LOCUS       sequence-215298-        7960 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression plasmid for FLAG-tagged human T1R3 with signal 
            peptide of HLA class I histocompatibility antigen A-2 alpha chain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7960)
  AUTHORS   Park J, Selvam B, Sanematsu K, Shigemura N, Shukla D, Procko E
  TITLE     Structural architecture of a dimeric class C GPCR based on 
            co-trafficking of sweet taste receptor subunits.
  JOURNAL   J Biol Chem. 2019 Feb 5. pii: RA118.006173. doi: 
            10.1074/jbc.RA118.006173.
  PUBMED    30723160
REFERENCE   2  (bases 1 to 7960)
  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: "J Biol 
            Chem. 2019 Feb 5. pii: RA118.006173. doi: 10.1074/jbc.RA118.006173."
COMMENT     Sequence Label: pcDNA3.1(+)-HLA-Flag-natT1R3
FEATURES             Location/Qualifiers
     source          1..7960
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(38..55)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    44..268
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      314..742
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(401..420)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     611..632
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(751..771)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        756..1085
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      936..1071
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1152..1946
                     /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(1206..1225)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1816..1835
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2120..2241
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2157..2176)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2211..2230
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2290..2306)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2290..2306)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2303..2325)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2314..2330
                     /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(2338..2344,2345..2362,2363..2368))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2383..2404
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2521..2538)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2692..3277)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2772..2791)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3448..4239,4240..4308))
                     /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     4071..4090
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4309..4413)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4488..4507)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4679..5058
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5059..5262
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5213..5233
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5307..5326
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5307..5325
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      5345..5354
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5429..5452
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
       61 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
      121 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
      181 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
      241 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
      301 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
      361 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
      421 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
      481 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
      541 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
      601 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
      661 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
      721 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
      781 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
      841 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
      901 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
      961 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     1021 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     1081 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     1141 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     1201 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     1261 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     1321 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     1381 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     1441 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     1501 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     1561 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     1621 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     1681 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     1741 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     1801 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     1861 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     1921 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     1981 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     2041 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     2101 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     2161 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     2221 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     2281 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     2341 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     2401 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     2461 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     2521 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     2581 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2641 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2701 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2761 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2821 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2881 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2941 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3001 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3061 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3121 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3181 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg
     3241 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     3301 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     3361 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3421 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3481 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3541 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     3601 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     3661 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     3721 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     3781 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     3841 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     3901 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     3961 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4021 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4081 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4141 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     4201 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4261 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4321 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4381 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     4441 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
     4501 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     4561 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
     4621 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
     4681 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     4741 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     4801 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     4861 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     4921 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     4981 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     5041 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     5101 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     5161 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     5221 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa
     5281 cccactgctt actggcttat cgaaattaat acgactcact atagggagac ccaagctggc
     5341 tagcgccacc atggccgtca tggcgccccg aaccctcgtc ctgctactct cgggggccct
     5401 ggccctgacc cagacctggg cgggctccga ctacaaggac gatgatgaca agggaggatc
     5461 cgccccattg tgcctgtcac agcaacttag gatgaagggg gactacgtgc tgggggggct
     5521 gttccccctg ggcgaggccg aggaggctgg cctccgcagc cggacacggc ccagcagccc
     5581 tgtgtgcacc aggttctcct caaacggcct gctctgggca ctggccatga aaatggccgt
     5641 ggaggagatc aacaacaagt cggatctgct gcccgggctg cgcctgggct acgacctctt
     5701 tgatacgtgc tcggagcctg tggtggccat gaagcccagc ctcatgttcc tggccaaggc
     5761 aggcagccgc gacatcgccg cctactgcaa ctacacgcag taccagcccc gtgtgctggc
     5821 tgtcatcggg ccccactcgt cagagctcgc catggtcacc ggcaagttct tcagcttctt
     5881 cctcatgccc caggtcagct acggtgctag catggagctg ctgagcgccc gggagacctt
     5941 cccctccttc ttccgcaccg tgcccagcga ccgtgtgcag ctgacggccg ccgcggagct
     6001 gctgcaggag ttcggctgga actgggtggc cgccctgggc agcgacgacg agtacggccg
     6061 gcagggcctg agcatcttct cggccctggc cgcggcacgc ggcatctgca tcgcgcacga
     6121 gggcctggtg ccgctgcccc gtgccgatga ctcgcggctg gggaaggtgc aggacgtcct
     6181 gcaccaggtg aaccagagca gcgtgcaggt ggtgctgctg ttcgcctccg tgcacgccgc
     6241 ccacgccctc ttcaactaca gcatcagcag caggctctcg cccaaggtgt gggtggccag
     6301 cgaggcctgg ctgacctctg acctggtcat ggggctgccc ggcatggccc agatgggcac
     6361 ggtgcttggc ttcctccaga ggggtgccca gctgcacgag ttcccccagt acgtgaagac
     6421 gcacctggcc ctggccaccg acccggcctt ctgctctgcc ctgggcgaga gggagcaggg
     6481 tctggaggag gacgtggtgg gccagcgctg cccgcagtgt gactgcatca cgctgcagaa
     6541 cgtgagcgca gggctaaatc accaccagac gttctctgtc tacgcagctg tgtatagcgt
     6601 ggcccaggcc ctgcacaaca ctcttcagtg caacgcctca ggctgccccg cgcaggaccc
     6661 cgtgaagccc tggcagctcc tggagaacat gtacaacctg accttccacg tgggcgggct
     6721 gccgctgcgg ttcgacagca gcggaaacgt ggacatggag tacgacctga agctgtgggt
     6781 gtggcagggc tcagtgccca ggctccacga cgtgggcagg ttcaacggca gcctcaggac
     6841 agagcgcctg aagatccgct ggcacacgtc tgacaaccag aagcccgtgt cccggtgctc
     6901 gcggcagtgc caggagggcc aggtgcgccg ggtcaagggg ttccactcct gctgctacga
     6961 ctgtgtggac tgcgaggcgg gcagctaccg gcaaaaccca gacgacatcg cctgcacctt
     7021 ttgtggccag gatgagtggt ccccggagcg aagcacacgc tgcttccgcc gcaggtctcg
     7081 gttcctggca tggggcgagc cggctgtgct gctgctgctc ctgctgctga gcctggcgct
     7141 gggccttgtg ctggctgctt tggggctgtt cgttcaccat cgggacagcc cactggttca
     7201 ggcctcgggg gggcccctgg cctgctttgg cctggtgtgc ctgggcctgg tctgcctcag
     7261 cgtcctcctg ttccctggcc agcccagccc tgcccgatgc ctggcccagc agcccttgtc
     7321 ccacctcccg ctcacgggct gcctgagcac actcttcctg caggcggccg agatcttcgt
     7381 ggagtcagaa ctgcctctga gctgggcaga ccggctgagt ggctgcctgc gggggccctg
     7441 ggcctggctg gtggtgctgc tggccatgct ggtggaggtc gcactgtgca cctggtacct
     7501 ggtggccttc ccgccggagg tggtgacgga ctggcacatg ctgcccacgg aggcgctggt
     7561 gcactgccgc acacgctcct gggtcagctt cggcctagcg cacgccacca atgccacgct
     7621 ggcctttctc tgcttcctgg gcactttcct ggtgcggagc cagccgggcc gctacaaccg
     7681 tgcccgtggc ctcacctttg ccatgctggc ctacttcatc acctgggtct cctttgtgcc
     7741 cctcctggcc aatgtgcagg tggtcctcag gcccgccgtg cagatgggcg ccctcctgct
     7801 ctgtgtcctg ggcatcctgg ctgccttcca cctgcccagg tgttacctgc tcatgcggca
     7861 gccagggctc aacacccccg agttcttcct gggagggggc cctggggatg cccaaggcca
     7921 gaatgacggg aacacaggaa atcaggggaa acatgagtga
//
