LOCUS       sequence-273671-        7385 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the mechanosensitive channel OSCA2.3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7385)
  AUTHORS   Murthy SE, Dubin AE, Whitwam T, Jojoa-Cruz S, Cahalan SM, Mousavi 
            SAR, Ward AB, Patapoutian A
  TITLE     OSCA/TMEM63 are an Evolutionarily Conserved Family of Mechanically 
            Activated Ion Channels.
  JOURNAL   Elife. 2018 Nov 1;7. pii: 41844. doi: 10.7554/eLife.41844.
  PUBMED    30382938
REFERENCE   2  (bases 1 to 7385)
  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: "Elife."; 
            date: "2018-11-1"; volume: "7. pii: 41844. doi"; pages: " 
            10.7554/eLife.41844"
COMMENT     Sequence Label: OSCA2.3
FEATURES             Location/Qualifiers
     source          1..7385
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    join(2694..3268,3269..3271,3272..3280)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2861..2878)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3088..3107
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3281..3991
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3426..3444)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3694..3713
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4113..4234
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4150..4169)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4204..4223
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4241..4696)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4378..4399)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4590..4609
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4723..4827
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4829..5186
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4829..5020
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4852..4872)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5037..5172
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5221..6015
                     /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             5221..6015
                     /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(5275..5294)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5885..5904
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6203..6222)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6247..6294
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6623..7211
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7112..7131
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcga attcatgtta ttaagcgcct tattaacaag
      601 cgtgggaatt aatttaggtt tatgctttct gttcttcaca ctgtacagca ttttacgtaa
      661 acagcccagc aacgtgacag tgtatggccc tagactggtg aagaaggacg gcaagagcca
      721 gcaaagcaac gagttcaatt tagagagatt attacctacc gccggctggg tgaagagagc
      781 cctcgagcct accaacgacg aaattctgtc caatttaggc ctcgatgctt tagtcttcat
      841 cagagtcttc gtgttctcca ttcgtgtgtt tagcttcgcc agcgtcgtgg gcatcttcat
      901 tctgctgccc gtgaactaca tgggcaccga gtttgaggag tttttcgatc tccctaagaa
      961 gtccatggat aatttctcca ttagcaatgt caacgatgga agcaataagc tgtggattca
     1021 tttctgcgcc atctacattt tcaccgccgt cgtctgcagc ttattatatt acgaacataa
     1081 gtacatttta accaagagga tcgcccatct ctactccagc aagccccagc ctcaagagtt
     1141 tacagtgctg gtgagcggcg tgcctctggt ctccggcaat agcatctccg agaccgtgga
     1201 gaacttcttc agagaatacc atagctcctc ctatttatcc cacatcgtcg tgcacagaac
     1261 cgacaaactc aaggtgctga tgaacgacgc cgagaagctg tacaagaagc tgactcgtgt
     1321 gaagagcggc tccatctctc gtcaaaagag cagatggggc ggatttctgg gcatgttcgg
     1381 caacaacgtg gatgtggtcg accactatca gaagaaactc gacaagctcg aggacgatat
     1441 gaggctcaag cagtctttac tggctggcga agaggtgccc gctgccttcg tgtccttcag
     1501 aacaagacac ggcgctgcca tcgccaccaa catccaacaa ggtattgacc ccacacagtg
     1561 gctgaccgag gctgctcccg aacccgaaga tgtgcattgg cctttcttca cagcctcctt
     1621 cgtgaggagg tggatcagca atgtggtggt gctcgtggcc ttcgtcgctc tgctgatcct
     1681 ctacatcgtg cccgttgtgc tggtgcaagg tttagccaat ttacatcagc tcgagacttg
     1741 gttccccttt ctgaagggca tcctcaacat gaagatcgtg tcccaagtta tcaccggcta
     1801 tttaccttct ttaatcttcc agctgttcct cctcatcgtg cctcccatta tgctgctcct
     1861 ctccagcatg caaggtttca ttagccactc ccagatcgag aagtccgctt gcatcaaatt
     1921 attaatcttc accgtgtgga actctttctt cgccaatgtt ttatccggct ccgctctgta
     1981 cagagtgaat gtgtttttag agcccaagac cattcctaga gttttagccg ctgctgtgcc
     2041 cgctcaagcc tccttcttcg tgagctatgt ggtgacaagc ggatggaccg gtttaagcag
     2101 cgagatttta aggctggtcc ccctcctctg gagctttatt accaaactct tcggcaagga
     2161 ggatgacaag gagttcgagg tcccctccac ccccttctgc caagaaatcc cccgtatttt
     2221 attcttcggt ttactgggaa tcacctactt tttcctctcc cctttaattc tccccttctt
     2281 attagtgtac tactgcctcg gatacattat ctatcgtaac cagctgctga acgtgtacgc
     2341 cgctaagtac gagaccggcg gaaaattctg gcccatcgtg cacagctata ccatcttctc
     2401 cctcgtgctg atgcatatca ttgccgtcgg tttatttggt ttaaaggagc tccccgtcgc
     2461 ctcctcttta accatccctc tgcccgtcct caccgtgctc ttcagcatct actgccagag
     2521 gagattcctc cctaacttca aatcctaccc tacccagtgc ctcgtgaaca aggacaaagc
     2581 cgacgagagg gagcagaata tgagcgagtt ctacagcgag ctggtcgtgg cctataggga
     2641 ccccgcttta agcgccagcc aagattctcg tgacatctcc ccctgaggat ccgcccctct
     2701 ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
     2761 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
     2821 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa
     2881 ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg
     2941 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc
     3001 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg
     3061 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg
     3121 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtacacatgc
     3181 tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg
     3241 gttttccttt gaaaaacacg atgataatat ggccacaacc atggtgagca agggcgagga
     3301 ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt
     3361 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
     3421 gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc
     3481 ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta
     3541 cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
     3601 cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt
     3661 gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg
     3721 ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa
     3781 gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa
     3841 ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac
     3901 ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc
     3961 caccggcggc atggacgagc tgtacaagta agcggccgcg actctagatc ataatcagcc
     4021 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     4081 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     4141 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4201 gttgtggttt gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt
     4261 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     4321 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     4381 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     4441 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     4501 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     4561 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     4621 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     4681 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4741 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4801 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     4861 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4921 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4981 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     5041 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5101 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     5161 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     5221 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     5281 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     5341 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     5401 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     5461 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     5521 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     5581 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     5641 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     5701 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     5761 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     5821 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     5881 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     5941 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     6001 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     6061 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     6121 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     6181 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     6241 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     6301 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     6361 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     6421 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     6481 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     6541 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6601 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6661 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6721 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6781 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6841 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6901 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6961 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     7021 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     7081 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     7141 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     7201 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     7261 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca
     7321 tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     7381 ttccg
//
