LOCUS       sequence-273753-        7568 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the mechanosensitive channel Drosophila melanogaster 
            Tmem63 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7568)
  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 7568)
  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: DmTmem63
FEATURES             Location/Qualifiers
     source          1..7568
                     /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(2573..3147,3148..3150,3151..3159)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2740..2757)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2967..2986
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3160..3870
                     /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(3305..3323)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3573..3592
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3992..4113
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4029..4048)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4083..4102
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4120..4575)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4257..4278)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4469..4488
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4602..4706
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4708..5065
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4708..4899
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4731..4751)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4916..5051
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5100..5894
                     /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             5100..5894
                     /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(5154..5173)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5764..5783
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6082..6101)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6126..6173
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6502..7090
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6991..7010
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7265..7568
                     /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 tgatgtccga aaattctaac
      301 atcagctact gccaggctgt cgccaatagg accagctttt tcaccaccgc ctacaatggc
      361 atccccgaga ccctcatcct gaacaccatc ttttgggtcc tgctcatcct cctcttcaca
      421 gtgctgaggc atcaagccgg agacttcggc agactggccc tggtgaatag caacggcagc
      481 aagaagaggt ggaccgagat cttctatagc agagctgcca gcgtggttga tcctcagccc
      541 agcacaagca cccaggtgag ccccaggccc agcacaacaa gccagggcgt gcctgatagc
      601 acccccctgt cccccatcca acccgaagag ggcctcttcg gctggatcag agtgacactg
      661 aagctcagga aggagaccat cctgctgcat acaggacccg acgccgtgca ctatctcagc
      721 ttccaacagc acctgatggc tgtcatggcc ctggtcacca tcgtgagcct ggtgatcatc
      781 ctgcccgtga acttcctgaa cggccctaag gaaaacccct atgacgtgaa cgccttcggc
      841 aggaccacaa tggccaacct gtcccctgac agcccttggc tgtgggtcca cacaatcatc
      901 accattctgt atatccccct cgtggtcctg atcatgagga gggccagcgg aaggaacgct
      961 ttcaagaaag ccgccaccag gaccatcatg atctccaata tcagctccag cgacaggaac
     1021 aagacagtcg tgaggaacta catgcaggag ctgttccccg atgtgacaat cgagaacgtg
     1081 agcatcgcct acaacattag caggctctac gtgagaaatg ccgaatttga gagagtgcac
     1141 gaggctaggc tgtactgcga acaccacaga aacagagaca ccctgatggt gaagcccgaa
     1201 atgtgctcct gcaagaagga gaacgcctac gaatactatc aaagagagga gaggaagctg
     1261 tccggcgacg tggccagact gagagcctcc accatgaacg agcccctcga tatcgctttc
     1321 attaccgtgt ccaccgtcca agaagcccag aacatcgtga cccacttcac acccggaacc
     1381 tataggcagt ggcatctcgt cttcgcccct tcccccgacg atctcttctg ggagaatctc
     1441 aacgtcaaca agagccactg gtatctgaaa ttcttctgtg tcaacgccgt gctgttcctg
     1501 gtcctgttct tcctgagcac acccgccatg gtcgtgagcc tgctgaatag cagaccctgg
     1561 gtgaaggaga ccgagggcaa gatcagcccc ctggtggccg attttctgcc caccctgatg
     1621 ctgtggaccc tgagcgctct catgcctgtc atcgtcgcca tcagcgataa gtggatgaga
     1681 cactacacca ggagcaaaca gaactacagc attatgacca aatgcttcgg ctacctcctg
     1741 atgatgatca tgatcctgcc ctccctggga ctcaccagcg ctcaagctct cttagaatgg
     1801 ggcttcctga atcagaccga cagatggcag tgcattttcc tccctgacag gggctccttc
     1861 tacgtgaact acatcattac cgctgctttt atcggcacag ctctggagct gctgagattc
     1921 cctgagctga tcgtctatat ctggtccctc ctgaaggcca agagcaaagc cgagacccct
     1981 tacatcagga aggccatcct catcgagttc cccttcggaa cccactacgc ctggacaacc
     2041 ctggtgttca ccatcgctat cgtctattcc gtggcctgtc ctctgattat gcctttcgcc
     2101 atgatctaca tctgcctcaa gcacttcgtc gacagacaca atctgtactt cgcttatggc
     2161 cccagcaaca tgattagcag gaagggcgga aagatccata gcaccgccgt caccatgacc
     2221 aagttctccg tcgtcattct ggtgctcgtc atggccatga tttgcttctt caggggagac
     2281 aacggcatgg ctaggttcct gctgctcatc atcagcctgg ccatcaccct gacactgttc
     2341 accttcatga gccccatcaa gagatgcgcc gccaccagga ggccttccat cgtggaaatc
     2401 gccggccccg cccctatcta tgtgcccgat gtgctgaagg ataacggcat caggacatcc
     2461 agcgtcagac cctccgccgc tgctttcggc ggatatggct ccgactccgt gtccgaatat
     2521 gattccagcc aatacagcgt gaactccgtg gaggcctgac cgcggggatc cgcccctctc
     2581 cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
     2641 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
     2701 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
     2761 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt
     2821 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
     2881 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
     2941 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
     3001 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tacacatgct
     3061 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     3121 ttttcctttg aaaaacacga tgataatatg gccacaacca tggtgagcaa gggcgaggag
     3181 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
     3241 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     3301 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
     3361 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
     3421 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     3481 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
     3541 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
     3601 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
     3661 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
     3721 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc
     3781 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
     3841 accggcggca tggacgagct gtacaagtaa gcggccgcga ctctagatca taatcagcca
     3901 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     3961 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     4021 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4081 ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt
     4141 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     4201 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     4261 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     4321 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     4381 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     4441 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     4501 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     4561 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     4621 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     4681 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     4741 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4801 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4861 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     4921 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     4981 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     5041 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     5101 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     5161 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     5221 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     5281 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     5341 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     5401 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     5461 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     5521 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     5581 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     5641 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     5701 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     5761 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     5821 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     5881 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     5941 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     6001 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     6061 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     6121 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     6181 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     6241 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     6301 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     6361 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6421 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     6481 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6541 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6601 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6661 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6721 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6781 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6841 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6901 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6961 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     7021 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     7081 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     7141 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
     7201 gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     7261 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     7321 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     7381 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     7441 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     7501 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     7561 ttaccatg
//
