LOCUS       sequence-273676-        7460 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses the mechanosensitive channel OSCA3.1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7460)
  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 7460)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: OSCA3.1
FEATURES             Location/Qualifiers
     source          1..7460
                     /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(2465..3039,3040..3042,3043..3051)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2632..2649)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2859..2878
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3052..3762
                     /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(3197..3215)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3465..3484
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3884..4005
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3921..3940)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3975..3994
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4012..4467)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4149..4170)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4361..4380
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4494..4598
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4600..4957
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4600..4791
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4623..4643)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4808..4943
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4992..5786
                     /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             4992..5786
                     /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(5046..5065)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5656..5675
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5974..5993)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6018..6065
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6394..6982
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6883..6902
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7157..7460
                     /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 agtttggcag cttcctggtg
      301 agcctgggca cctccttcgt gatcttcgtg atcctgatgc tgctgttcac atggctgagc
      361 aggaaaagcg gcaacgctcc catctactac cccaaccgta ttttaaaggg cctcgaacct
      421 tgggagggca catccctcac aaggaacccc ttcgcctgga tgagggaggc cctgacctcc
      481 agcgagcagg acgtggtgaa cctgtccgga gtggacaccg ccgtccactt tgtgttcctc
      541 tccaccgtgc tgggcatttt cgcctgttcc tccctcctgc tgctgcccac actgctccct
      601 ctcgctgcca cagacaataa cattaagaac accaaaaacg ccacagacac cacaagcaag
      661 ggcacattct cccagctgga caacctgtcc atggccaaca tcaccaagaa gagctccaga
      721 ctctgggctt tcctgggcgc cgtgtactgg atctccctgg tgacctactt cttcctgtgg
      781 aaggcttaca aacatgtgtc ctccctcagg gcccaggccc tcatgtccgc tgacgtgaaa
      841 cctgaacaat tcgccatcct ggtgagagac atgcctgccc ctcccgacgg acagacccag
      901 aaggagttta ttgacagcta cttcagggag atctaccccg agaccttcta cagaagcctg
      961 gtggccaccg aaaacagcaa ggtcaacaaa atctgggaga agctcgaagg ctataagaag
     1021 aagctcgcta gagccgaggc catcctggct gctacaaata acaggcccac caacaaaacc
     1081 ggcttctgtg gcctggtcgg aaagcaggtg gactccatcg agtactacac agagctcatt
     1141 aacgagtccg tcgccaaatt agaaaccgag cagaaagccg tgctggccga gaagcagcag
     1201 acagctgccg tcgtgttctt caccaccagg gtggctgctg cctccgctgc tcagagcctc
     1261 cattgccaga tggtcgacaa gtggaccgtc accgaggctc ctgagcccag gcagctgctg
     1321 tggcaaaacc tgaacatcaa gctgttcagc aggatcatca ggcaatattt catttacttc
     1381 ttcgtggccg tgacaattct gttctacatg attcccatcg ccttcgtgtc cgctattacc
     1441 accctgaaaa acctccagag gatcattccc ttcatcaagc ctgtggtcga gattacagcc
     1501 atcaggacag tgctggagag cttcctgcct cagatcgccc tgatcgtgtt cctggccatg
     1561 ctgcccaagc tgctgctgtt cctgtccaag gccgagggca ttcctagcca atcccacgcc
     1621 atcagggctg ccagcggcaa gtatttctac ttcagcgtct tcaacgtgtt tatcggcgtg
     1681 accctggccg gaacactgtt caataccgtg aaggatattg ctaagaaccc caagctggac
     1741 atgatcatca atctgctcgc caccagcctg cctaaaagcg ccacattctt cctgacctat
     1801 gtcgccctca agttcttcat cggctacggc ctggagctga gcagaatcat ccccctgatc
     1861 attttccacc tgaaaaagaa gtatctgtgc aaaaccgagg ccgaagtgaa ggaggcttgg
     1921 taccctggcg acctgtccta cgccaccaga gtgcccggcg atatgctcat tctgaccatc
     1981 acattctgct acagcgtgat tgcccccctc atcctgatct tcggcatcac atacttcggc
     2041 ctgggatggc tggtgctgag gaatcaggcc ctgaaggtgt acgtccctag ctacgaatcc
     2101 tacggaagga tgtggcctca catccaccag agaatcctgg ccgccctgtt tctgtttcag
     2161 gtggtgatgt ttggctatct gggcgccaaa accttcttct acaccgccct cgtgatcccc
     2221 ctgatcatca ccagcctgat tttcggatac gtgtgcaggc agaagttcta cggcggcttc
     2281 gagcacaccg ctctggaggt ggcttgcaga gaactcaagc aatcccccga cctggaggag
     2341 atcttcagag cctacattcc tcacagcctg tccagccaca aacccgaaga gcacgagttc
     2401 aagggcgcca tgagcagata ccaggacttc aacgccatcg ccggcgtgtg accgcgggga
     2461 tccgcccctc tccctccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc
     2521 ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg
     2581 cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca
     2641 aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa
     2701 gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt
     2761 gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt
     2821 gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca
     2881 acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc
     2941 ggtacacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc
     3001 acggggacgt ggttttcctt tgaaaaacac gatgataata tggccacaac catggtgagc
     3061 aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg
     3121 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
     3181 gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg
     3241 gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac
     3301 atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
     3361 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc
     3421 atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag
     3481 aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag
     3541 ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag
     3601 accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag
     3661 ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag
     3721 ggccgccact ccaccggcgg catggacgag ctgtacaagt aagcggccgc gactctagat
     3781 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     3841 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     3901 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     3961 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa
     4021 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     4081 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     4141 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     4201 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     4261 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     4321 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     4381 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     4441 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     4501 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4561 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     4621 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     4681 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     4741 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     4801 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     4861 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     4921 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     4981 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     5041 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     5101 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     5161 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     5221 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     5281 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     5341 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     5401 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     5461 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     5521 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     5581 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     5641 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     5701 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     5761 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     5821 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5881 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5941 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     6001 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     6061 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     6121 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     6181 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     6241 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6301 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6361 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6421 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6481 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6541 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6601 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6661 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6721 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6781 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6841 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6901 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6961 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     7021 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     7081 tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     7141 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     7201 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     7261 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     7321 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     7381 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     7441 tagtcatcgc tattaccatg
//
