LOCUS       sequence-287606-        5176 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 5176)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5176)
  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-m4046536
FEATURES             Location/Qualifiers
     source          1..5176
                     /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..886
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(931..949)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1300..2103
                     /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      2186..3168
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3246..3275
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3297..3884
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3896..3923)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3947..4138)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4139..4930,4931..4999))
                     /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     4762..4781
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5000..5102)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5103..5134)
                     /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 gaggttctca ccatcatcac catcattaaa caggcccctt
      901 ttcctttgtc gatatcatgt aattagttat gtcacgctta cattcacgcc ctcctcccac
      961 atccgctcta accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt
     1021 tttttaatag ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt
     1081 ctgtacaaag cggccgccga accactagtg cactgcagta cacggtttcc ttgaaatttt
     1141 tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg
     1201 gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa
     1261 ctgcacagaa caaaaacgtg caggaaacga agataaatca tgtcgaaagc tacatataag
     1321 gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa
     1381 aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta
     1441 gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat
     1501 ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta
     1561 ctcttcgagg acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg
     1621 ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca
     1681 ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt
     1741 ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt
     1801 actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac
     1861 atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat
     1921 gacaagggag atgcattggg tcaacagtat agaaccgtgg atgatgtggt ttctacagga
     1981 tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag
     2041 ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac
     2101 taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt
     2161 taattatatc agttattacc cgagtaacga agcatctgtg cttcattttg tagaacaaaa
     2221 atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca
     2281 gaaatgcaac gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa
     2341 caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca
     2401 gaacagaaat gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg
     2461 ttctacaaaa atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt
     2521 tttctccttt gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt
     2581 taaggttaga agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc
     2641 cacttcccgc gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc
     2701 atccccgatt atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata
     2761 gcgttgatga ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat
     2821 atactacgta taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt
     2881 tcttactaca atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag
     2941 gtcgagttta gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat
     3001 agcacagaga tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc
     3061 aatattttag tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcatca
     3121 gagcgctttt ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg
     3181 aacttcccga gcggccgcgt gttacaagct tgactccagc gtaactggac tgcaatcaac
     3241 tcactggctc accttcacgg gtgggccttt cttcggtaga aaatcaaagg atcttcttga
     3301 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3361 gtggtttgtt tgccggatca agagctacca actctttttc cgaggtaact ggcttcagca
     3421 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     3481 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3541 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3601 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3661 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3721 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3781 caggggtaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3841 atcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcag
     3901 aaaggcccac ccgaaggtga gccaggtgat tacatttggg ccctcatcag aggttttcac
     3961 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     4021 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     4081 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactt
     4141 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4201 ttgcctgact ccccgtcgtg tagataacta cgatgcggga gggcttacca tctggcccca
     4261 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4321 agccagccgg gagggccgag cgcagaagtg atcctgcaac tttatccgcc tccatccagt
     4381 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4441 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4501 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4561 ttagctcctt cggtcctccg atcgttgcca gaagtaagtt ggccgcagtg ttatcactca
     4621 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtgaga tgcttttctg
     4681 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4741 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4801 tcattggaaa acgttcttcg gggcgtaaac tctcaaggat cttaccgctg ttgagatcca
     4861 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4921 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4981 ggaaatgttg aatactcatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa
     5041 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat
     5101 caagtcaaaa gcctccggtc ggaggctttt gactttctgc tatggaggtc aggtatgatt
     5161 taaatggtca gttaga
//
