LOCUS       sequence-224121-        5654 bp    DNA     linear   UNA 14-APR-2025
DEFINITION  mammalian expression plasmid for FLAG-tagged human T1R2 with signal 
            peptide of influenza hemagglutinin; expression-enhanced variant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5654)
  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 5654)
  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: "J Biol 
            Chem. 2019 Feb 5. pii: RA118.006173. doi: 10.1074/jbc.RA118.006173."
COMMENT     Sequence Label: pCEP4-HA-Flag-natT1R2 a22-end_I132S-S212N-D231G
FEATURES             Location/Qualifiers
     source          1..5654
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        86..231
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             274..1311
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPDRE
                     MGEAN"
     primer_bind     complement(1309..1328)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1353..1401
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     enhancer        2055..2434
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2435..2638
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2589..2609
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2635..2659
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     polyA_signal    5259..5393
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5283..5302)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5337..5356
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5536..5558)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gccctgcttc atccccgtgg cccgttgctc gcgtttgctg gcggtgtccc cggaagaaat
       61 atatttgcat gtctttagtt ctatgatgac acaaaccccg cccagcgtct tgtcattggc
      121 gaattcgaac acgcagatgc agtcggggcg gcgcggtccg aggtccactt cgcatattaa
      181 ggtgacgcgt gtggcctcga acaccgagcg accctgcagc gacccgctta acagcgtcaa
      241 cagcgtgccg cagatcccgg ggggcaatga gatatgaaaa agcctgaact caccgcgacg
      301 tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg
      361 gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg
      421 gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg
      481 gccgcgctcc cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat
      541 tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc
      601 gctgttctgc agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag
      661 acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat
      721 ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc
      781 gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc
      841 gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc
      901 cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc
      961 gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc
     1021 gagcggaggc atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt
     1081 ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg
     1141 cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc
     1201 gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga
     1261 aaccgacgcc ccagcactcg tccggatcgg gagatggggg aggctaactg aaacacggaa
     1321 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     1381 cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     1441 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     1501 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     1561 ccatagccac tggccccgtg ggttagggac ggggtccccc atggggaatg gtttatggtt
     1621 cgtgggggtt attattttgg gcgttgcgtg gggtcaggtc cacgactgga ctgagcagac
     1681 agacccatgg tttttggatg gcctgggcat ggaccgcatg tactggcgcg acacgaacac
     1741 cgggcgtctg tggctgccaa acacccccga cccccaaaaa ccaccgcgcg gatttctggc
     1801 gtgccaagct agtcgaccaa ttctcatgtt tgacagctta tcatcgcaga tccgggcaac
     1861 gttgttgcca ttgctgcagg cgcagaactg gtaggtatgg aagatctata cattgaatca
     1921 atattggcaa ttagccatat tagtcattgg ttatatagca taaatcaata ttggctattg
     1981 gccattgcat acgttgtatc tatatcataa tatgtacatt tatattggct catgtccaat
     2041 atgaccgcca tgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
     2101 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     2161 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     2221 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     2281 cttggcagta catcaagtgt atcatatgcc aagtccgccc cctattgacg tcaatgacgg
     2341 taaatggccc gcctggcatt atgcccagta catgacctta cgggactttc ctacttggca
     2401 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacaccaa
     2461 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     2521 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta ataaccccgc
     2581 cccgttgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg
     2641 tttagtgaac cgtcagatct ctagaagctg ggtaccagct gctagcgcca ccatgaagac
     2701 catcatcgcc ctgagctaca tcttctgcct ggtgttcgcc gactacaagg atgacgacga
     2761 cggcagcccc ggaggagctg ctgagaactc ggacttctac ctgcctgggg attacctcct
     2821 gggtggcctc ttctccctcc atgccaacat gaagggcatt gttcacctta acttcctgca
     2881 ggtgcccatg tgcaaggagt atgaagtgaa ggtgataggc tacaacctca tgcaggccat
     2941 gcgctttgcg gtggaggaga tcaacaatga cagcagcctg ctgcctggtg tgctgctggg
     3001 ctatgagatc gtggatgtgt gctacatctc caacaatgtc cagccggtgc tctacttcct
     3061 ggcacacgag gacaacctcc ttcccatcca agaggactac agtaactaca gttcccgtgt
     3121 ggtggctgtc attggccctg acaactccga gtctgtcatg actgtggcca acttcctctc
     3181 cctatttctc cttccacaga tcacctacag cgccatcagc gatgagctgc gagacaaggt
     3241 gcgcttcccg gctttgctgc gtaccacacc cagcgccgac caccacatcg aggccatggt
     3301 gcagcttatg ctgcacttcc gctggaactg gatcattgtg ctggtgagca acgacaccta
     3361 tggccgcgac aatggccagc tgcttggcga gcgcgtggcc cggcgcggca tctgcatcgc
     3421 cttccaggag acgctgccca cactgcagcc caaccagaac atgacgtcag aggagcgcca
     3481 gcgcctggtg accattgtgg acaagctgca gcagagcaca gcgcgcgtcg tggtcgtgtt
     3541 ctcgcccgac ctgaccctgt accacttctt caatgaggtg ctgcgccaga acttcactgg
     3601 cgccgtgtgg atcgcctccg agtcctgggc catcgacccg gtcctgcaca acctcacgga
     3661 gctgcgccac ttgggcacct tcctgggcat caccatccag agcgtgccca tcccgggctt
     3721 cagtgagttc cgcgagtggg gcccacaggc tgggccgcca cccctcagca ggaccagcca
     3781 gagctatacc tgcaaccagg agtgcgacaa ctgcctgaac gccaccttgt ccttcaacac
     3841 cattctcagg ctctctgggg agcgtgtcgt ctacagcgtg tactctgcgg tctatgctgt
     3901 ggcccatgcc ctgcacagcc tcctcggctg tgacaaaagc acctgcacca agagggtggt
     3961 ctacccctgg cagctgcttg aggagatctg gaaggtcaac ttcactctcc tggaccacca
     4021 aatcttcttc gacccgcaag gggacgtggc tctgcacttg gagattgtcc agtggcaatg
     4081 ggaccggagc cagaatccct tccagagcgt cgcctcctac taccccctgc agcgacagct
     4141 gaagaacatc caagacatct cctggcacac catcaacaac acgatcccta tgtccatgtg
     4201 ttccaagagg tgccagtcag ggcaaaagaa gaagcctgtg ggcatccacg tctgctgctt
     4261 cgagtgcatc gactgccttc ccggcacctt cctcaaccac actgaagatg aatatgaatg
     4321 ccaggcctgc ccgaataacg agtggtccta ccagagtgag acctcctgct tcaagcggca
     4381 gctggtcttc ctggaatggc atgaggcacc caccatcgct gtggccctgc tggccgccct
     4441 gggcttcctc agcaccctgg ccatcctggt gatattctgg aggcacttcc agacacccat
     4501 agttcgctcg gctgggggcc ccatgtgctt cctgatgctg acactgctgc tggtggcata
     4561 catggtggtc ccggtgtacg tggggccgcc caaggtctcc acctgcctct gccgccaggc
     4621 cctctttccc ctctgcttca caatctgcat ctcctgtatc gccgtgcgtt ctttccagat
     4681 cgtctgcgcc ttcaagatgg ccagccgctt cccacgcgcc tacagctact gggtccgcta
     4741 ccaggggccc tacgtctcta tggcatttat cacggtactc aaaatggtca ttgtggtaat
     4801 tggcatgctg gccacgggcc tcagtcccac cacccgtact gaccccgatg accccaagat
     4861 cacaattgtc tcctgtaacc ccaactaccg caacagcctg ctgttcaaca ccagcctgga
     4921 cctgctgctc tcagtggtgg gtttcagctt cgcctacatg ggcaaagagc tgcccaccaa
     4981 ctacaacgag gccaagttca tcaccctcag catgaccttc tatttcacct catccgtctc
     5041 cctctgcacc ttcatgtctg cctacagcgg ggtgctggtc accatcgtgg acctcttggt
     5101 cactgtgctc aacctcctgg ccatcagcct gggctacttc ggccccaagt gctacatgat
     5161 cctcttctac ccggagcgca acacgcccgc ctacttcaac agcatgatcc agggctacac
     5221 catgaggagg gactagtgac tcgaggccgg caaggccgga tccagacatg ataagataca
     5281 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     5341 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     5401 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     5461 agtaaaacct ctacaaatgt ggtatggctg attatgatcc ggctgcctcg cgcgtttcgg
     5521 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     5581 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     5641 gggcgcagcc atga
//
