LOCUS       sequence-292511-        5223 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 5223)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5223)
  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-m4046290
FEATURES             Location/Qualifiers
     source          1..5223
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        53..494
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    53..170
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     465..488
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             920..937
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             938..961
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1006..1024)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1375..2178
                     /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      2261..3243
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3321..3350
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3372..3959
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3971..3998)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4022..4213)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4214..5005,5006..5074))
                     /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     4837..4856
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5075..5177)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5178..5209)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 tcaggtatga tttaaatggt cagttagagc ggccgcttgc tatattgaag tacggattag
       61 aagccgccga gcgggcgaca gccctccgac ggaagactct cctccgtgcg tcctcgtctt
      121 caccggtcgc gttcctgaaa cgcagatgtg cctcgcgccg cactgctccg aacaataaag
      181 attctacaat actagctttt atggttatga agaggaaaaa ttggcagtaa cctggcccca
      241 caaaccttca aattaacgaa tcaaattaac aaccatagga tgataatgcg attagttttt
      301 tagccttatt tctggggtaa ttaatcagcg aagcgatgat ttttgatcta ttaacagata
      361 tataaatgga aaagctgcat aaccacttta actaatactt tcaacatttt cagtttgtat
      421 tacttcttat tcaaatgtca taaaagtatc aacaaaaaat tgttaatata cctctatact
      481 ttaacgtcaa ggagaaaaaa ctataatggg taatgcaaca gaagtgcctg ccaattcaac
      541 tgtattatct ttctgtgctt ttgctgtaga tgctgctaaa gcttacaaag attatctagc
      601 tagtggggga caaccaatca ctaattgtgt taagatgttg tgtacacaca ctggtactgg
      661 tcaggcaata acagttacac cggaagccaa tatggatcaa gaatcctttg gtggtgcatc
      721 gtgttgtctg tactgccgtt gccacataga tcatccaaat cctaaaggat tttgtgactt
      781 aaaaggtaag tatgtacaaa tacctacaac ttgtgctaat gaccctgtgg gttttacact
      841 taaaaacaca gtctgtaccg tctgcggtat gtggaaaggt tatggctgta gttgtgatca
      901 actccgcgaa cccatgcttc tcgtgccgag aggttcttgg agccatccgc agtttgaaaa
      961 ataaacaggc cccttttcct ttgtcgatat catgtaatta gttatgtcac gcttacattc
     1021 acgccctcct cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta
     1081 ggtccctatt tatttttttt aatagttatg ttagtattaa gaacgttatt tatatttcaa
     1141 atttttcttt tttttctgta caaagcggcc gccgaaccac tagtgcactg cagtacacgg
     1201 tttccttgaa atttttttga ttcggtaatc tccgaacaga aggaagaacg aaggaaggag
     1261 cacagactta gattggtata tatacgcata tgtagtgttg aagaaacatg aaattgccca
     1321 gtattcttaa cccaactgca cagaacaaaa acgtgcagga aacgaagata aatcatgtcg
     1381 aaagctacat ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt
     1441 aatatcatgc acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag
     1501 gaattactgg agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg
     1561 gatatcttga ctgatttttc catggagggc acagttaagc cgctaaaggc attatccgcc
     1621 aagtacaatt ttttactctt cgaggacaga aaatttgctg acattggtaa tacagtcaaa
     1681 ttgcagtact ctgcgggtgt atacagaata gcagaatggg cagacattac gaatgcacac
     1741 ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg cggcagaaga agtaacaaag
     1801 gaacctagag gccttttgat gttagcagaa ttgtcatgca agggctccct atctactgga
     1861 gaatatacta agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt
     1921 attgctcaaa gagacatggg tggaagagat gaaggttacg attggttgat tatgacaccc
     1981 ggtgtgggtt tagatgacaa gggagatgca ttgggtcaac agtatagaac cgtggatgat
     2041 gtggtttcta caggatctga cattattatt gttggaagag gactatttgc aaagggaagg
     2101 gatgctaagg tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga
     2161 tgcggccagc aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa
     2221 attagagctt caatttaatt atatcagtta ttacccgagt aacgaagcat ctgtgcttca
     2281 ttttgtagaa caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg
     2341 catttttaca gaacagaaat gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc
     2401 ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg
     2461 agctgcattt ttacagaaca gaaatgcaac gcgagagcgc tattttacca acaaagaatc
     2521 tatacttctt ttttgttcta caaaaatgca tcccgagagc gctatttttc taacaaagca
     2581 tcttagatta ctttttttct cctttgtgcg ctctataatg cagtctcttg ataacttttt
     2641 gcactgtagg tccgttaagg ttagaagaag gctactttgg tgtctatttt ctcttccata
     2701 aaaaaagcct gactccactt cccgcgttta ctgattacta gcgaagctgc gggtgcattt
     2761 tttcaagata aaggcatccc cgattatatt ctataccgat gtggattgcg catactttgt
     2821 gaacagaaag tgatagcgtt gatgattctt cattggtcag aaaattatga acggtttctt
     2881 ctattttgtc tctatatact acgtatagga aatgtttaca ttttcgtatt gttttcgatt
     2941 cactctatga atagttctta ctacaatttt tttgtctaaa gagtaatact agagataaac
     3001 ataaaaaatg tagaggtcga gtttagatgc aagttcaagg agcgaaaggt ggatgggtag
     3061 gttatatagg gatatagcac agagatatat agcaaagaga tacttttgag caatgtttgt
     3121 ggaagcggta ttcgcaatat tttagtagct cgttacagtc cggtgcgttt ttggtttttt
     3181 gaaagtgcgt catcagagcg cttttggttt tcaaaagcgc tctgaagttc ctatactttc
     3241 tagctagaga ataggaactt cccgagcggc cgcgtgttac aagcttgact ccagcgtaac
     3301 tggactgcaa tcaactcact ggctcacctt cacgggtggg cctttcttcg gtagaaaatc
     3361 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3421 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgagg
     3481 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     3541 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3601 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3661 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3721 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3781 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3841 gcacgaggga gcttccaggg gtaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3901 acctctgact tgagcatcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3961 acgccagcaa cgcagaaagg cccacccgaa ggtgagccag gtgattacat ttgggccctc
     4021 atcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     4081 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     4141 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     4201 cctgtttggt cacttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4261 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgatg cgggagggct
     4321 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4381 tatcagcaat aaaccagcca gccgggaggg ccgagcgcag aagtgatcct gcaactttat
     4441 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4501 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4561 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4621 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgccagaagt aagttggccg
     4681 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4741 tgagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4801 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4861 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg taaactctca aggatcttac
     4921 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4981 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5041 gaataagggc gacacggaaa tgttgaatac tcattttagc ttccttagct cctgaaaatc
     5101 tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac
     5161 ctcttacgtg ccgatcaagt caaaagcctc cggtcggagg cttttgactt tctgctatgg
     5221 agg
//
