LOCUS       sequence-287583-        5182 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 5182)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5182)
  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-m4046197
FEATURES             Location/Qualifiers
     source          1..5182
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             851..868
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             869..892
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(937..955)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1306..2109
                     /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      2192..3174
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3252..3281
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3303..3890
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3902..3929)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3953..4144)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4145..4936,4937..5005))
                     /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     4768..4787
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5006..5108)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5109..5140)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat
       61 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
      121 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
      181 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg
      241 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
      301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      421 taattacaaa atggcaggaa acgctactga agttccagcc aattcaaccg tgttgagctt
      481 ttgtgctttc gcggtagatg ctgccaaggc ttacaaagac tatcttgctt ccggtgggca
      541 acccattacg aactgcgtca agatgctgtg tactcacaca ggcaccggtc aggctataac
      601 tgttacccca gaggccaata tggaccaaga atcgttcggt ggtgcatctt gttgtctcta
      661 ctgccgttgt catatcgatc acccaaaccc taagggtttt tgtgacttaa aaggtaagta
      721 cgttcaaatc ccaactacct gtgctaacga tccagtcggt ttcaccttga agaatacagt
      781 ttgtactgta tgcggtatgt ggaaaggata cggttgttct tgcgatcaat tgagagaacc
      841 tatgttgcaa ctcgtgccga gaggttcttg gagccatccg cagtttgaaa aataaacagg
      901 ccccttttcc tttgtcgata tcatgtaatt agttatgtca cgcttacatt cacgccctcc
      961 tcccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat
     1021 ttattttttt taatagttat gttagtatta agaacgttat ttatatttca aatttttctt
     1081 ttttttctgt acaaagcggc cgccgaacca ctagtgcact gcagtacacg gtttccttga
     1141 aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt
     1201 agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta
     1261 acccaactgc acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca
     1321 tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg
     1381 cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg
     1441 gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg
     1501 actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat
     1561 tttttactct tcgaggacag aaaatttgct gacattggta atacagtcaa attgcagtac
     1621 tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg
     1681 ggcccaggta ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga
     1741 ggccttttga tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact
     1801 aagggtactg ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa
     1861 agagacatgg gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt
     1921 ttagatgaca agggagatgc attgggtcaa cagtatagaa ccgtggatga tgtggtttct
     1981 acaggatctg acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag
     2041 gtagagggtg aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag
     2101 caaaactaaa aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct
     2161 tcaatttaat tatatcagtt attacccgag taacgaagca tctgtgcttc attttgtaga
     2221 acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac
     2281 agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt
     2341 gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     2401 tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct
     2461 tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt
     2521 actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag
     2581 gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc
     2641 tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat
     2701 aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa
     2761 gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt
     2821 ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg
     2881 aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat
     2941 gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag
     3001 ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt
     3061 attcgcaata ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg
     3121 tcatcagagc gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag
     3181 aataggaact tcccgagcgg ccgcgtgtta caagcttgac tccagcgtaa ctggactgca
     3241 atcaactcac tggctcacct tcacgggtgg gcctttcttc ggtagaaaat caaaggatct
     3301 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3361 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgag gtaactggct
     3421 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     3481 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3541 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3601 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3661 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3721 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3781 agcttccagg ggtaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3841 ttgagcatcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3901 acgcagaaag gcccacccga aggtgagcca ggtgattaca tttgggccct catcagaggt
     3961 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt
     4021 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa
     4081 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg
     4141 tcacttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4201 ccatagttgc ctgactcccc gtcgtgtaga taactacgat gcgggagggc ttaccatctg
     4261 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4321 taaaccagcc agccgggagg gccgagcgca gaagtgatcc tgcaacttta tccgcctcca
     4381 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4441 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4501 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4561 aagcggttag ctccttcggt cctccgatcg ttgccagaag taagttggcc gcagtgttat
     4621 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtgagatgct
     4681 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4741 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     4801 tgctcatcat tggaaaacgt tcttcggggc gtaaactctc aaggatctta ccgctgttga
     4861 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     4921 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     4981 cgacacggaa atgttgaata ctcattttag cttccttagc tcctgaaaat ctcgataact
     5041 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt
     5101 gccgatcaag tcaaaagcct ccggtcggag gcttttgact ttctgctatg gaggtcaggt
     5161 atgatttaaa tggtcagtta ga
//
