LOCUS       sequence-216385-        7121 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This construct was used to express mouse Otop2 in HEK-293 cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7121)
  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 7121)
  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: mOtop2_pcDNA3
FEATURES             Location/Qualifiers
     source          1..7121
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1785..1803)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1786..1803)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1823..1840)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1829..2053
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2099..2527
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2186..2205)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2396..2417
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2536..2556)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2541..2871
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2722..2857
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2784..2803
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2938..3732
                     /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(2992..3011)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3602..3621
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3906..4027
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3943..3962)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3997..4016
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4076..4092)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4076..4092)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4089..4111)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4100..4116
                     /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(4124..4130,4131..4148,4149..4154))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4169..4190
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4307..4324)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4478..5066)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4558..4577)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5237..6028,6029..6097))
                     /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     5860..5879
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6098..6202)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6277..6296)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6468..6847
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6848..7051
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7002..7022
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7096..7115
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7096..7114
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 aagcttggta ccgagctcgg atccactagt aacggccgcc agtgtgctgg aattcaccat
       61 gtcggaggaa ctggtcccgc atcccaatga gagcctgccg gggcctcggg ccagcccccg
      121 tgaggtgtgg aaaaagggcg gccgcctgct gtcggtgctg ttggctgtca acgtgctgct
      181 gctcgcctgc acgctcatca gcggcggtgc cttcaacaag gtggcggtgt atgacacgga
      241 cgtgttcgct ctgctcacca ccatgatgct gctggccgcg ctatggatag tcttctacct
      301 cctccgcacc gctcgctgtc cagacgcggt gccttaccgg gatgctcatg ctggtcccat
      361 ctggctccga ggtgggctgg tgctgtttgg aatctgcacc cttgtcatgg atgtcttcaa
      421 gactggctac tattccagtt tctttgagtg ccagtcggcc atcaagatcc tgcaccccat
      481 catccaagcc gtgtttgtca ttgtccagac ttactttctc tggatctccg ctaaagactg
      541 tatccatacc cacctggacc tgacccggtg tgggctcatg ttcaccctcg ccaccaacct
      601 cgccatctgg atggcagccg tagtggatga gtccgtgcac caggcccact cctacagtgg
      661 ttcccatggc aacaccagcc acacccgcct caaccctgac tcaaagcgtg caggaggtgc
      721 agctgaagaa gacccgtgcc tctgcagcac agccatctgt cagatcttcc agcaaggcta
      781 tttctacctg tatcccttca acatcgagta cagcctcttt gcttccacca tgctgtatgt
      841 catgtggaag aatgtgggaa ggctgctcgc ctccacccac ggccacggcc acacaccgtc
      901 cagggtcagc ctgttccggg agaccttctt tgccggccca gtcttgggcc tgctcctctt
      961 tgtggtgggg ttggctgtct tcatccttta tgaggtccag gtgagtggag agagaggcca
     1021 cacccgacag gccctggtca tctactacag ctttaacatt gtctgtctgg ggctcatgac
     1081 tctggtcagc ctgagtggtt cagtcatcta tcgttttgac cgtcgggcca tggaccatca
     1141 taagaacccg acacggaccc tggatgtggc cctgctaatg ggcgccgccc tgggccaata
     1201 cgctatctcc tactactcta tcgtggctgt ggtggtgggc tctcccagag acctgcaggg
     1261 ggcgctcaac ctgtcccacg ctctactcat gatcgctcag cacaccttcc agaatgtctt
     1321 catcattgag agcctccatc ggggacctcc tggggctgag cctcgtgaaa tgccccccaa
     1381 ggagccctgc caaggcatca cctttgctaa cctggatgct atccgcacct tgccttcctg
     1441 tccacccacc cccaggctgg tcattcccaa cctagaaagt cctcaggaag cagtggctat
     1501 catctcggcc cccaggtgcc actggcgacg tcggtgcctg aaagacatct ctctgtttct
     1561 cctgctctgt aacgtcatcc tgtggatcat gcctgccttt ggcgctcgcc cgcacttcag
     1621 caataccgtg gaggtggact tttacggtta ctccctctgg gcggccatcg tcaacatctg
     1681 cctacctttt ggcatcttct accgaatgca cgctgtctcc agtctactgg aggtctatgt
     1741 actgtcctga gcggccgctc gagcatgcat ctagagggcc ctattctata gtgtcaccta
     1801 aatgctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     1861 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     1921 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     1981 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     2041 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     2101 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2161 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2221 ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt
     2281 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2341 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2401 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2461 gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2521 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     2581 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     2641 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     2701 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     2761 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     2821 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     2881 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     2941 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3001 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     3061 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     3121 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     3181 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     3241 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     3301 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     3361 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     3421 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     3481 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     3541 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     3601 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     3661 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     3721 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     3781 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     3841 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     3901 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3961 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4021 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     4081 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4141 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4201 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4261 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4321 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4381 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4441 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4501 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4561 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4621 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac
     4681 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4741 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4801 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4861 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     4921 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4981 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5041 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5101 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5161 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5221 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5281 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5341 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5401 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5461 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5521 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5581 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5641 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5701 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5761 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5821 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5881 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5941 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6001 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6061 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6121 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6181 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     6241 cgggagatct cccgatcccc tatggtcgac tctcagtaca atctgctctg atgccgcata
     6301 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
     6361 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
     6421 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac attgattatt
     6481 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     6541 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     6601 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     6661 tcaatgggtg gactatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     6721 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     6781 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     6841 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
     6901 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
     6961 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
     7021 tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac ccactgctta
     7081 ctggcttatc gaaattaata cgactcacta tagggagacc c
//
