct catggtatgc
     5401 tccagaggcc cttcagaagt ggctgcagct gacacatgag gtggaggtgc aatattacaa
     5461 catcaagaag caaaatgctg agaagcagct gctggtggcc aaggaggggg ctgagaagat
     5521 aaaaaagaag agaaacacac tctttggcac cttccacgtg gcccacagct cttccctgga
     5581 tgatgtagat cataaaattc taacagctaa gcaagcactg agcgaggtga cagcagcatt
     5641 gcgggagcgc ctgcaccgct ggcaacagat cgagatcctc tgtggcttcc agattgtcaa
     5701 caaccctggc atccactcac tggtggctgc cctcaacata gaccccagct ggatgggcag
     5761 tacacgcccc aaccctgctc acttcatcat gactgacgac gtggatgaca tggatgagga
     5821 gattgtgtct cccttgtcca tgcagtcccc tagcctgcag agcagtgttc ggcagcgcct
     5881 gacggagcca cagcatggcc tgggatctca gagagatttg acccattccg attcggagtc
     5941 ctccctccac atgagtgacc gccagcgtgt ggcccccaaa cctcctcaga tgagccgtgc
     6001 tgcagacgag gctctcaatg ccatgacttc caatggcagc caccggctga tcgagggggt
     6061 ccacccaggg tctctggtgg agaaactgcc tgacagccct gccctggcca agaaggcatt
     6121 actggcgctg aaccatgggc tggacaaggc ccacagcctg atggagctga gcccctcagc
     6181 cccacctggt ggctctccac atttggattc ttcccgttct cacagcccca gctccccaga
     6241 cccagacaca ccatctccag ttggggacag ccgagccctg caagccagcc gaaacacacg
     6301 cattccccac ctggctggca agaaggctgt ggctgaggag gataatggct ctattggcga
     6361 ggaaacagac tccagcccag gccggaagaa gtttcccctc aaaatcttta agaagcctct
     6421 taagaagtag gaattgggat ccaccggatc tagataactg atcataatca gccataccac
     6481 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     6541 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     6601 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     6661 tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa
     6721 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     6781 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     6841 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     6901 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     6961 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     7021 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     7081 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     7141 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     7201 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     7261 aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     7321 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     7381 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     7441 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     7501 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     7561 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     7621 aggcctaggc t
//
