LOCUS       sequence-298902-        4986 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 ORF6 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4986)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4986)
  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-m4046589
FEATURES             Location/Qualifiers
     source          1..4986
                     /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             661..678
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             679..696
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(741..759)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1110..1913
                     /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      1996..2978
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3056..3085
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3107..3694
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3706..3733)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3757..3948)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3949..4740,4741..4809))
                     /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     4572..4591
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4810..4912)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4913..4944)
                     /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 tttcacctgg tcgacttcca agttacaatc gcagagattt tacttataat catgaggact
      541 ttcaaggtgt ctatttggaa tttggattac attatcaact tgatcattaa aaacttgtca
      601 aagagtttga ccgaaaataa gtattcccag ttagatgaag aacaacctat ggaaatagac
      661 ctcgtgccga gaggttctca ccatcatcac catcattaaa caggcccctt ttcctttgtc
      721 gatatcatgt aattagttat gtcacgctta cattcacgcc ctcctcccac atccgctcta
      781 accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt tttttaatag
      841 ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacaaag
      901 cggccgccga accactagtg cactgcagta cacggtttcc ttgaaatttt tttgattcgg
      961 taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg gtatatatac
     1021 gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa ctgcacagaa
     1081 caaaaacgtg caggaaacga agataaatca tgtcgaaagc tacatataag gaacgtgctg
     1141 ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa aagcaaacaa
     1201 acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta gttgaagcat
     1261 taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat ttttccatgg
     1321 agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta ctcttcgagg
     1381 acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg ggtgtataca
     1441 gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca ggtattgtta
     1501 gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt ttgatgttag
     1561 cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt actgttgaca
     1621 ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac atgggtggaa
     1681 gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat gacaagggag
     1741 atgcattggg tcaacagtat agaaccgtgg atgatgtggt ttctacagga tctgacatta
     1801 ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt
     1861 acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac taaaaaactg
     1921 tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt taattatatc
     1981 agttattacc cgagtaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg
     2041 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     2101 gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca
     2161 acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat
     2221 gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg ttctacaaaa
     2281 atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt tttctccttt
     2341 gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt taaggttaga
     2401 agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc cacttcccgc
     2461 gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc atccccgatt
     2521 atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga
     2581 ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat atactacgta
     2641 taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt tcttactaca
     2701 atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag gtcgagttta
     2761 gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat agcacagaga
     2821 tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc aatattttag
     2881 tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcatca gagcgctttt
     2941 ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg aacttcccga
     3001 gcggccgcgt gttacaagct tgactccagc gtaactggac tgcaatcaac tcactggctc
     3061 accttcacgg gtgggccttt cttcggtaga aaatcaaagg atcttcttga gatccttttt
     3121 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3181 tgccggatca agagctacca actctttttc cgaggtaact ggcttcagca gagcgcagat
     3241 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3301 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3361 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3421 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3481 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3541 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc caggggtaaa
     3601 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc atcgattttt
     3661 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcag aaaggcccac
     3721 ccgaaggtga gccaggtgat tacatttggg ccctcatcag aggttttcac cgtcatcacc
     3781 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     3841 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     3901 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactt accaatgctt
     3961 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4021 ccccgtcgtg tagataacta cgatgcggga gggcttacca tctggcccca gtgctgcaat
     4081 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4141 gagggccgag cgcagaagtg atcctgcaac tttatccgcc tccatccagt ctattaattg
     4201 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4261 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4321 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4381 cggtcctccg atcgttgcca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4441 agcactgcat aattctctta ctgtcatgcc atccgtgaga tgcttttctg tgactggtga
     4501 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4561 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4621 acgttcttcg gggcgtaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4681 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4741 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4801 aatactcatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
     4861 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat caagtcaaaa
     4921 gcctccggtc ggaggctttt gactttctgc tatggaggtc aggtatgatt taaatggtca
     4981 gttaga
//
