LOCUS       sequence-353942-        9084 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses full-length mouse TRPM5 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9084)
  AUTHORS   Hofmann T, Chubanov V, Gudermann T, Montell C
  TITLE     TRPM5 is a voltage-modulated and Ca(2+)-activated monovalent 
            selective cation channel.
  JOURNAL   Curr Biol. 2003 Jul 1;13(13):1153-8.
  PUBMED    12842017
REFERENCE   2  (bases 1 to 9084)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Curr 
            Biol."; date: "2003-07-1"; volume: "13(13)"; pages: "1153-8"
COMMENT     Sequence Label: pcDNA3.1-TRPM5
FEATURES             Location/Qualifiers
     source          1..9084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4348..4389
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4399..4416
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4439..4456)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4445..4669
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4715..5143
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4802..4821)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5012..5033
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5152..5172)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5157..5486
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5337..5472
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5399..5418
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5553..6347
                     /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(5607..5626)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6217..6236
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6523..6644
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6560..6579)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6614..6633
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6693..6709)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6693..6709)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6706..6728)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6717..6733
                     /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(6741..6747,6748..6765,6766..6771))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6786..6807
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6924..6941)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7095..7683)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7175..7194)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7854..8645,8646..8714))
                     /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     8477..8496
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8715..8819)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8894..8913)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagttaag cttggtaccg agctcggatc cactagtcca gtgtggtgga attgccctta
      721 ggcagtcctc atgcagtcca aggcactgac tgcctgactg gaaagccacc atgcaaacaa
      781 cccagagctc ctgccccggc agccccccag atactgagga tggctgggag cccatcctat
      841 gcaggggaga gatcaacttc ggagggtctg ggaagaagcg aggcaagttt gtgaaggtgc
      901 caagcagtgt ggccccctcc gtgctttttg aactcctgct caccgagtgg cacctgccag
      961 cccccaacct ggtggtgtcc ctggtgggtg aggaacgacc tttggctatg aagtcgtggc
     1021 ttcgggatgt cctgcgcaag gggctggtga aagcagctca gagcacaggt gcctggatcc
     1081 tgaccagtgc cctccacgtg ggcctggccc gccatgttgg acaagctgta cgtgatcact
     1141 ctctggctag cacatccacc aagatccgtg tagtggccat cggaatggcc tctctggatc
     1201 gaatccttca ccgtcaactt ctagatggtg tccaccaaaa ggaggatact cccatccact
     1261 acccagcaga tgagggcaac attcagggac ccctctgccc cctggacagc aatctctccc
     1321 acttcatcct tgtggagtca ggcgcccttg ggagtgggaa cgacgggctg acagagctgc
     1381 agctgagcct ggagaagcac atctctcagc agaggacagg ttatgggggc accagctgca
     1441 tccagatacc tgtcctttgc ctgttggtca atggtgaccc caacacccta gagaggattt
     1501 ccagggcagt ggagcaggct gccccatggc tgatcctggc aggttctggt ggcattgctg
     1561 atgtactcgc tgccctggtg agccagcctc atctcctggt gccccaggtg gctgagaagc
     1621 agttcagaga gaaattcccc agcgagtgtt tctcttggga agccattgta cactggacag
     1681 agctgttaca gaacattgct gcacaccccc acctgctcac agtatatgac ttcgagcagg
     1741 agggttcgga ggacctggac actgtcatcc tcaaggcact tgtgaaagcc tgcaagagcc
     1801 acagccaaga agcccaagac tacctagatg agctcaagtt agcagtggcc tgggatcgcg
     1861 tggacattgc caagagtgaa atcttcaatg gggacgtgga atggaagtcc tgtgacttgg
     1921 aagaggtgat gacagatgcc ctcgtgagca acaagcctga ctttgtccgc ctttttgtgg
     1981 acagcggtgc tgacatggcc gagttcttga cctatgggcg gctgcagcag ctttaccatt
     2041 ctgtgtcccc caagagcctc ctctttgaac tgctgcagcg taagcatgag gagggtaggc
     2101 tgacactggc cggcctgggt gcccagcagg ctcgggagct gcccattggt ctgcctgcct
     2161 tctcactcca cgaggtctcc cgcgtactca aagacttcct gcatgacgcc tgccgtggct
     2221 tctaccagga cgggcgcagg atggaggaga gagggccacc taagcggccc gcaggccaga
     2281 agtggctgcc agacctcagt aggaagagtg aagacccttg gagggacctg ttcctctggg
     2341 ctgtgctgca gaatcgttat gagatggcca catacttctg ggccatgggc cgggagggtg
     2401 tggctgctgc tctggctgcc tgcaagatca taaaggaaat gtcccacctg gagaaagagg
     2461 cagaggtggc ccgcaccatg cgtgaggcca agtatgagca gctggccctg gatcttttct
     2521 cagagtgcta cggcaacagt gaggaccgtg cctttgccct gctggtgcga aggaaccaca
     2581 gctggagcag gaccacgtgc ctgcacctgg ccactgaagc tgatgccaag gccttctttg
     2641 cccatgacgg tgtgcaagca ttcctgacca agatctggtg gggagacatg gccacaggca
     2701 cacccatcct acggcttctg ggtgccttca cctgcccagc cctcatctac acaaacctca
     2761 tctccttcag tgaggatgcc ccgcagagga tggacctaga agatctgcag gagccagaca
     2821 gcttggatat ggaaaagagc ttcctatgca gccggggtgg ccaattggag aagctaacag
     2881 aggcaccaag ggctccaggc gatctaggcc cacaagctgc cttcctgctc acacggtgga
     2941 ggaagttctg gggcgctcct gtgactgtgt tcctggggaa tgtggtcatg tacttcgcat
     3001 tcctcttcct gttcacctat gtcctgctgg tggacttcag gccaccaccc caggggccgt
     3061 ctggatccga ggttaccctc tatttctggg tgttcacact ggtgctggag gaaatccgac
     3121 agggcttctt cacagatgag gacacgcacc tggtgaagaa attcactctg tatgtggaag
     3181 acaactggaa caagtgtgac atggtggcca tcttcctgtt cattgtggga gtcacctgta
     3241 gaatggtgcc ctcggtgttt gaggctggca ggaccgttct ggccattgac ttcatggtgt
     3301 tcacacttcg gctcatccac atctttgcta ttcacaagca gttgggtcct aagatcatca
     3361 ttgtagagcg aatgatgaag gatgtcttct ttttcctctt cttcctgagc gtatggcttg
     3421 tggcctatgg tgtgaccact caggccctgc tgcatcccca tgatggccgt ttggagtgga
     3481 ttttccgccg tgtgctatac aggccttacc tgcagatctt tgggcaaatc cctctggatg
     3541 aaattgatga ggctcgtgtg aactgttctc ttcaccctct gctgctggaa agctcggctt
     3601 cctgccctaa tctctatgcc aactggctgg tcattctcct gctggttacc ttcctgcttg
     3661 tcactaatgt gctgctcatg aaccttctga tcgccatgtt cagctacaca ttccaggtgg
     3721 tgcaaggcaa tgcagacatg ttctggaagt ttcaacgcta ccacctcatc gttgaatacc
     3781 atggaagacc agctctggcc ccgcccttca tcctgctcag ccacctgagc ctggtgctca
     3841 agcaggtctt caggaaggaa gcccagcata agcgacaaca tctggagaga gacttgcctg
     3901 accccttgga ccagaagatc attacctggg aaacggttca aaaggagaac ttcctgagta
     3961 ccatggagaa acggaggagg gacagcgagg gggaggtgct gaggaaaacg gcacacagag
     4021 tggacttgat tgccaaatac atcggggggc tgagagagca agaaaagagg atcaagtgtc
     4081 tggaatcaca ggccaactac tgtatgctcc tcttgtcctc tatgacggat acactggctc
     4141 caggaggcac ctactcaagc tctcagaact gtggttgcag gagtcagcca gcctctgcta
     4201 gagacaggga gtacctagag tctggcttgc caccctctga cacctgaaat ggagaaacca
     4261 cttgctctag agccccagac caagggcaat tctgcagata tccagcacag tggcggccgc
     4321 tcgagtctag agggcccgcg gttcgaaggt aagcctatcc ctaaccctct cctcggtctc
     4381 gattctacgc gtaccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc
     4441 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     4501 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     4561 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     4621 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     4681 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     4741 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4801 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4861 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4921 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4981 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     5041 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     5101 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     5161 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5221 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     5281 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     5341 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5401 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     5461 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     5521 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     5581 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     5641 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     5701 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     5761 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     5821 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     5881 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5941 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     6001 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     6061 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     6121 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     6181 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     6241 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     6301 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     6361 gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     6421 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     6481 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     6541 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     6601 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     6661 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     6721 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     6781 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     6841 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     6901 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     6961 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     7021 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     7081 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     7141 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     7201 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     7261 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     7321 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     7381 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     7441 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     7501 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     7561 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7621 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7681 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7741 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7801 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7861 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7921 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7981 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     8041 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     8101 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     8161 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     8221 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     8281 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     8341 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     8401 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     8461 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     8521 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     8581 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     8641 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     8701 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8761 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8821 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
     8881 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
     8941 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
     9001 gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg
     9061 atgtacgggc cagatatacg cgtt
//
