LOCUS       sequence-297673-        6256 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 6256)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6256)
  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-m4046411
FEATURES             Location/Qualifiers
     source          1..6256
                     /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"
     primer_bind     complement(2011..2029)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2380..3183
                     /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      3266..4248
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4326..4355
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4377..4964
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4976..5003)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5027..5218)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5219..6010,6011..6079))
                     /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     5842..5861
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6080..6182)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6183..6214)
                     /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 gaggttctaa aatcgaggaa ggtaagctag tcatttggat
      901 aaatggagat aagggttaca acggtttggc agaagtaggc aagaaatttg aaaaggacac
      961 aggtatcaag gtgacggttg aacatccaga caagttagaa gaaaaattcc ctcaagttgc
     1021 tgctactggt gatggtcccg acattatttt ctgggcccac gatcgttttg gtggatatgc
     1081 tcagtcaggt ttgctggctg agatcacccc agacaaggcc ttccaagata aattgtaccc
     1141 attcacttgg gatgctgtca gatacaacgg taagcttata gcatatccaa ttgctgttga
     1201 agccttatcg ttgatctaca ataaggactt gttacctaac ccaccaaaga catgggaaga
     1261 aattccagct ttggataaag agctaaaggc caagggaaaa agcgctttga tgtttaactt
     1321 acaagaacct tatttcacct ggccattaat cgctgctgac ggtggttacg catttaaata
     1381 cgaaaacggt aagtatgaca ttaaggatgt cggcgttgat aatgctggtg caaaggctgg
     1441 tttgaccttc ttggttgatt taataaagaa taaacacatg aacgctgaca ctgactactc
     1501 tatcgccgaa gccgctttca acaagggtga aactgctatg accataaatg gtccatgggc
     1561 ctggagtaac attgatactt ctaaagtaaa ctatggtgtc actgttttgc ctacattcaa
     1621 aggccaacca tccaagccct ttgtcggtgt tctatccgct ggcatcaatg ctgcttctcc
     1681 aaacaaggag ctagctaaag aatttttgga aaattacttg ttaactgacg aaggtttgga
     1741 ggccgttaac aaggacaagc cattgggtgc tgtcgctttg aaatcttacg aagaagaatt
     1801 ggctaaggat cccagaattg ccgctaccat ggagaacgcc caaaagggtg aaattatgcc
     1861 aaacatccca caaatgtcag ccttctggta cgcagttagg actgccgtga ttaatgctgc
     1921 ttccggtaga caaaccgtcg acgaagcatt aaaagatgct caaacgtaaa caggcccctt
     1981 ttcctttgtc gatatcatgt aattagttat gtcacgctta cattcacgcc ctcctcccac
     2041 atccgctcta accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt
     2101 tttttaatag ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt
     2161 ctgtacaaag cggccgccga accactagtg cactgcagta cacggtttcc ttgaaatttt
     2221 tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg
     2281 gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa
     2341 ctgcacagaa caaaaacgtg caggaaacga agataaatca tgtcgaaagc tacatataag
     2401 gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa
     2461 aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta
     2521 gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat
     2581 ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta
     2641 ctcttcgagg acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg
     2701 ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca
     2761 ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt
     2821 ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt
     2881 actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac
     2941 atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat
     3001 gacaagggag atgcattggg tcaacagtat agaaccgtgg atgatgtggt ttctacagga
     3061 tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag
     3121 ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac
     3181 taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt
     3241 taattatatc agttattacc cgagtaacga agcatctgtg cttcattttg tagaacaaaa
     3301 atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca
     3361 gaaatgcaac gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa
     3421 caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca
     3481 gaacagaaat gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg
     3541 ttctacaaaa atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt
     3601 tttctccttt gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt
     3661 taaggttaga agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc
     3721 cacttcccgc gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc
     3781 atccccgatt atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata
     3841 gcgttgatga ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat
     3901 atactacgta taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt
     3961 tcttactaca atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag
     4021 gtcgagttta gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat
     4081 agcacagaga tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc
     4141 aatattttag tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcatca
     4201 gagcgctttt ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg
     4261 aacttcccga gcggccgcgt gttacaagct tgactccagc gtaactggac tgcaatcaac
     4321 tcactggctc accttcacgg gtgggccttt cttcggtaga aaatcaaagg atcttcttga
     4381 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4441 gtggtttgtt tgccggatca agagctacca actctttttc cgaggtaact ggcttcagca
     4501 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     4561 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4621 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4681 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4741 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     4801 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4861 caggggtaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     4921 atcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcag
     4981 aaaggcccac ccgaaggtga gccaggtgat tacatttggg ccctcatcag aggttttcac
     5041 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     5101 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     5161 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactt
     5221 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     5281 ttgcctgact ccccgtcgtg tagataacta cgatgcggga gggcttacca tctggcccca
     5341 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5401 agccagccgg gagggccgag cgcagaagtg atcctgcaac tttatccgcc tccatccagt
     5461 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5521 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5581 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5641 ttagctcctt cggtcctccg atcgttgcca gaagtaagtt ggccgcagtg ttatcactca
     5701 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtgaga tgcttttctg
     5761 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5821 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5881 tcattggaaa acgttcttcg gggcgtaaac tctcaaggat cttaccgctg ttgagatcca
     5941 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6001 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6061 ggaaatgttg aatactcatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa
     6121 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat
     6181 caagtcaaaa gcctccggtc ggaggctttt gactttctgc tatggaggtc aggtatgatt
     6241 taaatggtca gttaga
//
