LOCUS       sequence-216386-        7163 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This construct was used to express mouse Otop3 in HEK-293 cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7163)
  AUTHORS   Tu YH, Cooper AJ, Teng B, Chang RB, Artiga DJ, Turner HN, Mulhall 
            EM, Ye W, Smith AD, Liman ER
  TITLE     An evolutionarily conserved gene family encodes proton-selective ion
            channels.
  JOURNAL   Science. 2018 Mar 2;359(6379):1047-1050. doi: 
            10.1126/science.aao3264. Epub 2018 Jan 25.
  PUBMED    29371428
REFERENCE   2  (bases 1 to 7163)
  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"; doi: 
            "10.1126/science.aao3264"; journalName: "Science"; date: "2018-03-2-
            2"; volume: "359"; issue: "6379"; pages: "1047-1050"
COMMENT     Sequence Label: mOtop3_pcDNA3
FEATURES             Location/Qualifiers
     source          1..7163
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1827..1845)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1828..1845)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1865..1882)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1871..2095
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2141..2569
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2228..2247)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2438..2459
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2578..2598)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2583..2913
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2764..2899
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2826..2845
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2980..3774
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3034..3053)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3644..3663
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3948..4069
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3985..4004)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4039..4058
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4118..4134)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4118..4134)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4131..4153)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4142..4158
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4166..4172,4173..4190,4191..4196))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4211..4232
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4349..4366)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4520..5108)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4600..4619)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5279..6070,6071..6139))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5902..5921
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6140..6244)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6319..6338)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6510..6889
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6890..7093
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7044..7064
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7138..7157
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7138..7156
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 aagcttggta ccgagctcgg atccactagt aacggccgcc agtgtgctgg aattcaccat
       61 ggcttcgcag acttcagccc ctgctgagcc tgcacccatg ccttctccag aagcaaagac
      121 cactgaggga gcatcctcct atgaccaagc agacatggag actaagcatg cggggtcccc
      181 atgccccccg aagcagaagt cctggctggc tcggcatttc tccctgctgt tgcggagaga
      241 ccgacaggcc cagaaggccg ggcagctctt ctccgggctc ctggctctca atgtggtatt
      301 cctgggtggg gcttttatct gtagcatgat attcaacaag gtgtctgtca ctctggggga
      361 cgtgtggatc ctgctggctg cactgaaggt cctctccctg ttgtggctcc tctactacac
      421 agtgggtacc acccggaagc cccacgcagt cctgtaccgt gatccccacg cgggacccat
      481 ctgggtgagg ggctccctgg tgctcttcgg cagctgtacc gtctgcctga acatatttcg
      541 aatgggttat gacgtgagcc acattcactg caagtcagaa gtggaactca tctttccggc
      601 cattgagatt gtctttatga ttattcagac atgggtgctc tggagacact gtaaggactg
      661 tgttcaggtg cagaccaact tcaccaggtg tggactcatg ctgactctgg ccacaaactt
      721 gctgatgtgg gttttggctg tgaccaatga ctccatgcac cgtgagattg aggctgagct
      781 cgacgccctc atggagaagt tctcaggtaa cgggaccaac acttgcatgt gcctcaacac
      841 cacagtgtgc gaggtcttcc ggaagggcta tctgatgctc taccccttca gcactgagta
      901 ctgcctcatc tgctgtgctg tgctcttcgt catgtggaag aacgtgagcc gcagcctggc
      961 agcccacaca ggtgcccatc ccaacagatc acccttccgc ctgcacggga ccatctttgg
     1021 gccactgctg ggccttttgg cactggtggc tggcgtatgt gtctttgtgc tcttccaaat
     1081 cgaagcgagt ggccctgaca tagcccgcca gtacttcacc ctctactatg cgttctatgt
     1141 ggctgtgctg cccaccatga gcctggcgtg cctggctggt acagccatcc atgggctgga
     1201 ggagcgcgaa ttggacacgc tcaaaaaccc cacacgcagc ctggatgtgg tgctgttgat
     1261 gggcgctgca ctgggccaga tgggcatcgc ctacttctcc atcgtggcta tcgtggccac
     1321 ccagccgcac gagctactca accagcttat cctggcctac tcactgctgc taatcctgca
     1381 gcacatcact cagaatctct tcatcatcga gggcctgcac cgccgccccc tctgggagcc
     1441 tgcggtctca ggtgtgatgg agaagcagga tgttgagcta ccccgcagag gctccctgcg
     1501 ggagctgggt caggacctac ggagagcctc aagggcctac atccactcct tcagccacct
     1561 caactggaaa cgcaggatgc tcaaggagat ctctctcttc ctcatcctct gcaacatcac
     1621 actgtggatg atgcccgcct tcggcataca cccagagttt gagaacgggc tggaaaagga
     1681 tttttacggc tacaggacct ggttcaccat tgttaacttc ggcctgcccc tgggtgtgtt
     1741 ctaccgcatg cactctgtcg gggggctggt ggaggtctac ctgggggcct gagcggccgc
     1801 tcgagcatgc atctagaggg ccctattcta tagtgtcacc taaatgctag agctcgctga
     1861 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     1921 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     1981 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     2041 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     2101 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     2161 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2221 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2281 caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2341 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2401 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2461 acaacactca accctatctc ggtctattct tttgatttat aagggatttt ggggatttcg
     2521 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     2581 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc aggcaggcag aagtatgcaa
     2641 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     2701 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     2761 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     2821 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     2881 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     2941 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     3001 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     3061 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     3121 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     3181 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     3241 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     3301 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     3361 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     3421 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     3481 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     3541 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     3601 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     3661 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     3721 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     3781 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     3841 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     3901 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     3961 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     4021 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     4081 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     4141 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     4201 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     4261 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     4321 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     4381 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     4441 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4501 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4561 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4621 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4681 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt
     4741 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4801 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4861 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4921 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     4981 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5041 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5101 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     5161 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     5221 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     5281 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     5341 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     5401 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5461 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5521 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5581 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5641 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5701 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5761 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5821 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5881 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5941 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6001 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6061 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6121 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     6181 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     6241 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
     6301 cctatggtcg actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
     6361 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
     6421 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct
     6481 tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta
     6541 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     6601 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     6661 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggactattt
     6721 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     6781 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     6841 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     6901 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     6961 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     7021 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     7081 tataagcaga gctctctggc taactagaga acccactgct tactggctta tcgaaattaa
     7141 tacgactcac tatagggaga ccc
//
