LOCUS       sequence-216384-        7232 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 7232)
  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 7232)
  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..7232
                     /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..2982
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2833..2968
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2895..2914
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3049..3843
                     /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(3103..3122)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3713..3732
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4017..4138
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4054..4073)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4108..4127
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4187..4203)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4187..4203)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4200..4222)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4211..4227
                     /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(4235..4241,4242..4259,4260..4265))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4280..4301
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4418..4435)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4589..5177)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4669..4688)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5348..6139,6140..6208))
                     /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     5971..5990
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6209..6313)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6388..6407)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6579..6958
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6959..7162
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7113..7133
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7207..7226
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7207..7225
                     /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 accggctccg 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 tcggggcatc 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 gggatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     2641 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca ggcaggcaga
     2701 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     2761 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     2821 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     2881 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     2941 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     3001 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     3061 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3121 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3181 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     3241 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3301 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3361 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     3421 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     3481 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     3541 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     3601 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     3661 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     3721 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     3781 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     3841 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     3901 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     3961 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     4021 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4081 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     4141 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     4201 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4261 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4321 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4381 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4441 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4501 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4561 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4621 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4681 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4741 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt
     4801 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4861 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4921 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4981 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     5041 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5101 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5161 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5221 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5281 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5341 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5401 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5461 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5521 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5581 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5641 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5701 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5761 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5821 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5881 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5941 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6001 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6061 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6121 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6181 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6241 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6301 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     6361 tcccgatccc ctatggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca
     6421 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     6481 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
     6541 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta
     6601 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     6661 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     6721 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     6781 ggactattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     6841 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     6901 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     6961 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     7021 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     7081 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     7141 ggaggtctat ataagcagag ctctctggct aactagagaa cccactgctt actggcttat
     7201 cgaaattaat acgactcact atagggagac cc
//
