LOCUS       sequence-292301-        8023 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for c-myc-tagged mGluR3 C127A and I578A
            mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8023)
  TITLE     Mammalian expression plasmids for production of metabotropic 
            glutamate receptor type 3
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8023)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pcDNA3_HLA-cmyc-GRM3 C127A TMD I578A
FEATURES             Location/Qualifiers
     source          1..8023
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      901..910
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             985..1014
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3614..3631)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3620..3844
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3890..4318
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3977..3996)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4187..4208
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4327..4347)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4332..4661
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4512..4647
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4574..4593
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4728..5522
                     /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(4782..4801)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5392..5411
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5696..5817
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5733..5752)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5787..5806
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5866..5882)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5866..5882)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5879..5901)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5890..5906
                     /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(5914..5920,5921..5938,5939..5944))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5959..5980
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6097..6114)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6268..6856)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6348..6367)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7027..7818,7819..7887))
                     /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     7650..7669
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7888..7992)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gccaccatgg ccgtcatggc gccccgaacc ctcgtcctgc tactctcggg ggccctggcc
      961 ctgacccaga cctgggcggg ctccgaacaa aagctcattt ctgaagagga cttgggagga
     1021 tccaggagag agattaaaat agaaggtgac cttgttttag ggggcctgtt tcctattaac
     1081 gaaaaaggca ctggaactga agaatgtggg cgaatcaatg aagaccgagg gattcaacgc
     1141 ctggaagcca tgttgtttgc tattgatgaa atcaacaaag atgattactt gctaccagga
     1201 gtgaagttgg gtgttcacat tttggataca tgttcaaggg atacctatgc attggagcaa
     1261 tcactggagt ttgtcagggc atctttgaca aaagtggatg aagctgagta tatggctcct
     1321 gatggatcct atgccattca agaaaacatc ccacttctca ttgcaggggt cattggtggc
     1381 tcttatagca gtgtttccat acaggtggca aacctgctgc ggctcttcca gatccctcag
     1441 atcagctacg catccaccag cgccaaactc agtgataagt cgcgctatga ttactttgcc
     1501 aggaccgtgc cccccgactt ctaccaggcc aaagccatgg ctgagatctt gcgcttcttc
     1561 aactggacct acgtgtccac agtagcctcc gagggtgatt acggggagac agggatcgag
     1621 gccttcgagc aggaagcccg cctgcgcaac atctgcatcg ctacggcgga gaaggtgggc
     1681 cgctccaaca tccgcaagtc ctacgacagc gtgatccgag aactgttgca gaagcccaac
     1741 gcgcgcgtcg tggtcctctt catgcgcagc gacgactcgc gggagctcat tgcggccgcc
     1801 agccgcgcca atgcctcctt cacctgggtg gccagcgacg gctggggcgc gcaggagagc
     1861 atcatcaagg gcagcgagca tgtggcctac ggcgccatca ccctggagct ggcctcccag
     1921 cctgtccgcc agttcgaccg ctacttccag agcctcaacc cctacaacaa ccaccgcaac
     1981 ccctggttcc gggacttctg ggagcaaaag tttcagtgca gcctccagaa caaacgcaac
     2041 cacaggcgcg tctgcgacaa gcacctggcc atcgacagca gcaactacga gcaagagtcc
     2101 aagatcatgt ttgtggtgaa cgcggtgtat gccatggccc acgctttgca caaaatgcag
     2161 cgcaccctct gtcccaacac taccaagctt tgtgatgcta tgaagatcct ggatgggaag
     2221 aagttgtaca aggattactt gctgaaaatc aacttcacgg ctccattcaa cccaaataaa
     2281 gatgcagata gcatagtcaa gtttgacact tttggagatg gaatggggcg atacaacgtg
     2341 ttcaatttcc aaaatgtagg tggaaagtat tcctacttga aagttggtca ctgggcagaa
     2401 accttatcgc tagatgtcaa ctctatccac tggtcccgga actcagtccc cacttcccag
     2461 tgcagcgacc cctgtgcccc caatgaaatg aagaatatgc aaccagggga tgtctgctgc
     2521 tggatttgca tcccctgtga accgtacgaa tacctggctg atgagtttac ctgtatggat
     2581 tgtgggtctg gacagtggcc cactgcagac ctaactggat gctatgacct tcctgaggac
     2641 tacatcaggt gggaagacgc ctgggccgct ggcccagtca ccattgcctg tctgggtttt
     2701 atgtgtacat gcatggttgt aactgttttt atcaagcaca acaacacacc cttggtcaaa
     2761 gcatcgggcc gagaactctg ctacatctta ttgtttgggg ttggcctgtc atactgcatg
     2821 acattcttct tcattgccaa gccatcacca gtcatctgtg cattgcgccg actcgggctg
     2881 gggagttcct tcgctatctg ttactcagcc ctgctgacca agacaaactg cattgcccgc
     2941 atcttcgatg gggtcaagaa tggcgctcag aggccaaaat tcatcagccc cagttctcag
     3001 gttttcatct gcctgggtct gatcctggtg caaattgtga tggtgtctgt gtggctcatc
     3061 ctggaggccc caggcaccag gaggtatacc cttgcagaga agcgggaaac agtcatccta
     3121 aaatgcaatg tcaaagattc cagcatgttg atctctctta cctacgatgt gatcctggtg
     3181 atcttatgca ctgtgtacgc cttcaaaacg cggaagtgcc cagaaaattt caacgaagct
     3241 aagttcatag gttttaccat gtacaccacg tgcatcatct ggttggcctt cctccctata
     3301 ttttatgtga catcaagtga ctacagagtg cagacgacaa ccatgtgcat ctctgtcagc
     3361 ctgagtggct ttgtggtctt gggctgtttg tttgcaccca aggttcacat catcctgttt
     3421 caaccccaga agaatgttgt cacacacaga ctgcacctca acaggttcag tgtcagtgga
     3481 actgggacca catactctca gtcctctgca agcacgtatg tgccaacggt gtgcaatggg
     3541 cgggaagtcc tcgactccac cacctcatct ctgtgactcg agtctagagg gcccgtttaa
     3601 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     3661 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     3721 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     3781 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     3841 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     3901 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3961 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4021 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4081 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4141 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4201 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     4261 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     4321 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4381 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     4441 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     4501 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     4561 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     4621 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     4681 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     4741 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     4801 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4861 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     4921 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4981 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     5041 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     5101 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     5161 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     5221 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     5281 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     5341 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     5401 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     5461 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     5521 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5581 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5641 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     5701 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5761 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5821 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     5881 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5941 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     6001 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     6061 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6121 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6181 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6241 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6301 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6361 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6421 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6481 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6541 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6601 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6661 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     6721 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6781 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6841 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6901 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6961 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7021 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7081 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7141 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7201 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     7261 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7321 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7381 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7441 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7501 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7561 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7621 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7681 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7741 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7801 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7861 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7921 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7981 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
