LOCUS       sequence-298841-        6306 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6306)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6306)
  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-m4046337
FEATURES             Location/Qualifiers
     source          1..6306
                     /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             1981..1998
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1999..2016
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2061..2079)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2430..3233
                     /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      3316..4298
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4376..4405
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4427..5014
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5026..5053)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5077..5268)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5269..6060,6061..6129))
                     /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     5892..5911
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6130..6232)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6233..6264)
                     /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 aagattgtca acaattggct taaacagttg ataaaggtta ctttagtatt tttgttcgtt
      541 gcagccatct tctatctaat tacacctgtc cacgtgatga gtaagcatac cgacttttcc
      601 tcagagatca ttggttacaa agctatcgat ggcggagtta ctagagatat tgcttcgacg
      661 gacacctgtt tcgctaacaa gcacgcggac tttgatactt ggttctctca aaggggtggt
      721 tcctacacaa acgacaaggc ctgccccttg attgcagctg ttatcactag agaagttggt
      781 ttcgtcgtgc caggtttacc agggactata ctgcgtacca ccaatggtga ttttttgcat
      841 ttcttaccaa gagtcttctc tgctgttggc aatatttgtt atacccctag caaattgatc
      901 gaatacactg attttgccac ctctgcttgt gtattggctg ctgaatgcac tattttcaag
      961 gatgcaagtg gtaagccagt tccatactgt tatgacacaa acgtcctcga aggttctgtt
     1021 gcttacgaat ccttgcgacc ggacactaga tatgtgttaa tggatggatc aatcattcaa
     1081 tttccaaaca cataccttga aggttctgtt cgggttgtca ccaccttcga ctccgagtac
     1141 tgtcgccacg gtacttgtga aagatcagaa gccggtgtct gtgtttctac cagcggtaga
     1201 tgggtgttga ataacgatta ttaccgttct cttccaggtg tcttctgcgg tgtcgatgct
     1261 gttaatctat tgactaacat gtttacccct ttgatccaac caataggtgc gctggacatt
     1321 tccgcatcta ttgtcgccgg tggaatcgtt gctatcgttg ttacttgttt ggcttactac
     1381 ttcatgagat ttagaagagc ctttggggag tattcgcatg tcgtggcttt caacactcta
     1441 ttatttttga tgtcgttcac tgttttgtgt cttacaccag tatactcttt cttgcctggt
     1501 gtatactccg ttatctattt atacctgaca ttctacctga ctaacgatgt cagctttttg
     1561 gcacacattc aatggatggt aatgttcacc ccattggttc ccttctggat aacgatcgct
     1621 tacatcattt gcataagtac taaacatttc tattggtttt tctcaaatta tctaaagaga
     1681 agagttgtct ttaatggtgt cagcttttct accttcgaag aagctgcctt gtgtaccttt
     1741 ttgctcaaca aagaaatgta cttgaagttg agatctgacg ttttactccc gctaacccaa
     1801 tataacagat atttagcttt atacaacaag tacaaatact tctcgggtgc catggatact
     1861 acttcatacc gtgaagcggc ttgttgtcac cttgctaagg ctttgaatga tttctctaat
     1921 tccggttccg acgtcttgta tcaaccacct cagacgtcta ttacctccgc tgttttgcaa
     1981 ctcgtgccga gaggttctca ccatcatcac catcattaaa caggcccctt ttcctttgtc
     2041 gatatcatgt aattagttat gtcacgctta cattcacgcc ctcctcccac atccgctcta
     2101 accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt tttttaatag
     2161 ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacaaag
     2221 cggccgccga accactagtg cactgcagta cacggtttcc ttgaaatttt tttgattcgg
     2281 taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg gtatatatac
     2341 gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa ctgcacagaa
     2401 caaaaacgtg caggaaacga agataaatca tgtcgaaagc tacatataag gaacgtgctg
     2461 ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa aagcaaacaa
     2521 acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta gttgaagcat
     2581 taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat ttttccatgg
     2641 agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta ctcttcgagg
     2701 acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg ggtgtataca
     2761 gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca ggtattgtta
     2821 gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt ttgatgttag
     2881 cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt actgttgaca
     2941 ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac atgggtggaa
     3001 gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat gacaagggag
     3061 atgcattggg tcaacagtat agaaccgtgg atgatgtggt ttctacagga tctgacatta
     3121 ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt
     3181 acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac taaaaaactg
     3241 tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt taattatatc
     3301 agttattacc cgagtaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg
     3361 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     3421 gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca
     3481 acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat
     3541 gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg ttctacaaaa
     3601 atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt tttctccttt
     3661 gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt taaggttaga
     3721 agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc cacttcccgc
     3781 gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc atccccgatt
     3841 atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga
     3901 ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat atactacgta
     3961 taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt tcttactaca
     4021 atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag gtcgagttta
     4081 gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat agcacagaga
     4141 tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc aatattttag
     4201 tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcatca gagcgctttt
     4261 ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg aacttcccga
     4321 gcggccgcgt gttacaagct tgactccagc gtaactggac tgcaatcaac tcactggctc
     4381 accttcacgg gtgggccttt cttcggtaga aaatcaaagg atcttcttga gatccttttt
     4441 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4501 tgccggatca agagctacca actctttttc cgaggtaact ggcttcagca gagcgcagat
     4561 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     4621 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     4681 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     4741 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     4801 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     4861 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc caggggtaaa
     4921 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc atcgattttt
     4981 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcag aaaggcccac
     5041 ccgaaggtga gccaggtgat tacatttggg ccctcatcag aggttttcac cgtcatcacc
     5101 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     5161 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     5221 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactt accaatgctt
     5281 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5341 ccccgtcgtg tagataacta cgatgcggga gggcttacca tctggcccca gtgctgcaat
     5401 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5461 gagggccgag cgcagaagtg atcctgcaac tttatccgcc tccatccagt ctattaattg
     5521 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5581 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5641 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5701 cggtcctccg atcgttgcca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5761 agcactgcat aattctctta ctgtcatgcc atccgtgaga tgcttttctg tgactggtga
     5821 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5881 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5941 acgttcttcg gggcgtaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6001 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6061 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6121 aatactcatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
     6181 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat caagtcaaaa
     6241 gcctccggtc ggaggctttt gactttctgc tatggaggtc aggtatgatt taaatggtca
     6301 gttaga
//
