LOCUS       sequence-291431-        7386 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 7386)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 7386)
  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-m4046160
FEATURES             Location/Qualifiers
     source          1..7386
                     /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"
     primer_bind     complement(3141..3159)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             3510..4313
                     /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      4396..5378
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      5456..5485
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      5507..6094
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(6106..6133)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6157..6348)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6349..7140,7141..7209))
                     /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     6972..6991
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7210..7312)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(7313..7344)
                     /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 gaggttctaa aatcgaggaa ggtaagctag tcatttggat aaatggagat
     2041 aagggttaca acggtttggc agaagtaggc aagaaatttg aaaaggacac aggtatcaag
     2101 gtgacggttg aacatccaga caagttagaa gaaaaattcc ctcaagttgc tgctactggt
     2161 gatggtcccg acattatttt ctgggcccac gatcgttttg gtggatatgc tcagtcaggt
     2221 ttgctggctg agatcacccc agacaaggcc ttccaagata aattgtaccc attcacttgg
     2281 gatgctgtca gatacaacgg taagcttata gcatatccaa ttgctgttga agccttatcg
     2341 ttgatctaca ataaggactt gttacctaac ccaccaaaga catgggaaga aattccagct
     2401 ttggataaag agctaaaggc caagggaaaa agcgctttga tgtttaactt acaagaacct
     2461 tatttcacct ggccattaat cgctgctgac ggtggttacg catttaaata cgaaaacggt
     2521 aagtatgaca ttaaggatgt cggcgttgat aatgctggtg caaaggctgg tttgaccttc
     2581 ttggttgatt taataaagaa taaacacatg aacgctgaca ctgactactc tatcgccgaa
     2641 gccgctttca acaagggtga aactgctatg accataaatg gtccatgggc ctggagtaac
     2701 attgatactt ctaaagtaaa ctatggtgtc actgttttgc ctacattcaa aggccaacca
     2761 tccaagccct ttgtcggtgt tctatccgct ggcatcaatg ctgcttctcc aaacaaggag
     2821 ctagctaaag aatttttgga aaattacttg ttaactgacg aaggtttgga ggccgttaac
     2881 aaggacaagc cattgggtgc tgtcgctttg aaatcttacg aagaagaatt ggctaaggat
     2941 cccagaattg ccgctaccat ggagaacgcc caaaagggtg aaattatgcc aaacatccca
     3001 caaatgtcag ccttctggta cgcagttagg actgccgtga ttaatgctgc ttccggtaga
     3061 caaaccgtcg acgaagcatt aaaagatgct caaacgtaaa caggcccctt ttcctttgtc
     3121 gatatcatgt aattagttat gtcacgctta cattcacgcc ctcctcccac atccgctcta
     3181 accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt tttttaatag
     3241 ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacaaag
     3301 cggccgccga accactagtg cactgcagta cacggtttcc ttgaaatttt tttgattcgg
     3361 taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg gtatatatac
     3421 gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa ctgcacagaa
     3481 caaaaacgtg caggaaacga agataaatca tgtcgaaagc tacatataag gaacgtgctg
     3541 ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa aagcaaacaa
     3601 acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta gttgaagcat
     3661 taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat ttttccatgg
     3721 agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta ctcttcgagg
     3781 acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg ggtgtataca
     3841 gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca ggtattgtta
     3901 gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt ttgatgttag
     3961 cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt actgttgaca
     4021 ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac atgggtggaa
     4081 gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat gacaagggag
     4141 atgcattggg tcaacagtat agaaccgtgg atgatgtggt ttctacagga tctgacatta
     4201 ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt
     4261 acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac taaaaaactg
     4321 tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt taattatatc
     4381 agttattacc cgagtaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg
     4441 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     4501 gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca
     4561 acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat
     4621 gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg ttctacaaaa
     4681 atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt tttctccttt
     4741 gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt taaggttaga
     4801 agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc cacttcccgc
     4861 gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc atccccgatt
     4921 atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga
     4981 ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat atactacgta
     5041 taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt tcttactaca
     5101 atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag gtcgagttta
     5161 gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat agcacagaga
     5221 tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc aatattttag
     5281 tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcatca gagcgctttt
     5341 ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg aacttcccga
     5401 gcggccgcgt gttacaagct tgactccagc gtaactggac tgcaatcaac tcactggctc
     5461 accttcacgg gtgggccttt cttcggtaga aaatcaaagg atcttcttga gatccttttt
     5521 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5581 tgccggatca agagctacca actctttttc cgaggtaact ggcttcagca gagcgcagat
     5641 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5701 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5761 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5821 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5881 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5941 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc caggggtaaa
     6001 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc atcgattttt
     6061 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcag aaaggcccac
     6121 ccgaaggtga gccaggtgat tacatttggg ccctcatcag aggttttcac cgtcatcacc
     6181 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     6241 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     6301 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactt accaatgctt
     6361 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6421 ccccgtcgtg tagataacta cgatgcggga gggcttacca tctggcccca gtgctgcaat
     6481 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6541 gagggccgag cgcagaagtg atcctgcaac tttatccgcc tccatccagt ctattaattg
     6601 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6661 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6721 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6781 cggtcctccg atcgttgcca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6841 agcactgcat aattctctta ctgtcatgcc atccgtgaga tgcttttctg tgactggtga
     6901 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6961 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7021 acgttcttcg gggcgtaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7081 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7141 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7201 aatactcatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
     7261 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat caagtcaaaa
     7321 gcctccggtc ggaggctttt gactttctgc tatggaggtc aggtatgatt taaatggtca
     7381 gttaga
//
