LOCUS       sequence-298361-        5134 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 5134)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5134)
  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-m4046649
FEATURES             Location/Qualifiers
     source          1..5134
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(889..907)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1258..2061
                     /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      2144..3126
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3204..3233
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3255..3842
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3854..3881)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3905..4096)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4097..4888,4889..4957))
                     /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     4720..4739
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4958..5060)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5061..5092)
                     /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 atgggtaatg caacagaagt gcctgccaat tcaactgtat tatctttctg
      481 tgcttttgct gtagatgctg ctaaagctta caaagattat ctagctagtg ggggacaacc
      541 aatcactaat tgtgttaaga tgttgtgtac acacactggt actggtcagg caataacagt
      601 tacaccggaa gccaatatgg atcaagaatc ctttggtggt gcatcgtgtt gtctgtactg
      661 ccgttgccac atagatcatc caaatcctaa aggattttgt gacttaaaag gtaagtatgt
      721 acaaatacct acaacttgtg ctaatgaccc tgtgggtttt acacttaaaa acacagtctg
      781 taccgtctgc ggtatgtgga aaggttatgg ctgtagttgt gatcaactcc gcgaacccat
      841 gctttaaaca ggcccctttt cctttgtcga tatcatgtaa ttagttatgt cacgcttaca
      901 ttcacgccct cctcccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt
      961 ctaggtccct atttattttt tttaatagtt atgttagtat taagaacgtt atttatattt
     1021 caaatttttc ttttttttct gtacaaagcg gccgccgaac cactagtgca ctgcagtaca
     1081 cggtttcctt gaaatttttt tgattcggta atctccgaac agaaggaaga acgaaggaag
     1141 gagcacagac ttagattggt atatatacgc atatgtagtg ttgaagaaac atgaaattgc
     1201 ccagtattct taacccaact gcacagaaca aaaacgtgca ggaaacgaag ataaatcatg
     1261 tcgaaagcta catataagga acgtgctgct actcatccta gtcctgttgc tgccaagcta
     1321 tttaatatca tgcacgaaaa gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc
     1381 aaggaattac tggagttagt tgaagcatta ggtcccaaaa tttgtttact aaaaacacat
     1441 gtggatatct tgactgattt ttccatggag ggcacagtta agccgctaaa ggcattatcc
     1501 gccaagtaca attttttact cttcgaggac agaaaatttg ctgacattgg taatacagtc
     1561 aaattgcagt actctgcggg tgtatacaga atagcagaat gggcagacat tacgaatgca
     1621 cacggtgtgg tgggcccagg tattgttagc ggtttgaagc aggcggcaga agaagtaaca
     1681 aaggaaccta gaggcctttt gatgttagca gaattgtcat gcaagggctc cctatctact
     1741 ggagaatata ctaagggtac tgttgacatt gcgaagagcg acaaagattt tgttatcggc
     1801 tttattgctc aaagagacat gggtggaaga gatgaaggtt acgattggtt gattatgaca
     1861 cccggtgtgg gtttagatga caagggagat gcattgggtc aacagtatag aaccgtggat
     1921 gatgtggttt ctacaggatc tgacattatt attgttggaa gaggactatt tgcaaaggga
     1981 agggatgcta aggtagaggg tgaacgttac agaaaagcag gctgggaagc atatttgaga
     2041 agatgcggcc agcaaaacta aaaaactgta ttataagtaa atgcatgtat actaaactca
     2101 caaattagag cttcaattta attatatcag ttattacccg agtaacgaag catctgtgct
     2161 tcattttgta gaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag
     2221 ctgcattttt acagaacaga aatgcaacgc gaaagcgcta ttttaccaac gaagaatctg
     2281 tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc taatttttca aacaaagaat
     2341 ctgagctgca tttttacaga acagaaatgc aacgcgagag cgctatttta ccaacaaaga
     2401 atctatactt cttttttgtt ctacaaaaat gcatcccgag agcgctattt ttctaacaaa
     2461 gcatcttaga ttactttttt tctcctttgt gcgctctata atgcagtctc ttgataactt
     2521 tttgcactgt aggtccgtta aggttagaag aaggctactt tggtgtctat tttctcttcc
     2581 ataaaaaaag cctgactcca cttcccgcgt ttactgatta ctagcgaagc tgcgggtgca
     2641 ttttttcaag ataaaggcat ccccgattat attctatacc gatgtggatt gcgcatactt
     2701 tgtgaacaga aagtgatagc gttgatgatt cttcattggt cagaaaatta tgaacggttt
     2761 cttctatttt gtctctatat actacgtata ggaaatgttt acattttcgt attgttttcg
     2821 attcactcta tgaatagttc ttactacaat ttttttgtct aaagagtaat actagagata
     2881 aacataaaaa atgtagaggt cgagtttaga tgcaagttca aggagcgaaa ggtggatggg
     2941 taggttatat agggatatag cacagagata tatagcaaag agatactttt gagcaatgtt
     3001 tgtggaagcg gtattcgcaa tattttagta gctcgttaca gtccggtgcg tttttggttt
     3061 tttgaaagtg cgtcatcaga gcgcttttgg ttttcaaaag cgctctgaag ttcctatact
     3121 ttctagctag agaataggaa cttcccgagc ggccgcgtgt tacaagcttg actccagcgt
     3181 aactggactg caatcaactc actggctcac cttcacgggt gggcctttct tcggtagaaa
     3241 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3301 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     3361 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     3421 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3481 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3541 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3601 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     3661 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3721 agcgcacgag ggagcttcca ggggtaaacg cctggtatct ttatagtcct gtcgggtttc
     3781 gccacctctg acttgagcat cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3841 aaaacgccag caacgcagaa aggcccaccc gaaggtgagc caggtgatta catttgggcc
     3901 ctcatcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat
     3961 cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga
     4021 gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt
     4081 tttcctgttt ggtcacttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4141 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atgcgggagg
     4201 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     4261 atttatcagc aataaaccag ccagccggga gggccgagcg cagaagtgat cctgcaactt
     4321 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4381 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4441 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4501 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgccaga agtaagttgg
     4561 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4621 ccgtgagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4681 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4741 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgtaaactc tcaaggatct
     4801 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4861 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4921 agggaataag ggcgacacgg aaatgttgaa tactcatttt agcttcctta gctcctgaaa
     4981 atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg
     5041 aacctcttac gtgccgatca agtcaaaagc ctccggtcgg aggcttttga ctttctgcta
     5101 tggaggtcag gtatgattta aatggtcagt taga
//
