LOCUS       sequence-273740-        7601 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the mechanosensitive channel OSCA1.2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7601)
  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 7601)
  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: OSCA1.2
FEATURES             Location/Qualifiers
     source          1..7601
                     /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(2606..3180,3181..3183,3184..3192)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2773..2790)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3000..3019
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3193..3903
                     /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(3338..3356)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3606..3625
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4025..4146
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4062..4081)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4116..4135
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4153..4608)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4290..4311)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4502..4521
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4635..4739
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4741..5098
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4741..4932
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4764..4784)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4949..5084
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5133..5927
                     /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             5133..5927
                     /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(5187..5206)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5797..5816
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6115..6134)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6159..6206
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6535..7123
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7024..7043
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7298..7601
                     /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 ctcgagatgg ctaccctgca agatatcggc
      301 gtgagcgctg gaatcaacat cctctccgcc tttgtgttct tcatcatctt cgctgtgctg
      361 agactgcagc cctttaacga cagggtctac ttcagcaagt ggtacctgaa gggactcagg
      421 tccagccctg ctaggggagg agccttcgct cagagattcg tgaacctcga cttcaggagc
      481 tacatgaagt tcctgaactg gatgcctgag gctctcaaaa tgcccgagcc cgagctgatt
      541 gatcacgccg gcctggatag cgtcgtctat ctgaggatct actggctcgg cctgaagatc
      601 tttacaccta tcgccgtgct ggcctgggct gtcctggtgc ccgtgaattg gaccaacaac
      661 acactggaga tggccaaaca gctgaggaat gtgacctcct ccgacatcga caaactgagc
      721 gtctccaaca tccccgagta tagcatgagg ttctggaccc acatcgtcat ggcctacgcc
      781 tttaccatct ggacctgcta cgtcctgatg aaggagtacg agaccatcgc caacatgagg
      841 ctgcagttcg tggccagcga agctagaagg cccgaccagt ttaccgtgct ggtgaggaac
      901 gtgccccctg acgctgacga atccgtgagc gagctcgtcg agcatttctt cctggtgaac
      961 catcccgacc actacctgac ccaccaagtg gtgtgcaacg ccaacaaact cgccgacctg
     1021 gtcaaaaaaa aaaagaagct ccagaactgg ctggactact accagctcaa gtacgccagg
     1081 aacaatagcc agaggattat ggtcaagctc ggctttctgg gcctctgggg ccagaaggtc
     1141 gatgccattg agcactacat cgccgagatt gacaaaatct ccaaggagat ctccaaagag
     1201 agggaggaag tggtgaacga ccccaaagcc atcatgcctg ccgccttcgt gagctttaag
     1261 accaggtggg ctgccgccgt gtgcgctcag acccaacaga ccagaaaccc cacccagtgg
     1321 ctcaccgaat gggctcctga acccagggac gtgttctggt ccaatctggc catcccctat
     1381 gtgagcctga ccgtgaggag gctgattatg cacgtggcct tcttcttcct gacctttttc
     1441 tttatcgtcc ctatcgcctt cgtccagtcc ctggctacca tcgaaggaat tgtcaaagcc
     1501 gcccccttcc tcaagttcat tgtcgacgac aagttcatga agagcgtgat ccagggattc
     1561 ctccctggca ttgctctgaa gctgttcctg gcttttctcc cctccatcct gatgatcatg
     1621 agcaagttcg agggcttcac ctccatctcc agcttagaaa ggagggccgc cttcagatac
     1681 tatattttta acctggtgaa tgtctttctg gcctccgtga ttgctggagc cgccttcgag
     1741 cagctcaaca gcttcctcaa ccagtccgct aaccagattc ctaagaccat cggcgtggct
     1801 atccccatga aagccacatt ctttatcaca tacattatgg tggacggatg ggccggcgtg
     1861 gctggagaaa tcctcatgct gaagcctctg attatgtttc atctgaaaaa tgcttttctg
     1921 gtgaagacag acaaggatag ggaggaagcc atggaccctg gttctattgg ctttaacaca
     1981 ggcgagccca ggattcagct gtacttcctg ctgggactcg tctacgcccc cgtgacaccc
     2041 atgctgctgc cctttatcct cgtgttcttc gctctggcct acattgtcta caggcaccag
     2101 atcatcaacg tgtacaatca ggagtacgaa tccgccgctg ccttctggcc cgacgtgcat
     2161 ggcagagtga ttgctgccct ggtgatcagc cagctgctcc tcatgggact gctcggaacc
     2221 aagcatgccg ctctggccgc cccctttctg attgctctgc ccgtcctgac catcggcttt
     2281 caccatttct gcaagggcag gtacgagcct gctttcatca ggtaccccct gcaggaagcc
     2341 atgatgaagg acaccttaga aacagccaga gagcctaacc tcaacctcaa gggctacctg
     2401 cagaatgcct acgtgcaccc cgtgttcaag ggcgacgagg atgactatga catcgatgac
     2461 aaactgggca agtttgagga cgaggctatc atcgtcccca ccaaaaggca atccaggaga
     2521 aacacccctg cccccagcat tatctccggc gatgatagcc ctagcctgcc cttttccggc
     2581 aagctcgtgt gaccgcgggg atccgcccct ctccctcccc cccccctaac gttactggcc
     2641 gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc
     2701 cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta
     2761 ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag
     2821 ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga
     2881 accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg
     2941 caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat
     3001 ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta
     3061 tgggatctga tctggggcct cggtacacat gctttacatg tgtttagtcg aggttaaaaa
     3121 aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat
     3181 atggccacaa ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
     3241 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
     3301 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
     3361 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
     3421 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
     3481 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
     3541 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     3601 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     3661 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     3721 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     3781 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     3841 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     3901 taagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
     3961 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     4021 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4081 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4141 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     4201 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     4261 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     4321 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     4381 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     4441 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     4501 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     4561 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     4621 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4681 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     4741 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     4801 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     4861 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4921 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4981 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     5041 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     5101 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     5161 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     5221 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     5281 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     5341 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5401 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5461 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5521 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     5581 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5641 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5701 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5761 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5821 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5881 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5941 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     6001 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     6061 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     6121 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     6181 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     6241 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     6301 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     6361 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     6421 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6481 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6541 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6601 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6661 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     6721 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6781 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6841 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6901 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6961 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     7021 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     7081 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     7141 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     7201 ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta
     7261 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     7321 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     7381 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     7441 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     7501 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     7561 cttggcagta catctacgta ttagtcatcg ctattaccat g
//
