LOCUS       sequence-297726-        6246 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 ORF8 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6246)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6246)
  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-m4046296
FEATURES             Location/Qualifiers
     source          1..6246
                     /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             841..858
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2001..2019)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2370..3173
                     /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      3256..4238
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4316..4345
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4367..4954
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4966..4993)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5017..5208)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5209..6000,6001..6069))
                     /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     5832..5851
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6070..6172)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6173..6204)
                     /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 aaatttcttg ttttcttagg aatcatcaca actgtagctg catttcacca agaatgtagt
      541 ttacagtcat gtactcaaca tcaaccatat gtagttgatg acccgtgtcc tattcacttc
      601 tattctaaat ggtatattag agtaggagct agaaaatcag cacctttaat tgaattgtgc
      661 gtggatgagg ctggttctaa atcacccatt cagtacatcg atatcggtaa ttatacagtt
      721 tcctgtttac cttttacaat taattgccag gaacctaaat tgggtagtct tgtagtgcgt
      781 tgttcgttct atgaagactt tttagagtat catgacgttc gtgttgtttt agatttcatc
      841 ctcgtgccga gaggttctaa aatcgaggaa ggtaagctag tcatttggat aaatggagat
      901 aagggttaca acggtttggc agaagtaggc aagaaatttg aaaaggacac aggtatcaag
      961 gtgacggttg aacatccaga caagttagaa gaaaaattcc ctcaagttgc tgctactggt
     1021 gatggtcccg acattatttt ctgggcccac gatcgttttg gtggatatgc tcagtcaggt
     1081 ttgctggctg agatcacccc agacaaggcc ttccaagata aattgtaccc attcacttgg
     1141 gatgctgtca gatacaacgg taagcttata gcatatccaa ttgctgttga agccttatcg
     1201 ttgatctaca ataaggactt gttacctaac ccaccaaaga catgggaaga aattccagct
     1261 ttggataaag agctaaaggc caagggaaaa agcgctttga tgtttaactt acaagaacct
     1321 tatttcacct ggccattaat cgctgctgac ggtggttacg catttaaata cgaaaacggt
     1381 aagtatgaca ttaaggatgt cggcgttgat aatgctggtg caaaggctgg tttgaccttc
     1441 ttggttgatt taataaagaa taaacacatg aacgctgaca ctgactactc tatcgccgaa
     1501 gccgctttca acaagggtga aactgctatg accataaatg gtccatgggc ctggagtaac
     1561 attgatactt ctaaagtaaa ctatggtgtc actgttttgc ctacattcaa aggccaacca
     1621 tccaagccct ttgtcggtgt tctatccgct ggcatcaatg ctgcttctcc aaacaaggag
     1681 ctagctaaag aatttttgga aaattacttg ttaactgacg aaggtttgga ggccgttaac
     1741 aaggacaagc cattgggtgc tgtcgctttg aaatcttacg aagaagaatt ggctaaggat
     1801 cccagaattg ccgctaccat ggagaacgcc caaaagggtg aaattatgcc aaacatccca
     1861 caaatgtcag ccttctggta cgcagttagg actgccgtga ttaatgctgc ttccggtaga
     1921 caaaccgtcg acgaagcatt aaaagatgct caaacgtaaa caggcccctt ttcctttgtc
     1981 gatatcatgt aattagttat gtcacgctta cattcacgcc ctcctcccac atccgctcta
     2041 accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt tttttaatag
     2101 ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacaaag
     2161 cggccgccga accactagtg cactgcagta cacggtttcc ttgaaatttt tttgattcgg
     2221 taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg gtatatatac
     2281 gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa ctgcacagaa
     2341 caaaaacgtg caggaaacga agataaatca tgtcgaaagc tacatataag gaacgtgctg
     2401 ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa aagcaaacaa
     2461 acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta gttgaagcat
     2521 taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat ttttccatgg
     2581 agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta ctcttcgagg
     2641 acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg ggtgtataca
     2701 gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca ggtattgtta
     2761 gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt ttgatgttag
     2821 cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt actgttgaca
     2881 ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac atgggtggaa
     2941 gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat gacaagggag
     3001 atgcattggg tcaacagtat agaaccgtgg atgatgtggt ttctacagga tctgacatta
     3061 ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt
     3121 acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac taaaaaactg
     3181 tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt taattatatc
     3241 agttattacc cgagtaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg
     3301 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     3361 gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca
     3421 acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat
     3481 gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg ttctacaaaa
     3541 atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt tttctccttt
     3601 gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt taaggttaga
     3661 agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc cacttcccgc
     3721 gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc atccccgatt
     3781 atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga
     3841 ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat atactacgta
     3901 taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt tcttactaca
     3961 atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag gtcgagttta
     4021 gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat agcacagaga
     4081 tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc aatattttag
     4141 tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcatca gagcgctttt
     4201 ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg aacttcccga
     4261 gcggccgcgt gttacaagct tgactccagc gtaactggac tgcaatcaac tcactggctc
     4321 accttcacgg gtgggccttt cttcggtaga aaatcaaagg atcttcttga gatccttttt
     4381 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4441 tgccggatca agagctacca actctttttc cgaggtaact ggcttcagca gagcgcagat
     4501 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4561 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4621 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4681 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4741 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4801 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc caggggtaaa
     4861 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc atcgattttt
     4921 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcag aaaggcccac
     4981 ccgaaggtga gccaggtgat tacatttggg ccctcatcag aggttttcac cgtcatcacc
     5041 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     5101 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     5161 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactt accaatgctt
     5221 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5281 ccccgtcgtg tagataacta cgatgcggga gggcttacca tctggcccca gtgctgcaat
     5341 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5401 gagggccgag cgcagaagtg atcctgcaac tttatccgcc tccatccagt ctattaattg
     5461 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5521 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5581 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5641 cggtcctccg atcgttgcca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5701 agcactgcat aattctctta ctgtcatgcc atccgtgaga tgcttttctg tgactggtga
     5761 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5821 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5881 acgttcttcg gggcgtaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5941 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6001 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6061 aatactcatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
     6121 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat caagtcaaaa
     6181 gcctccggtc ggaggctttt gactttctgc tatggaggtc aggtatgatt taaatggtca
     6241 gttaga
//
