LOCUS       sequence-291626-        6223 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 6223)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6223)
  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-m4046708
FEATURES             Location/Qualifiers
     source          1..6223
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(1978..1996)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2347..3150
                     /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      3233..4215
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4293..4322
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4344..4931
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4943..4970)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4994..5185)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5186..5977,5978..6046))
                     /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     5809..5828
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6047..6149)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6150..6181)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgccc cacacaccat agcttcaaaa tgtttctact ccttttttac tcttccagat
       61 tttctcggac tccgcgcatc gccgtaccac ttcaaaacac ccaagcacag catactaaat
      121 ttcccctctt tcttcctcta gggtgtcgtt aattacccgt actaaaggtt tggaaaagaa
      181 aaaagagacc gcctcgtttc tttttcttcg tcgaaaaagg caataaaaat ttttatcacg
      241 tttctttttc ttgaaaattt ttttttttga tttttttctc tttcgatgac ctcccattga
      301 tatttaagtt aataaacggt cttcaatttc tcaagtttca gtttcatttt tcttgttcta
      361 ttacaacttt ttttacttct tgctcattag aaagaaagca tagcaatcta atctaagttt
      421 taattacaaa atgaagattg tcaacaattg gcttaaacag ttgataaagg ttactttagt
      481 atttttgttc gttgcagcca tcttctatct aattacacct gtccacgtga tgagtaagca
      541 taccgacttt tcctcagaga tcattggtta caaagctatc gatggcggag ttactagaga
      601 tattgcttcg acggacacct gtttcgctaa caagcacgcg gactttgata cttggttctc
      661 tcaaaggggt ggttcctaca caaacgacaa ggcctgcccc ttgattgcag ctgttatcac
      721 tagagaagtt ggtttcgtcg tgccaggttt accagggact atactgcgta ccaccaatgg
      781 tgattttttg catttcttac caagagtctt ctctgctgtt ggcaatattt gttatacccc
      841 tagcaaattg atcgaataca ctgattttgc cacctctgct tgtgtattgg ctgctgaatg
      901 cactattttc aaggatgcaa gtggtaagcc agttccatac tgttatgaca caaacgtcct
      961 cgaaggttct gttgcttacg aatccttgcg accggacact agatatgtgt taatggatgg
     1021 atcaatcatt caatttccaa acacatacct tgaaggttct gttcgggttg tcaccacctt
     1081 cgactccgag tactgtcgcc acggtacttg tgaaagatca gaagccggtg tctgtgtttc
     1141 taccagcggt agatgggtgt tgaataacga ttattaccgt tctcttccag gtgtcttctg
     1201 cggtgtcgat gctgttaatc tattgactaa catgtttacc cctttgatcc aaccaatagg
     1261 tgcgctggac atttccgcat ctattgtcgc cggtggaatc gttgctatcg ttgttacttg
     1321 tttggcttac tacttcatga gatttagaag agcctttggg gagtattcgc atgtcgtggc
     1381 tttcaacact ctattatttt tgatgtcgtt cactgttttg tgtcttacac cagtatactc
     1441 tttcttgcct ggtgtatact ccgttatcta tttatacctg acattctacc tgactaacga
     1501 tgtcagcttt ttggcacaca ttcaatggat ggtaatgttc accccattgg ttcccttctg
     1561 gataacgatc gcttacatca tttgcataag tactaaacat ttctattggt ttttctcaaa
     1621 ttatctaaag agaagagttg tctttaatgg tgtcagcttt tctaccttcg aagaagctgc
     1681 cttgtgtacc tttttgctca acaaagaaat gtacttgaag ttgagatctg acgttttact
     1741 cccgctaacc caatataaca gatatttagc tttatacaac aagtacaaat acttctcggg
     1801 tgccatggat actacttcat accgtgaagc ggcttgttgt caccttgcta aggctttgaa
     1861 tgatttctct aattccggtt ccgacgtctt gtatcaacca cctcagacgt ctattacctc
     1921 cgctgttttg caataaacag gccccttttc ctttgtcgat atcatgtaat tagttatgtc
     1981 acgcttacat tcacgccctc ctcccacatc cgctctaacc gaaaaggaag gagttagaca
     2041 acctgaagtc taggtcccta tttatttttt ttaatagtta tgttagtatt aagaacgtta
     2101 tttatatttc aaatttttct tttttttctg tacaaagcgg ccgccgaacc actagtgcac
     2161 tgcagtacac ggtttccttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa
     2221 cgaaggaagg agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca
     2281 tgaaattgcc cagtattctt aacccaactg cacagaacaa aaacgtgcag gaaacgaaga
     2341 taaatcatgt cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct
     2401 gccaagctat ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt
     2461 cgtaccacca aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta
     2521 aaaacacatg tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag
     2581 gcattatccg ccaagtacaa ttttttactc ttcgaggaca gaaaatttgc tgacattggt
     2641 aatacagtca aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt
     2701 acgaatgcac acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa
     2761 gaagtaacaa aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc
     2821 ctatctactg gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt
     2881 gttatcggct ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg
     2941 attatgacac ccggtgtggg tttagatgac aagggagatg cattgggtca acagtataga
     3001 accgtggatg atgtggtttc tacaggatct gacattatta ttgttggaag aggactattt
     3061 gcaaagggaa gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca
     3121 tatttgagaa gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata
     3181 ctaaactcac aaattagagc ttcaatttaa ttatatcagt tattacccga gtaacgaagc
     3241 atctgtgctt cattttgtag aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa
     3301 gaatctgagc tgcattttta cagaacagaa atgcaacgcg aaagcgctat tttaccaacg
     3361 aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa
     3421 acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgagagc gctattttac
     3481 caacaaagaa tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt
     3541 tctaacaaag catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct
     3601 tgataacttt ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt
     3661 ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct
     3721 gcgggtgcat tttttcaaga taaaggcatc cccgattata ttctataccg atgtggattg
     3781 cgcatacttt gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat
     3841 gaacggtttc ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta
     3901 ttgttttcga ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata
     3961 ctagagataa acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag
     4021 gtggatgggt aggttatata gggatatagc acagagatat atagcaaaga gatacttttg
     4081 agcaatgttt gtggaagcgg tattcgcaat attttagtag ctcgttacag tccggtgcgt
     4141 ttttggtttt ttgaaagtgc gtcatcagag cgcttttggt tttcaaaagc gctctgaagt
     4201 tcctatactt tctagctaga gaataggaac ttcccgagcg gccgcgtgtt acaagcttga
     4261 ctccagcgta actggactgc aatcaactca ctggctcacc ttcacgggtg ggcctttctt
     4321 cggtagaaaa tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4381 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4441 ctttttccga ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4501 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4561 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4621 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4681 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4741 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4801 gaacaggaga gcgcacgagg gagcttccag gggtaaacgc ctggtatctt tatagtcctg
     4861 tcgggtttcg ccacctctga cttgagcatc gatttttgtg atgctcgtca ggggggcgga
     4921 gcctatggaa aaacgccagc aacgcagaaa ggcccacccg aaggtgagcc aggtgattac
     4981 atttgggccc tcatcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     5041 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
     5101 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     5161 gggcggtttt ttcctgtttg gtcacttacc aatgcttaat cagtgaggca cctatctcag
     5221 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5281 tgcgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5341 cggctccaga tttatcagca ataaaccagc cagccgggag ggccgagcgc agaagtgatc
     5401 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5461 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5521 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5581 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgccagaa
     5641 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5701 tcatgccatc cgtgagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5761 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5821 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgtaaactct
     5881 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5941 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6001 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatttta gcttccttag
     6061 ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt
     6121 gaaagttgga acctcttacg tgccgatcaa gtcaaaagcc tccggtcgga ggcttttgac
     6181 tttctgctat ggaggtcagg tatgatttaa atggtcagtt aga
//
