LOCUS       sequence-287638-        5271 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp10.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5271)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5271)
  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"
COMMENT     Sequence Label: pGBW-m4046388
FEATURES             Location/Qualifiers
     source          1..5271
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..466
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    25..142
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     437..460
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             898..915
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             916..981
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1026..1044)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1395..2198
                     /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"
     rep_origin      2281..3263
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3341..3370
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3392..3979
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3991..4018)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4042..4233)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4234..5025,5026..5094))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4857..4876
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5095..5197)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5198..5229)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgctt gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg
       61 acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg
      121 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat
      181 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta
      241 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag
      301 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt
      361 taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta
      421 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aactataatg
      481 gcaggaaacg ctactgaagt tccagccaat tcaaccgtgt tgagcttttg tgctttcgcg
      541 gtagatgctg ccaaggctta caaagactat cttgcttccg gtgggcaacc cattacgaac
      601 tgcgtcaaga tgctgtgtac tcacacaggc accggtcagg ctataactgt taccccagag
      661 gccaatatgg accaagaatc gttcggtggt gcatcttgtt gtctctactg ccgttgtcat
      721 atcgatcacc caaaccctaa gggtttttgt gacttaaaag gtaagtacgt tcaaatccca
      781 actacctgtg ctaacgatcc agtcggtttc accttgaaga atacagtttg tactgtatgc
      841 ggtatgtgga aaggatacgg ttgttcttgc gatcaattga gagaacctat gttgcaactc
      901 gtgccgagag gttctgatta taaagatcat gatggtgatt ataaagatca tgatattgat
      961 tataaagatg atgatgataa ataaacaggc cccttttcct ttgtcgatat catgtaatta
     1021 gttatgtcac gcttacattc acgccctcct cccacatccg ctctaaccga aaaggaagga
     1081 gttagacaac ctgaagtcta ggtccctatt tatttttttt aatagttatg ttagtattaa
     1141 gaacgttatt tatatttcaa atttttcttt tttttctgta caaagcggcc gccgaaccac
     1201 tagtgcactg cagtacacgg tttccttgaa atttttttga ttcggtaatc tccgaacaga
     1261 aggaagaacg aaggaaggag cacagactta gattggtata tatacgcata tgtagtgttg
     1321 aagaaacatg aaattgccca gtattcttaa cccaactgca cagaacaaaa acgtgcagga
     1381 aacgaagata aatcatgtcg aaagctacat ataaggaacg tgctgctact catcctagtc
     1441 ctgttgctgc caagctattt aatatcatgc acgaaaagca aacaaacttg tgtgcttcat
     1501 tggatgttcg taccaccaag gaattactgg agttagttga agcattaggt cccaaaattt
     1561 gtttactaaa aacacatgtg gatatcttga ctgatttttc catggagggc acagttaagc
     1621 cgctaaaggc attatccgcc aagtacaatt ttttactctt cgaggacaga aaatttgctg
     1681 acattggtaa tacagtcaaa ttgcagtact ctgcgggtgt atacagaata gcagaatggg
     1741 cagacattac gaatgcacac ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg
     1801 cggcagaaga agtaacaaag gaacctagag gccttttgat gttagcagaa ttgtcatgca
     1861 agggctccct atctactgga gaatatacta agggtactgt tgacattgcg aagagcgaca
     1921 aagattttgt tatcggcttt attgctcaaa gagacatggg tggaagagat gaaggttacg
     1981 attggttgat tatgacaccc ggtgtgggtt tagatgacaa gggagatgca ttgggtcaac
     2041 agtatagaac cgtggatgat gtggtttcta caggatctga cattattatt gttggaagag
     2101 gactatttgc aaagggaagg gatgctaagg tagagggtga acgttacaga aaagcaggct
     2161 gggaagcata tttgagaaga tgcggccagc aaaactaaaa aactgtatta taagtaaatg
     2221 catgtatact aaactcacaa attagagctt caatttaatt atatcagtta ttacccgagt
     2281 aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc gctaattttt
     2341 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa agcgctattt
     2401 taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa
     2461 tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgagagcgc
     2521 tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca tcccgagagc
     2581 gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg ctctataatg
     2641 cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag gctactttgg
     2701 tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta ctgattacta
     2761 gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt ctataccgat
     2821 gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt cattggtcag
     2881 aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga aatgtttaca
     2941 ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt tttgtctaaa
     3001 gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc aagttcaagg
     3061 agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat agcaaagaga
     3121 tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct cgttacagtc
     3181 cggtgcgttt ttggtttttt gaaagtgcgt catcagagcg cttttggttt tcaaaagcgc
     3241 tctgaagttc ctatactttc tagctagaga ataggaactt cccgagcggc cgcgtgttac
     3301 aagcttgact ccagcgtaac tggactgcaa tcaactcact ggctcacctt cacgggtggg
     3361 cctttcttcg gtagaaaatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3421 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3481 taccaactct ttttccgagg taactggctt cagcagagcg cagataccaa atactgttct
     3541 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3601 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3661 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3721 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3781 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3841 cagggtcgga acaggagagc gcacgaggga gcttccaggg gtaaacgcct ggtatcttta
     3901 tagtcctgtc gggtttcgcc acctctgact tgagcatcga tttttgtgat gctcgtcagg
     3961 ggggcggagc ctatggaaaa acgccagcaa cgcagaaagg cccacccgaa ggtgagccag
     4021 gtgattacat ttgggccctc atcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
     4081 gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc
     4141 cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc
     4201 catgttaagg gcggtttttt cctgtttggt cacttaccaa tgcttaatca gtgaggcacc
     4261 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     4321 aactacgatg cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     4381 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccgggaggg ccgagcgcag
     4441 aagtgatcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     4501 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     4561 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4621 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4681 tgccagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4741 tcttactgtc atgccatccg tgagatgctt ttctgtgact ggtgagtact caaccaagtc
     4801 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4861 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4921 taaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     4981 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5041 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcattttagc
     5101 ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc
     5161 attatggtga aagttggaac ctcttacgtg ccgatcaagt caaaagcctc cggtcggagg
     5221 cttttgactt tctgctatgg aggtcaggta tgatttaaat ggtcagttag a
//
