LOCUS       sequence-273738-        7694 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the mechanosensitive channel OSCA1.8 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7694)
  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 7694)
  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: OSCA1.8
FEATURES             Location/Qualifiers
     source          1..7694
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    join(2699..3273,3274..3276,3277..3285)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2866..2883)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3093..3112
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3286..3996
                     /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(3431..3449)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3699..3718
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4118..4239
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4155..4174)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4209..4228
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4246..4701)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4383..4404)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4595..4614
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4728..4832
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4834..5191
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4834..5025
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4857..4877)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5042..5177
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5226..6020
                     /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             5226..6020
                     /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(5280..5299)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5890..5909
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6208..6227)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6252..6299
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6628..7216
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7117..7136
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7391..7694
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 actcagatct cgagctcaag cttcgaattc atggccactt tacaagacat tggcgtctcc
      301 gctctgatca atctcttcgg cgccttttta tttttaatcg ctttcgccgt gctgcgtatt
      361 caacccatca acgatcgtgt gtacttcccc aagtggtatt taactggtga gagaaacagc
      421 cctaggagaa gcgacagaac tttagtcggc aaattcgtca atttaaatta taaaacctac
      481 ttcacctttt taaattggat gccccaagct atgaaaatga gcgaatccga gattatcaga
      541 cacgccggac tcgacagcgc catcttttta aggatttaca ctttaggact gaagatcttt
      601 gcccccgtta tggtgctggc tctcgtggtc ctcgtccccg ttaatgtgag ctccggcact
      661 ttattctttc tcaagaagga actggtggtg agcaacatcg ataagctctc catcagcaac
      721 gtgcagccta agagctccaa gttcttcttc cacatcgccg tggagtatat ttttaccttc
      781 tgggcttgtt ttatgctcta tcgtgagtac aacaatgtgg ccatcatgag gctgcagtat
      841 ttagcctccc agaggaggag gcccgaacag ttcacagtgg tggtgagaaa tgtccccgac
      901 atgcccggcc attccgtgcc cgataccgtg gatcaatttt tcaaaaccaa tcaccccgag
      961 cactatttat gccatcaagc tgtgtacaat gccaatacct acgccaagct ggtcaagcag
     1021 agggccaagc tgcagaggtg gttcgactac tatgttttaa agcaccagag gaacccccac
     1081 aagcagccta catgtaggac cggatttctc ggcctctggg gcaagagggt ggactccatc
     1141 gagtattata aacaacagat caaggagttt gaccataaca tgagcctcga gaggcagaag
     1201 gttttaaagg actccaagct gatgctgccc gttgcctttg tgagctttga ctctcgttgg
     1261 ggagctgccg tgtgcgccca gacccaacag agcaagaatc ccacactgtg gctgaccagc
     1321 tccgcccccg aacctagaga catctactgg cagaatttag ccatcccctt catctccctc
     1381 acaattcgta agctggtcat tggcgtgagc gtgttcgccc tcgtgttttt ctacatgatt
     1441 cccatcgcct tcgtgcagag cctcgccaat ctggagggcc tcgatagagt ggctcccttc
     1501 ctcagacccg tcacaaggct cgacttcatc aagtcctttt tacaaggttt tctgcccgga
     1561 ctggccctca aaatcttcct ctggatttta cctacagtct tattaatcat gagcaagatt
     1621 gagggctata ttgctttaag cacactggaa aggagggccg ctgccaagta ctattacttc
     1681 atgctggtca acgtcttttt aggaagcatt atcgctggca ccgccttcga acagctgcac
     1741 tcctttctcc accagtcccc tagccagatt cctcgtacaa tcggcgttag cattcccatg
     1801 aaagccacct tcttcattac ctacatcatg gtggatggat gggctggcat cgccggagag
     1861 attttaaggc tgaaacctct ggtcatcttc catctgaaaa acatgttcat cgtgaagacc
     1921 gaagaggaca gagtgagagc catggacccc ggttttgtcg acttcaagga gacaatcccc
     1981 tctttacagc tgtactttct gctgggcatt gtctataccg ccgtgacccc tattttactg
     2041 ccctttattc tgattttctt cgctttcgcc tatctcgtgt atcgtcatca gatcatcaat
     2101 gtgtataacc agcagtacga gtcttgtgga gccttctggc ctcacgtgca cggtcgtatc
     2161 atcgcttctt tactcattag ccaactgtta ttaatgggtt tactggccag caagaaagct
     2221 gccgattcca cccctttact gatcatcctc cccattttaa ctttatcctt ccataagtac
     2281 tgcaaacaca gatttgagcc cgcctttcgt cagtaccctt tagaggaggc catggccaaa
     2341 gacaagctgg aaaaggagac cgagcccgaa ctgaacatga aggctgattt agccgacgcc
     2401 tacctccacc ctatcttcca cagcttcgag aaagaagtgg agctgagcag cagcagctcc
     2461 agcgaaaagg aaacccatca agaagagacc cccgaagtga gagtcgacaa gcatgagacc
     2521 cagtcctcct cccccgttac cgagctgggc acaagcagcc accaccacca cgtgtacaac
     2581 agcacatccc cctcctccca ctacgccagc gcttatgagc agagcagctc ccaatacgaa
     2641 taccactaca atacccacca gtacgaggaa catgagtaca gatataattg aggatccgcc
     2701 cctctccctc ccccccccnt aacgttactg gccgaagccg cttggaataa ggccggtgtg
     2761 cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga
     2821 aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa
     2881 tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa
     2941 caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct
     3001 gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg
     3061 ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg
     3121 ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtaca
     3181 catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg
     3241 acgtggtttt cctttgaaaa acacgatgat aatatggcca caaccatggt gagcaagggc
     3301 gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc
     3361 tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     3421 acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc
     3481 ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc
     3541 gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag
     3601 gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac
     3661 aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc
     3721 atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag
     3781 atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc
     3841 tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac
     3901 atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc
     3961 cactccaccg gcggcatgga cgagctgtac aagtaagcgg ccgcgactct agatcataat
     4021 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     4081 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     4141 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4201 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta
     4261 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     4321 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     4381 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     4441 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     4501 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     4561 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     4621 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     4681 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     4741 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     4801 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     4861 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     4921 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     4981 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     5041 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     5101 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     5161 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     5221 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     5281 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     5341 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     5401 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     5461 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     5521 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     5581 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     5641 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     5701 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     5761 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     5821 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     5881 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     5941 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     6001 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     6061 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     6121 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     6181 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     6241 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     6301 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     6361 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     6421 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     6481 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     6541 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     6601 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     6661 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6721 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6781 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6841 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6901 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6961 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     7021 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     7081 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     7141 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     7201 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     7261 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     7321 cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     7381 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     7441 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     7501 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     7561 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     7621 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     7681 tcgctattac catg
//
