LOCUS       sequence-297972-        6756 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6756)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6756)
  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-m4046177
FEATURES             Location/Qualifiers
     source          1..6756
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..466
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    25..142
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     437..460
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             1351..1368
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2511..2529)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2880..3683
                     /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      3766..4748
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4826..4855
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4877..5464
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5476..5503)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5527..5718)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5719..6510,6511..6579))
                     /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     6342..6361
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6580..6682)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6683..6714)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgctt gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg
       61 acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg
      121 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat
      181 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta
      241 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag
      301 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt
      361 taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta
      421 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aactataatg
      481 tctgctgtca aaagaactat taagggtacc catcactggt tgctgctaac aatcttaact
      541 tccttgttag tgcttgttca aagtacccag tggtcgttgt ttttcttctt gtacgagaac
      601 gcctttctcc cattcgcaat ggggataatt gctatgtcag cttttgccat gatgttcgtt
      661 aagcacaaac atgcgttttt gtgcttattc ttgttgcctt ctctcgctac ggtagcttat
      721 ttcaatatgg tctacatgcc agcttcctgg gttatgcgta tcatgacttg gttggacatg
      781 gttgatacca gcctttctgg tttcaagttg aaagattgtg tcatgtacgc atctgccgtt
      841 gtattactaa ttcttatgac tgctagaacc gtctatgacg atggcgcgag aagggtttgg
      901 acactaatga acgtgttgac tttagtttac aaggtctact atggaaatgc cttggaccaa
      961 gctatctcca tgtgggcttt aataatctca gtcacttcta actacagtgg tgttgttact
     1021 acagtgatgt ttttggctag aggtattgtc ttcatgtgtg ttgaatattg tcccattttc
     1081 tttatcaccg gtaacacttt acaatgcatt atgttggttt actgtttctt aggttacttt
     1141 tgtacctgtt atttcggcct attttgtttg ttgaatcgat actttagact cacattgggt
     1201 gtttacgatt atttggtcag cactcaagaa tttagatata tgaactccca aggtttgtta
     1261 ccaccaaaga actctattga tgccttcaag ttgaacatca agctgttagg tgtcggtgga
     1321 aaaccttgca tcaaggtagc caccgtccaa ctcgtgccga gaggttctaa aatcgaggaa
     1381 ggtaagctag tcatttggat aaatggagat aagggttaca acggtttggc agaagtaggc
     1441 aagaaatttg aaaaggacac aggtatcaag gtgacggttg aacatccaga caagttagaa
     1501 gaaaaattcc ctcaagttgc tgctactggt gatggtcccg acattatttt ctgggcccac
     1561 gatcgttttg gtggatatgc tcagtcaggt ttgctggctg agatcacccc agacaaggcc
     1621 ttccaagata aattgtaccc attcacttgg gatgctgtca gatacaacgg taagcttata
     1681 gcatatccaa ttgctgttga agccttatcg ttgatctaca ataaggactt gttacctaac
     1741 ccaccaaaga catgggaaga aattccagct ttggataaag agctaaaggc caagggaaaa
     1801 agcgctttga tgtttaactt acaagaacct tatttcacct ggccattaat cgctgctgac
     1861 ggtggttacg catttaaata cgaaaacggt aagtatgaca ttaaggatgt cggcgttgat
     1921 aatgctggtg caaaggctgg tttgaccttc ttggttgatt taataaagaa taaacacatg
     1981 aacgctgaca ctgactactc tatcgccgaa gccgctttca acaagggtga aactgctatg
     2041 accataaatg gtccatgggc ctggagtaac attgatactt ctaaagtaaa ctatggtgtc
     2101 actgttttgc ctacattcaa aggccaacca tccaagccct ttgtcggtgt tctatccgct
     2161 ggcatcaatg ctgcttctcc aaacaaggag ctagctaaag aatttttgga aaattacttg
     2221 ttaactgacg aaggtttgga ggccgttaac aaggacaagc cattgggtgc tgtcgctttg
     2281 aaatcttacg aagaagaatt ggctaaggat cccagaattg ccgctaccat ggagaacgcc
     2341 caaaagggtg aaattatgcc aaacatccca caaatgtcag ccttctggta cgcagttagg
     2401 actgccgtga ttaatgctgc ttccggtaga caaaccgtcg acgaagcatt aaaagatgct
     2461 caaacgtaaa caggcccctt ttcctttgtc gatatcatgt aattagttat gtcacgctta
     2521 cattcacgcc ctcctcccac atccgctcta accgaaaagg aaggagttag acaacctgaa
     2581 gtctaggtcc ctatttattt tttttaatag ttatgttagt attaagaacg ttatttatat
     2641 ttcaaatttt tctttttttt ctgtacaaag cggccgccga accactagtg cactgcagta
     2701 cacggtttcc ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga
     2761 aggagcacag acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt
     2821 gcccagtatt cttaacccaa ctgcacagaa caaaaacgtg caggaaacga agataaatca
     2881 tgtcgaaagc tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc
     2941 tatttaatat catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca
     3001 ccaaggaatt actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac
     3061 atgtggatat cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat
     3121 ccgccaagta caatttttta ctcttcgagg acagaaaatt tgctgacatt ggtaatacag
     3181 tcaaattgca gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg
     3241 cacacggtgt ggtgggccca ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa
     3301 caaaggaacc tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctatcta
     3361 ctggagaata tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg
     3421 gctttattgc tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga
     3481 cacccggtgt gggtttagat gacaagggag atgcattggg tcaacagtat agaaccgtgg
     3541 atgatgtggt ttctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg
     3601 gaagggatgc taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga
     3661 gaagatgcgg ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact
     3721 cacaaattag agcttcaatt taattatatc agttattacc cgagtaacga agcatctgtg
     3781 cttcattttg tagaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg
     3841 agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc tattttacca acgaagaatc
     3901 tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc gctaattttt caaacaaaga
     3961 atctgagctg catttttaca gaacagaaat gcaacgcgag agcgctattt taccaacaaa
     4021 gaatctatac ttcttttttg ttctacaaaa atgcatcccg agagcgctat ttttctaaca
     4081 aagcatctta gattactttt tttctccttt gtgcgctcta taatgcagtc tcttgataac
     4141 tttttgcact gtaggtccgt taaggttaga agaaggctac tttggtgtct attttctctt
     4201 ccataaaaaa agcctgactc cacttcccgc gtttactgat tactagcgaa gctgcgggtg
     4261 cattttttca agataaaggc atccccgatt atattctata ccgatgtgga ttgcgcatac
     4321 tttgtgaaca gaaagtgata gcgttgatga ttcttcattg gtcagaaaat tatgaacggt
     4381 ttcttctatt ttgtctctat atactacgta taggaaatgt ttacattttc gtattgtttt
     4441 cgattcactc tatgaatagt tcttactaca atttttttgt ctaaagagta atactagaga
     4501 taaacataaa aaatgtagag gtcgagttta gatgcaagtt caaggagcga aaggtggatg
     4561 ggtaggttat atagggatat agcacagaga tatatagcaa agagatactt ttgagcaatg
     4621 tttgtggaag cggtattcgc aatattttag tagctcgtta cagtccggtg cgtttttggt
     4681 tttttgaaag tgcgtcatca gagcgctttt ggttttcaaa agcgctctga agttcctata
     4741 ctttctagct agagaatagg aacttcccga gcggccgcgt gttacaagct tgactccagc
     4801 gtaactggac tgcaatcaac tcactggctc accttcacgg gtgggccttt cttcggtaga
     4861 aaatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4921 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4981 cgaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5041 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     5101 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     5161 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     5221 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     5281 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     5341 agagcgcacg agggagcttc caggggtaaa cgcctggtat ctttatagtc ctgtcgggtt
     5401 tcgccacctc tgacttgagc atcgattttt gtgatgctcg tcaggggggc ggagcctatg
     5461 gaaaaacgcc agcaacgcag aaaggcccac ccgaaggtga gccaggtgat tacatttggg
     5521 ccctcatcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc
     5581 atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt
     5641 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt
     5701 tttttcctgt ttggtcactt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5761 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatgcggga
     5821 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5881 agatttatca gcaataaacc agccagccgg gagggccgag cgcagaagtg atcctgcaac
     5941 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6001 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6061 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6121 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgcca gaagtaagtt
     6181 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6241 atccgtgaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6301 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6361 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgtaaac tctcaaggat
     6421 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6481 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6541 aaagggaata agggcgacac ggaaatgttg aatactcatt ttagcttcct tagctcctga
     6601 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt
     6661 ggaacctctt acgtgccgat caagtcaaaa gcctccggtc ggaggctttt gactttctgc
     6721 tatggaggtc aggtatgatt taaatggtca gttaga
//
