LOCUS       sequence-224107-        7924 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression plasmid for FLAG-tagged human T1R2 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 7924)
  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 7924)
  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-natT1R2
FEATURES             Location/Qualifiers
     source          1..7924
                     /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"
     primer_bind     998..1017
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     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..3280)
                     /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(3451..4242,4243..4311))
                     /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     4074..4093
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4312..4416)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4491..4510)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4682..5061
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5062..5265
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5216..5236
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5310..5329
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5310..5328
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      5348..5357
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5432..5455
                     /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 ggtagcggtg gtttttttgt
     3241 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3301 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3361 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3421 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3481 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3541 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3601 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3661 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3721 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3781 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3841 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     3901 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3961 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4021 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4081 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4141 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4201 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4261 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4321 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4381 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4441 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
     4501 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     4561 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     4621 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt
     4681 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
     4741 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     4801 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     4861 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     4921 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     4981 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     5041 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
     5101 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     5161 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
     5221 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga
     5281 gaacccactg cttactggct tatcgaaatt aatacgactc actataggga gacccaagct
     5341 ggctagcgcc accatggccg tcatggcgcc ccgaaccctc gtcctgctac tctcgggggc
     5401 cctggccctg acccagacct gggcgggctc cgactacaag gacgatgatg acaagggagg
     5461 atccgctgag aactcggact tctacctgcc tggggattac ctcctgggtg gcctcttctc
     5521 cctccatgcc aacatgaagg gcattgttca ccttaacttc ctgcaggtgc ccatgtgcaa
     5581 ggagtatgaa gtgaaggtga taggctacaa cctcatgcag gccatgcgct ttgcggtgga
     5641 ggagatcaac aatgacagca gcctgctgcc tggtgtgctg ctgggctatg agatcgtgga
     5701 tgtgtgctac atctccaaca atgtccagcc ggtgctctac ttcctggcac acgaggacaa
     5761 cctccttccc atccaagagg actacagtaa ctacatttcc cgtgtggtgg ctgtcattgg
     5821 ccctgacaac tccgagtctg tcatgactgt ggccaacttc ctctccctat ttctccttcc
     5881 acagatcacc tacagcgcca tcagcgatga gctgcgagac aaggtgcgct tcccggcttt
     5941 gctgcgtacc acacccagcg ccgaccacca catcgaggcc atggtgcagc ttatgctgca
     6001 cttccgctgg aactggatca ttgtgctggt gagcagcgac acctatggcc gcgacaatgg
     6061 ccagctgctt ggcgagcgcg tggcccggcg cgacatctgc atcgccttcc aggagacgct
     6121 gcccacactg cagcccaacc agaacatgac gtcagaggag cgccagcgcc tggtgaccat
     6181 tgtggacaag ctgcagcaga gcacagcgcg cgtcgtggtc gtgttctcgc ccgacctgac
     6241 cctgtaccac ttcttcaatg aggtgctgcg ccagaacttc actggcgccg tgtggatcgc
     6301 ctccgagtcc tgggccatcg acccggtcct gcacaacctc acggagctgc gccacttggg
     6361 caccttcctg ggcatcacca tccagagcgt gcccatcccg ggcttcagtg agttccgcga
     6421 gtggggccca caggctgggc cgccacccct cagcaggacc agccagagct atacctgcaa
     6481 ccaggagtgc gacaactgcc tgaacgccac cttgtccttc aacaccattc tcaggctctc
     6541 tggggagcgt gtcgtctaca gcgtgtactc tgcggtctat gctgtggccc atgccctgca
     6601 cagcctcctc ggctgtgaca aaagcacctg caccaagagg gtggtctacc cctggcagct
     6661 gcttgaggag atctggaagg tcaacttcac tctcctggac caccaaatct tcttcgaccc
     6721 gcaaggggac gtggctctgc acttggagat tgtccagtgg caatgggacc ggagccagaa
     6781 tcccttccag agcgtcgcct cctactaccc cctgcagcga cagctgaaga acatccaaga
     6841 catctcctgg cacaccatca acaacacgat ccctatgtcc atgtgttcca agaggtgcca
     6901 gtcagggcaa aagaagaagc ctgtgggcat ccacgtctgc tgcttcgagt gcatcgactg
     6961 ccttcccggc accttcctca accacactga agatgaatat gaatgccagg cctgcccgaa
     7021 taacgagtgg tcctaccaga gtgagacctc ctgcttcaag cggcagctgg tcttcctgga
     7081 atggcatgag gcacccacca tcgctgtggc cctgctggcc gccctgggct tcctcagcac
     7141 cctggccatc ctggtgatat tctggaggca cttccagaca cccatagttc gctcggctgg
     7201 gggccccatg tgcttcctga tgctgacact gctgctggtg gcatacatgg tggtcccggt
     7261 gtacgtgggg ccgcccaagg tctccacctg cctctgccgc caggccctct ttcccctctg
     7321 cttcacaatc tgcatctcct gtatcgccgt gcgttctttc cagatcgtct gcgccttcaa
     7381 gatggccagc cgcttcccac gcgcctacag ctactgggtc cgctaccagg ggccctacgt
     7441 ctctatggca tttatcacgg tactcaaaat ggtcattgtg gtaattggca tgctggccac
     7501 gggcctcagt cccaccaccc gtactgaccc cgatgacccc aagatcacaa ttgtctcctg
     7561 taaccccaac taccgcaaca gcctgctgtt caacaccagc ctggacctgc tgctctcagt
     7621 ggtgggtttc agcttcgcct acatgggcaa agagctgccc accaactaca acgaggccaa
     7681 gttcatcacc ctcagcatga ccttctattt cacctcatcc gtctccctct gcaccttcat
     7741 gtctgcctac agcggggtgc tggtcaccat cgtggacctc ttggtcactg tgctcaacct
     7801 cctggccatc agcctgggct acttcggccc caagtgctac atgatcctct tctacccgga
     7861 gcgcaacacg cccgcctact tcaacagcat gatccagggc tacaccatga ggagggacta
     7921 gtga
//
