LOCUS       sequence-221622-        7120 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 7120)
  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 7120)
  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..7120
                     /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..2870
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2721..2856
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2783..2802
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2937..3731
                     /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(2991..3010)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3601..3620
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3905..4026
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3942..3961)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3996..4015
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4075..4091)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4075..4091)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4088..4110)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4099..4115
                     /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(4123..4129,4130..4147,4148..4153))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4168..4189
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4306..4323)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4477..5065)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4557..4576)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5236..6027,6028..6096))
                     /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     5859..5878
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6097..6201)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6276..6295)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6467..6846
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6847..7050
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7001..7021
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7095..7114
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7095..7113
                     /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 cgggggctcc ctttagggtt ccgatttagt
     2281 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2341 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2401 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2461 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2521 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     2581 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     2641 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     2701 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     2761 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     2821 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     2881 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     2941 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3001 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3061 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     3121 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3181 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3241 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3301 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3361 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3421 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     3481 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3541 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     3601 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     3661 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     3721 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     3781 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     3841 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     3901 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3961 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4021 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     4081 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4141 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4201 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4261 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4321 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4381 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4441 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4501 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4561 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4621 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4681 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4741 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4801 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4861 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4921 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4981 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5041 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5101 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5161 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5221 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5281 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5341 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5401 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5461 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5521 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5581 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5641 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5701 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5761 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5821 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5881 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5941 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6001 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6061 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6121 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6181 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     6241 gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag
     6301 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     6361 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
     6421 aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg
     6481 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     6541 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     6601 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     6661 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     6721 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     6781 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     6841 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
     6901 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     6961 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
     7021 gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc cactgcttac
     7081 tggcttatcg aaattaatac gactcactat agggagaccc
//
