LOCUS       sequence-321534-        6565 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of proteins with a C-terminal thermostable GFP (TGP) tag 
            in Saccharomyces cerevisiae.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6565)
  AUTHORS   Cai H, Yao H, Li T, Hutter CAJ, Li Y, Tang Y, Seeger MA, Li D
  TITLE     An improved fluorescent tag and its nanobodies for membrane protein 
            expression, stability assay, and purification.
  JOURNAL   Commun Biol. 2020 Dec 10;3(1):753. doi: 10.1038/s42003-020-01478-z.
  PUBMED    33303987
REFERENCE   2  (bases 1 to 6565)
  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: "Commun 
            Biol."; date: "2020-12-10"; volume: "3(1):753. doi"; pages: " 
            10.1038/s42003-020-01478-z"
COMMENT     Sequence Label: pYTSG
FEATURES             Location/Qualifiers
     source          1..6565
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(151..366)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      967..1844
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      complement(1855..2368)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2050..2071)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2262..2281
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2390..2831
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    2390..2507
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     2802..2825
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     2863..2882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2863..2881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2940..2963
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3657..3677
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     terminator      3702..3949
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3718..3736)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4026..4043)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4197..4785)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4277..4296)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4956..5747,5748..5816))
                     /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     5579..5598
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             complement(join(5912..6565,1..150))
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
ORIGIN
        1 tgcttcaact aactccagta attccttggt ggtacgaaca tccaatgaag cacacaagtt
       61 tgtttgcttt tcgtgcatga tattaaatag cttggcagca acaggactag gatgagtagc
      121 agcacgttcc ttatatgtag ctttcgacat gatttatctt cgtttcctgc aggtttttgt
      181 tctgtgcagt tgggttaaga atactgggca atttcatgtt tcttcaacac tacatatgcg
      241 tatatatacc aatctaagtc tgtgctcctt ccttcgttct tccttctgtt cggagattac
      301 cgaatcaaaa aaatttcaag gaaaccgaaa tcaaaaaaaa gaataaaaaa aaaatgatga
      361 attgaaaagc tagcttatcg atgataagct gtcaaacatg agaattaatt ccacggacta
      421 tagactatac ctagtatact ccgtctactg tacgatacac ttccgctcag gtccttgtcc
      481 tttaacgagg ccttaccact cttttgttac tctattgatc cagctcagca aaggcagtgt
      541 gatctaagat tctatcttcg cgatgtagta aaactagcta gaccgagaaa gagactagaa
      601 atgcaaaagg cacttctaca atggctgcca tcattattat ccgatgtgac gctgcagctt
      661 ctcaatgata ttcgaatacg ctttgaggag atacagccta atatccgaca aactgtttta
      721 cagatttacg atcgtacttg ttacccatca ttgaattttg aacatccgaa cctgggagtt
      781 ttccctgaaa cagatagtat atttgaacct gtataataat atatagtcta gcgctttacg
      841 gaagacaatg tatgtatttc ggttcctgga gaaactattg catctattgc ataggtaatc
      901 ttgcacgtcg catccccggt tcattttctg cgtttccatc ttgcacttca atagcatatc
      961 tttgttaacg aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta
     1021 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg
     1081 ctattttacc aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag
     1141 cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga
     1201 gagcgctatt ttaccaacaa agaatctata cttctttttt gttctacaaa aatgcatccc
     1261 gagagcgcta tttttctaac aaagcatctt agattacttt ttttctcctt tgtgcgctct
     1321 ataatgcagt ctcttgataa ctttttgcac tgtaggtccg ttaaggttag aagaaggcta
     1381 ctttggtgtc tattttctct tccataaaaa aagcctgact ccacttcccg cgtttactga
     1441 ttactagcga agctgcgggt gcattttttc aagataaagg catccccgat tatattctat
     1501 accgatgtgg attgcgcata ctttgtgaac agaaagtgat agcgttgatg attcttcatt
     1561 ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta tatactacgt ataggaaatg
     1621 tttacatttt cgtattgttt tcgattcact ctatgaatag ttcttactac aatttttttg
     1681 tctaaagagt aatactagag ataaacataa aaaatgtaga ggtcgagttt agatgcaagt
     1741 tcaaggagcg aaaggtggat gggtaggtta tatagggata tagcacagag atatatagca
     1801 aagagatact tttgagcaat gtttgtggaa gcggtattcg caatgggaag ctccaccccg
     1861 gttgataatc agaaaagccc caaaaacagg aagattgtat aagcaaatat ttaaattgta
     1921 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     1981 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     2041 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     2101 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     2161 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     2221 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     2281 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     2341 gcgcttaatg cgccgctaca gggcgcgtgg ggatcgatcc actagtccac ggattagaag
     2401 ccgccgagcg ggtgacagcc ctccgaagga agactctcct ccgtgcgtcc tcgtcttcac
     2461 cggtcgcgtt cctgaaacgc agatgtgcct cgcgccgcac tgctccgaac aataaagatt
     2521 ctacaatact agcttttatg gttatgaaga ggaaaaattg gcagtaacct ggccccacaa
     2581 accttcaaat gaacgaatca aattaacaac cataggatga taatgcgatt agttttttag
     2641 ccttatttct ggggtaatta atcagcgaag cgatgatttt tgatctatta acagatatat
     2701 aaatgcaaaa actgcataac cactttaact aatactttca acattttcgg tttgtattac
     2761 ttcttattca aatgtaataa aagtatcaac aaaaaattgt taatatacct ctatacttta
     2821 acgtcaagga gaaaaaaccc cggatcggac tactagcagc tgtaatacga ctcactatag
     2881 ggaatattta aaaaatgtct ggatccaagc ttagcggccg cgaattcctc gagggtaccc
     2941 tggaagtgct gtttcagggc ccgggcggta gtggaggttc cgcgagtgtc attaagccgg
     3001 agatgaaaat taagttgcgt atggaggggg ccgtgaatgg gcacaaattc gttattgaag
     3061 gggaggggat tggaaagccg tatgagggta cgcaaacctt ggacttaaca gtagaagaag
     3121 gggcaccatt gcctttttct tatgatattt tgactccggc gttccaatac gggaaccgtg
     3181 ctttcacaaa gtatcccgaa gatatccctg actatttcaa gcaagcattt cccgagggtt
     3241 acagttggga gagaagtatg acctatgagg atcaaggcat atgtattgct acctctgaca
     3301 ttactatgga gggagactgc ttcttctatg agattcgttt cgatggaaca aactttcctc
     3361 ctaacgggcc agtgatgcag aaaaagacgt tgaagtggga accgtccaca gaaaaaatgt
     3421 atgtcgagga cggcgttcta aagggagacg tggaaatggc cttgctattg gaagggggtg
     3481 gacactatcg ttgtgatttc aaaacgacct acaaggccaa aaaagatgtg agactaccgg
     3541 acgcccacga ggttgaccat cgtattgaaa ttttaagtca cgacaaggac tataataaag
     3601 taagactgta cgaacacgcg gaagcacgtt acagcggagg tggaagtggt ggtggccatc
     3661 accatcacca tcaccattag cggccgctct agagggccgc atcatgtaat tagttatgtc
     3721 acgcttacat tcacgccctc cccccacatc cgctctaacc gaaaaggaag gagttagaca
     3781 acctgaagtc taggtcccta tttatttttt tatagttatg ttagtattaa gaacgttatt
     3841 tatatttcaa atttttcttt tttttctgta cagacgcgtg tacgcatgta acattatact
     3901 gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct ttaatttgca agctgcggcc
     3961 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     4021 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4081 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4141 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4201 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4261 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4321 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4381 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4441 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4501 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4561 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4621 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4681 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4741 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4801 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4861 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4921 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4981 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     5041 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5101 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5161 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5221 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5281 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5341 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5401 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5461 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5521 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5581 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5641 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5701 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5761 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5821 ttcctttttc aatgggtaat aactgatata attaaattga agctctaatt tgtgagttta
     5881 gtatacatgc atttacttat aatacagttt tttagttttg ctggccgcat cttctcaaat
     5941 atgcttccca gcctgctttt ctgtaacgtt caccctctac cttagcatcc cttccctttg
     6001 caaatagtcc tcttccaaca ataataatgt cagatcctgt agagaccaca tcatccacgg
     6061 ttctatactg ttgacccaat gcgtctccct tgtcatctaa acccacaccg ggtgtcataa
     6121 tcaaccaatc gtaaccttca tctcttccac ccatgtctct ttgagcaata aagccgataa
     6181 caaaatcttt gtcgctcttc gcaatgtcaa cagtaccctt agtatattct ccagtagata
     6241 gggagccctt gcatgacaat tctgctaaca tcaaaaggcc tctaggttcc tttgttactt
     6301 cttctgccgc ctgcttcaaa ccgctaacaa tacctgggcc caccacaccg tgtgcattcg
     6361 taatgtctgc ccattctgct attctgtata cacccgcaga gtactgcaat ttgactgtat
     6421 taccaatgtc agcaaatttt ctgtcttcga agagtaaaaa attgtacttg gcggataatg
     6481 cctttagcgg cttaactgtg ccctccatgg aaaaatcagt caagatatcc acatgtgttt
     6541 ttagtaaaca aattttggga cctaa
//
