LOCUS       sequence-221607-        7162 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 7162)
  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 7162)
  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..7162
                     /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..2912
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2763..2898
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2825..2844
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2979..3773
                     /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(3033..3052)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3643..3662
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3947..4068
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3984..4003)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4038..4057
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4117..4133)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4117..4133)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4130..4152)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4141..4157
                     /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(4165..4171,4172..4189,4190..4195))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4210..4231
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4348..4365)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4519..5107)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4599..4618)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5278..6069,6070..6138))
                     /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     5901..5920
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6139..6243)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6318..6337)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6509..6888
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6889..7092
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7043..7063
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7137..7156
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7137..7155
                     /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 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2341 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2401 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2461 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     2521 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     2581 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2641 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     2701 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     2761 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2821 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     2881 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     2941 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3001 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3061 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3121 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     3181 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     3241 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     3301 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     3361 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     3421 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     3481 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     3541 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     3601 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     3661 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     3721 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     3781 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     3841 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     3901 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     3961 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4021 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     4081 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4141 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4201 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4261 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4321 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4381 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4441 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4501 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4561 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4621 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4681 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4741 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4801 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4861 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4921 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4981 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5041 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5101 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5161 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5221 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5281 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5341 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5401 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5461 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5521 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5581 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5641 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5701 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5761 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5821 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5881 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5941 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6001 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6061 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6121 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6181 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6241 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     6301 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     6361 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     6421 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
     6481 cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa
     6541 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     6601 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     6661 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     6721 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     6781 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     6841 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     6901 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     6961 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     7021 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     7081 ataagcagag ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat
     7141 acgactcact atagggagac cc
//
