LOCUS       sequence-273675-        7727 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the mechanosensitive channel OSCA4.1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7727)
  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 7727)
  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: OSCA4.1
FEATURES             Location/Qualifiers
     source          1..7727
                     /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(2732..3306,3307..3309,3310..3318)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2899..2916)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3126..3145
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3319..4029
                     /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(3464..3482)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3732..3751
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4151..4272
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4188..4207)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4242..4261
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4279..4734)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4416..4437)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4628..4647
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4761..4865
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4867..5224
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4867..5058
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4890..4910)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5075..5210
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5259..6053
                     /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             5259..6053
                     /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(5313..5332)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5923..5942
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6241..6260)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6285..6332
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6661..7249
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7150..7169
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7424..7727
                     /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 atgaacagaa actttagccc cgccgccatg
      301 cctcctatta gcagcatgac aatcgacaat agcttttccc ctcctccttc cagcggagat
      361 ctgcccgaaa tccccgacgc ttggtacggc aatatccaat acctcctcaa tatcagcgtg
      421 atcggtttac tgtgctgtgt cagcatcttc ctcttcgtga agctgaggag cgaccatcgt
      481 agaatgcccg gccctagcgc cctcttctcc aagttattag ccgtgtggaa agccacatgt
      541 cgtgagatcg ctagacactg cggcgccgat gccgctcagt tcttattaat cgagggcgga
      601 agcttcgtgt tattattcag cattgctgtt ttagccgtga gcgtgatgct gcctttaaat
      661 ctgtatgctg gcacagcttt actgagcgat gagctgtcca agacaatgat tacccacatt
      721 cagaagggca gcgccctcct ctggctgcac ttcgtctttg tggtcatcgt ggtggtgatc
      781 tcccactttg gcatcgctgc tattgaggct cgtctgaagt tcactcgttt tagggatggc
      841 aacggcaata tcagcgaccc caatgccaat agcaccgccg tgtttaccat tatggtgcaa
      901 ggtctgccta agaatctggg aagcgatcgt gtggagttcg aagattgctt tcgtctgaag
      961 taccccggca aagtgtacaa gtttatcgtg cctatggacc tctgcgcttt agatgatctg
     1021 gccaccgagc tcgttcgtgt tcgtgacgag attacttggc tcgtggctaa gatggactct
     1081 cgtttattac ccgatgagta tgagaacgtc ggcgacaatg gtttagtgtt ctgcgtgtgc
     1141 tctttatggg ttcgtgtcaa ggtgctgtgg agccaaatca ccgagaggtt cggcttcacc
     1201 gacgatgaaa agctgaggaa gctgcaagaa ctgagagccg atctcgagtc ccaactggct
     1261 gcctacaaag agggtcgtgc ccaaggcgcc ggcgtcgctt tcgtgatgtt taaggacgtg
     1321 tacaccgcca ataaggccgt gcaagatttt cgtaacgaga ggtctcgtag gactggtaag
     1381 tttttcagcg tgacagagtt acgtctgcag aggaaccagt ggaaggtcga tagggctcct
     1441 ttagccacag acatttactg gaaccattta ggactgacca aggtggcttt aattgtgaga
     1501 agagtgatcg tgaacaccat tttactgctc atcctcgtgt tttttagctc ccctctcgct
     1561 ctgattagcg ctctcgtgtc cgctggcaga attttcaatg ccgaggcttt agattccgct
     1621 cagtactggc tgacttgggt gcagaccagc ggatggattg gctctttaat cttccagttt
     1681 ctgcccaacg tgtttatctt cgtcagcatg tatatcgtca tcccctccgc tttatcctat
     1741 ctgagcaagt tcgaaaggca tctgacagtc agcggcgagc agagagccgc tctgctcaag
     1801 atggtgtgct tctttttagt caacctcatt atcctcaaag ctttagtgga gtcctccctc
     1861 gaatctgctt tactgaagat gagcagatgt tacctcgacg gcgaagactg caaaaggatt
     1921 gaggagtaca tgagccccag ctttctgtct cgtagctgtg tctccgcttt agctttcctc
     1981 atcacctcca ccttcctcgg catcagcttc gatttactgg cccccatccc ttggattaag
     2041 aagaagatcc agaagttcag aaagaatgac atgctgcagc tggtccccga acagaacgag
     2101 gaatacgctt tagaaaacca agaaccctcc tccaatttag agacaccttt actccccgag
     2161 aacatgtttg agagccctag gttcggagac atcgagccca tgtcccaaga tttaagcgag
     2221 taccctatca gcagaaccag ccccatcccc aagcagaagt tcgacttcgc tcagtactac
     2281 gcctttaatt taaccatctt cgccctcacc atgatctaca gctcctttgc ccctttagtc
     2341 gtccccgttg gagccgtgta tttcggatat cgttacatcg tcgataaata caacttttta
     2401 tacgtctatc gtgtgagagg ctttcccgct ggcaatgaag gaaagctgat ggacacagtt
     2461 ttatgtatca tgaggttctg cgtggacctc tatttagtga gcatgttatt attcttctcc
     2521 gtcaagggcg acagcaccaa actgcaagct atcttcactt taggagtttt agtcatgtac
     2581 aaactgctgc ccagcgatac cgatcgttat catcccgcct tactgaggag cattcagacc
     2641 gtggatagca tcatcgacgg acccgtcgat tacgaggcct acagccaccc taacttcgac
     2701 tgggacacct acaacaatcg ttgaggatcc gcccctctcc ctcccccccc cctaacgtta
     2761 ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca
     2821 tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca
     2881 ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg
     2941 aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc
     3001 agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata
     3061 cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag
     3121 tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc
     3181 attgtatggg atctgatctg gggcctcggt acacatgctt tacatgtgtt tagtcgaggt
     3241 taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat
     3301 gataatatgg ccacaaccat ggtgagcaag ggcgaggagg ataacatggc catcatcaag
     3361 gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc
     3421 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
     3481 aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc
     3541 aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag
     3601 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag
     3661 gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc
     3721 ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg
     3781 atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac
     3841 ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg
     3901 cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc
     3961 atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg
     4021 tacaagtaag cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt
     4081 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     4141 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4201 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4261 caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     4321 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     4381 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     4441 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     4501 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     4561 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     4621 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     4681 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     4741 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4801 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4861 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4921 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4981 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     5041 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     5101 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     5161 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     5221 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     5281 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     5341 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     5401 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     5461 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     5521 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     5581 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     5641 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5701 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5761 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     5821 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5881 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5941 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     6001 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     6061 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     6121 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     6181 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     6241 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     6301 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     6361 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     6421 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     6481 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     6541 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6601 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6661 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6721 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6781 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     6841 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6901 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6961 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     7021 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     7081 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     7141 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     7201 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     7261 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     7321 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
     7381 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     7441 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     7501 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     7561 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     7621 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     7681 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatg
//
