LOCUS       sequence-221605-        7231 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This construct was used to express mouse Otop1 in HEK-293 cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7231)
  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 7231)
  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: mOtop1_pcDNA3
FEATURES             Location/Qualifiers
     source          1..7231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1896..1914)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1897..1914)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1934..1951)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1940..2164
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2210..2638
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2297..2316)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2507..2528
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2647..2667)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2652..2981
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2832..2967
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2894..2913
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3048..3842
                     /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(3102..3121)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3712..3731
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4016..4137
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4053..4072)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4107..4126
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4186..4202)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4186..4202)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4199..4221)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4210..4226
                     /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(4234..4240,4241..4258,4259..4264))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4279..4300
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4417..4434)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4588..5176)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4668..4687)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5347..6138,6139..6207))
                     /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     5970..5989
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6208..6312)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6387..6406)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6578..6957
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6958..7161
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7112..7132
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7206..7225
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7206..7224
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 aagcttggta ccgagctcgg atccactagt aacggccgcc agtgtgctgg aattcaccat
       61 gcctgggggc ccgggcgcgc cctcttctcc cgcagcatcc tcaggctcct cgcgtgccgc
      121 gccgtcgggg atcgcggcat gccctctgtc cccacctccc ctagcccgag ggtctccaca
      181 agcttccggt ccccggcgcg gcgccagcgt cccgcagaag ctggcggaga ctctgagcag
      241 ccagtatgga ctgaacgtgt tcgtggcggg gctgctgttc ctgctggcct gggcggtgca
      301 cgcgacgggt gtgggcaaga gcgacctgct gtgcgttcta accgcgctta tgctgctgca
      361 gctgctctgg atgctgtggt acgtgggtcg cagctacatg cagcgccgcc tcatccgccc
      421 caaggacacg cacgcgggtg cgcgctggct tcgcggaagc atcacgttgt tcgcgtttat
      481 cactgtcgtc ctgggatgct tgaaagtcgc atacttcatt ggattctcgg agtgcttgtc
      541 agccaccgag ggagttttcc cagtcaccca tgcagtgcat accctattgc aggtgtattt
      601 cctctggggc catgctaagg atatcatcat gtctttcaaa acactggaaa ggtttggggt
      661 gatccattca gtgttcacga acctcctact gtgggccaac agcgtcctga atgaatcaaa
      721 gcaccagctg aatgagcaca aggaacggct gatcactctg ggctttggca acatcaccat
      781 cgttttggat gaccacacac cacagtgtaa ctgcacacca cccgccctct gctctgccct
      841 ctcccatggg atttactatc tgtacccctt caacattgag taccagatcc tggcctcgac
      901 catgctctac gtgctgtgga agaacatcgg gcgcagagtg gacagctccc agcaccagaa
      961 gatgcagtgc agattcgacg gggtcctagt gggctccgtg ctgggcttga cagtgctggc
     1021 tgccaccatc gccgtggttg tggtgtacat gatccacatc gggcgctcca aatccaagag
     1081 cgagtcggct ctcatcatgt tctatttgta cgctatcacg gtgctgctgc ttatgggggc
     1141 cgcagggcta gtcggaagct ggatttacag ggtggatgag aagtctctgg atgagtcaaa
     1201 gaacccagcg cgcaagctgg atgttgacct gttggtggcc accgcctccg gctcctggct
     1261 cctctcctgg ggctccattc tggccatcgc ctgtgctgag actcgcccac cgtacacctg
     1321 gtacaacctg ccctactcgg tcctggtgat cgtggagaag tatgtccaaa acattttcat
     1381 catcgagtcc gtgcacctcg agcctgaggg ggtcccggag gatgtgcgca ctctgcgtgt
     1441 ggtcaccgtc tgcagcagcg aggctgccgc actggctgca tccactctcg ggagccaggg
     1501 gatggcccag gatgggtcac ctgctgtcaa tggaaatctg tgtctgcagc agaggtgtgg
     1561 gaaagaggac caggagtctg gctgggaagg agctacgggg acaacccgat gtctggactt
     1621 ccttcagggc ggcatgaaga ggaggcttct cagaaacatc acggcctttc tgtttctttg
     1681 caacatctcg ctttggattc cccctgcctt tggctgccgt cccgagtatg acaacggatt
     1741 ggaggaaatt gtctttggct ttgaaccctg gataattgtg gtcaacctgg ccatgccctt
     1801 ttccattttc taccggatgc acgcagctgc tgccctcttt gaggtctatt gtaagatcta
     1861 ggcggccgct cgagcatgca tctagagggc cctattctat agtgtcacct aaatgctaga
     1921 gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     1981 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     2041 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     2101 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     2161 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     2221 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     2281 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     2341 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     2401 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     2461 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     2521 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     2581 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     2641 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2701 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     2761 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     2821 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     2881 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     2941 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     3001 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     3061 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     3121 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     3181 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     3241 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     3301 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     3361 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     3421 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     3481 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     3541 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     3601 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     3661 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     3721 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     3781 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     3841 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     3901 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     3961 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     4021 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     4081 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     4141 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     4201 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4261 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4321 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4381 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4441 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4501 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4561 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4621 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4681 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4741 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4801 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4861 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4921 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4981 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     5041 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5101 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5161 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5221 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5281 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5341 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5401 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5461 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5521 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5581 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5641 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5701 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5761 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5821 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5881 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5941 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6001 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6061 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6121 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6181 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6241 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6301 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     6361 cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     6421 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     6481 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt
     6541 gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat
     6601 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     6661 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     6721 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     6781 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     6841 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     6901 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
     6961 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
     7021 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
     7081 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
     7141 gaggtctata taagcagagc tctctggcta actagagaac ccactgctta ctggcttatc
     7201 gaaattaata cgactcacta tagggagacc c
//
