LOCUS       sequence-297558-        6270 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 6270)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6270)
  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-m4046697
FEATURES             Location/Qualifiers
     source          1..6270
                     /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"
     primer_bind     complement(2025..2043)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2394..3197
                     /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      3280..4262
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4340..4369
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4391..4978
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4990..5017)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5041..5232)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5233..6024,6025..6093))
                     /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     5856..5875
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6094..6196)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6197..6228)
                     /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 taaacaggcc ccttttcctt tgtcgatatc atgtaattag ttatgtcacg cttacattca
     2041 cgccctcctc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag
     2101 gtccctattt atttttttta atagttatgt tagtattaag aacgttattt atatttcaaa
     2161 tttttctttt ttttctgtac aaagcggccg ccgaaccact agtgcactgc agtacacggt
     2221 ttccttgaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga aggaaggagc
     2281 acagacttag attggtatat atacgcatat gtagtgttga agaaacatga aattgcccag
     2341 tattcttaac ccaactgcac agaacaaaaa cgtgcaggaa acgaagataa atcatgtcga
     2401 aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc aagctattta
     2461 atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt accaccaagg
     2521 aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa acacatgtgg
     2581 atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca ttatccgcca
     2641 agtacaattt tttactcttc gaggacagaa aatttgctga cattggtaat acagtcaaat
     2701 tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg aatgcacacg
     2761 gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa gtaacaaagg
     2821 aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta tctactggag
     2881 aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt atcggcttta
     2941 ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt atgacacccg
     3001 gtgtgggttt agatgacaag ggagatgcat tgggtcaaca gtatagaacc gtggatgatg
     3061 tggtttctac aggatctgac attattattg ttggaagagg actatttgca aagggaaggg
     3121 atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat ttgagaagat
     3181 gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta aactcacaaa
     3241 ttagagcttc aatttaatta tatcagttat tacccgagta acgaagcatc tgtgcttcat
     3301 tttgtagaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc
     3361 atttttacag aacagaaatg caacgcgaaa gcgctatttt accaacgaag aatctgtgct
     3421 tcatttttgt aaaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga
     3481 gctgcatttt tacagaacag aaatgcaacg cgagagcgct attttaccaa caaagaatct
     3541 atacttcttt tttgttctac aaaaatgcat cccgagagcg ctatttttct aacaaagcat
     3601 cttagattac tttttttctc ctttgtgcgc tctataatgc agtctcttga taactttttg
     3661 cactgtaggt ccgttaaggt tagaagaagg ctactttggt gtctattttc tcttccataa
     3721 aaaaagcctg actccacttc ccgcgtttac tgattactag cgaagctgcg ggtgcatttt
     3781 ttcaagataa aggcatcccc gattatattc tataccgatg tggattgcgc atactttgtg
     3841 aacagaaagt gatagcgttg atgattcttc attggtcaga aaattatgaa cggtttcttc
     3901 tattttgtct ctatatacta cgtataggaa atgtttacat tttcgtattg ttttcgattc
     3961 actctatgaa tagttcttac tacaattttt ttgtctaaag agtaatacta gagataaaca
     4021 taaaaaatgt agaggtcgag tttagatgca agttcaagga gcgaaaggtg gatgggtagg
     4081 ttatataggg atatagcaca gagatatata gcaaagagat acttttgagc aatgtttgtg
     4141 gaagcggtat tcgcaatatt ttagtagctc gttacagtcc ggtgcgtttt tggttttttg
     4201 aaagtgcgtc atcagagcgc ttttggtttt caaaagcgct ctgaagttcc tatactttct
     4261 agctagagaa taggaacttc ccgagcggcc gcgtgttaca agcttgactc cagcgtaact
     4321 ggactgcaat caactcactg gctcaccttc acgggtgggc ctttcttcgg tagaaaatca
     4381 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4441 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaggt
     4501 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4561 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4621 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4681 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4741 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4801 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4861 cacgagggag cttccagggg taaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4921 cctctgactt gagcatcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4981 cgccagcaac gcagaaaggc ccacccgaag gtgagccagg tgattacatt tgggccctca
     5041 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc
     5101 gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt
     5161 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc
     5221 ctgtttggtc acttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5281 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatgc gggagggctt
     5341 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5401 atcagcaata aaccagccag ccgggagggc cgagcgcaga agtgatcctg caactttatc
     5461 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5521 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5581 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5641 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gccagaagta agttggccgc
     5701 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5761 gagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5821 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5881 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcgt aaactctcaa ggatcttacc
     5941 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6001 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6061 aataagggcg acacggaaat gttgaatact cattttagct tccttagctc ctgaaaatct
     6121 cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc
     6181 tcttacgtgc cgatcaagtc aaaagcctcc ggtcggaggc ttttgacttt ctgctatgga
     6241 ggtcaggtat gatttaaatg gtcagttaga
//
