LOCUS       sequence-298296-        6201 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 nsp4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6201)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6201)
  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..6201
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(1956..1974)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2325..3128
                     /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      3211..4193
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4271..4300
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4322..4909
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4921..4948)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4972..5163)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5164..5955,5956..6024))
                     /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     5787..5806
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6025..6127)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6128..6159)
                     /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 tcgaaaagtt tctttttctt gaaaattttt
      241 ttttttgatt tttttctctt tcgatgacct cccattgata tttaagttaa taaacggtct
      301 tcaatttctc aagtttcagt ttcatttttc ttgttctatt acaacttttt ttacttcttg
      361 ctcattagaa agaaagcata gcaatctaat ctaagtttta attacaaaat gaagattgtc
      421 aacaattggc ttaaacagtt gataaaggtt actttagtat ttttgttcgt tgcagccatc
      481 ttctatctaa ttacacctgt ccacgtgatg agtaagcata ccgacttttc ctcagagatc
      541 attggttaca aagctatcga tggcggagtt actagagata ttgcttcgac ggacacctgt
      601 ttcgctaaca agcacgcgga ctttgatact tggttctctc aaaggggtgg ttcctacaca
      661 aacgacaagg cctgcccctt gattgcagct gttatcacta gagaagttgg tttcgtcgtg
      721 ccaggtttac cagggactat actgcgtacc accaatggtg attttttgca tttcttacca
      781 agagtcttct ctgctgttgg caatatttgt tataccccta gcaaattgat cgaatacact
      841 gattttgcca cctctgcttg tgtattggct gctgaatgca ctattttcaa ggatgcaagt
      901 ggtaagccag ttccatactg ttatgacaca aacgtcctcg aaggttctgt tgcttacgaa
      961 tccttgcgac cggacactag atatgtgtta atggatggat caatcattca atttccaaac
     1021 acataccttg aaggttctgt tcgggttgtc accaccttcg actccgagta ctgtcgccac
     1081 ggtacttgtg aaagatcaga agccggtgtc tgtgtttcta ccagcggtag atgggtgttg
     1141 aataacgatt attaccgttc tcttccaggt gtcttctgcg gtgtcgatgc tgttaatcta
     1201 ttgactaaca tgtttacccc tttgatccaa ccaataggtg cgctggacat ttccgcatct
     1261 attgtcgccg gtggaatcgt tgctatcgtt gttacttgtt tggcttacta cttcatgaga
     1321 tttagaagag cctttgggga gtattcgcat gtcgtggctt tcaacactct attatttttg
     1381 atgtcgttca ctgttttgtg tcttacacca gtatactctt tcttgcctgg tgtatactcc
     1441 gttatctatt tatacctgac attctacctg actaacgatg tcagcttttt ggcacacatt
     1501 caatggatgg taatgttcac cccattggtt cccttctgga taacgatcgc ttacatcatt
     1561 tgcataagta ctaaacattt ctattggttt ttctcaaatt atctaaagag aagagttgtc
     1621 tttaatggtg tcagcttttc taccttcgaa gaagctgcct tgtgtacctt tttgctcaac
     1681 aaagaaatgt acttgaagtt gagatctgac gttttactcc cgctaaccca atataacaga
     1741 tatttagctt tatacaacaa gtacaaatac ttctcgggtg ccatggatac tacttcatac
     1801 cgtgaagcgg cttgttgtca ccttgctaag gctttgaatg atttctctaa ttccggttcc
     1861 gacgtcttgt atcaaccacc tcagacgtct attacctccg ctgttttgca ataaacaggc
     1921 cccttttcct ttgtcgatat catgtaatta gttatgtcac gcttacattc acgccctcct
     1981 cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt
     2041 tatttttttt aatagttatg ttagtattaa gaacgttatt tatatttcaa atttttcttt
     2101 tttttctgta caaagcggcc gccgaaccac tagtgcactg cagtacacgg tttccttgaa
     2161 atttttttga ttcggtaatc tccgaacaga aggaagaacg aaggaaggag cacagactta
     2221 gattggtata tatacgcata tgtagtgttg aagaaacatg aaattgccca gtattcttaa
     2281 cccaactgca cagaacaaaa acgtgcagga aacgaagata aatcatgtcg aaagctacat
     2341 ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt aatatcatgc
     2401 acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag gaattactgg
     2461 agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg gatatcttga
     2521 ctgatttttc catggagggc acagttaagc cgctaaaggc attatccgcc aagtacaatt
     2581 ttttactctt cgaggacaga aaatttgctg acattggtaa tacagtcaaa ttgcagtact
     2641 ctgcgggtgt atacagaata gcagaatggg cagacattac gaatgcacac ggtgtggtgg
     2701 gcccaggtat tgttagcggt ttgaagcagg cggcagaaga agtaacaaag gaacctagag
     2761 gccttttgat gttagcagaa ttgtcatgca agggctccct atctactgga gaatatacta
     2821 agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt attgctcaaa
     2881 gagacatggg tggaagagat gaaggttacg attggttgat tatgacaccc ggtgtgggtt
     2941 tagatgacaa gggagatgca ttgggtcaac agtatagaac cgtggatgat gtggtttcta
     3001 caggatctga cattattatt gttggaagag gactatttgc aaagggaagg gatgctaagg
     3061 tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga tgcggccagc
     3121 aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa attagagctt
     3181 caatttaatt atatcagtta ttacccgagt aacgaagcat ctgtgcttca ttttgtagaa
     3241 caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca
     3301 gaacagaaat gcaacgcgaa agcgctattt taccaacgaa gaatctgtgc ttcatttttg
     3361 taaaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt
     3421 ttacagaaca gaaatgcaac gcgagagcgc tattttacca acaaagaatc tatacttctt
     3481 ttttgttcta caaaaatgca tcccgagagc gctatttttc taacaaagca tcttagatta
     3541 ctttttttct cctttgtgcg ctctataatg cagtctcttg ataacttttt gcactgtagg
     3601 tccgttaagg ttagaagaag gctactttgg tgtctatttt ctcttccata aaaaaagcct
     3661 gactccactt cccgcgttta ctgattacta gcgaagctgc gggtgcattt tttcaagata
     3721 aaggcatccc cgattatatt ctataccgat gtggattgcg catactttgt gaacagaaag
     3781 tgatagcgtt gatgattctt cattggtcag aaaattatga acggtttctt ctattttgtc
     3841 tctatatact acgtatagga aatgtttaca ttttcgtatt gttttcgatt cactctatga
     3901 atagttctta ctacaatttt tttgtctaaa gagtaatact agagataaac ataaaaaatg
     3961 tagaggtcga gtttagatgc aagttcaagg agcgaaaggt ggatgggtag gttatatagg
     4021 gatatagcac agagatatat agcaaagaga tacttttgag caatgtttgt ggaagcggta
     4081 ttcgcaatat tttagtagct cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt
     4141 catcagagcg cttttggttt tcaaaagcgc tctgaagttc ctatactttc tagctagaga
     4201 ataggaactt cccgagcggc cgcgtgttac aagcttgact ccagcgtaac tggactgcaa
     4261 tcaactcact ggctcacctt cacgggtggg cctttcttcg gtagaaaatc aaaggatctt
     4321 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4381 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgagg taactggctt
     4441 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     4501 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4561 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4621 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4681 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     4741 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     4801 gcttccaggg gtaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     4861 tgagcatcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     4921 cgcagaaagg cccacccgaa ggtgagccag gtgattacat ttgggccctc atcagaggtt
     4981 ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg
     5041 aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag
     5101 cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt
     5161 cacttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5221 catagttgcc tgactccccg tcgtgtagat aactacgatg cgggagggct taccatctgg
     5281 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5341 aaaccagcca gccgggaggg ccgagcgcag aagtgatcct gcaactttat ccgcctccat
     5401 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5461 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5521 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5581 agcggttagc tccttcggtc ctccgatcgt tgccagaagt aagttggccg cagtgttatc
     5641 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg tgagatgctt
     5701 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5761 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5821 gctcatcatt ggaaaacgtt cttcggggcg taaactctca aggatcttac cgctgttgag
     5881 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5941 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6001 gacacggaaa tgttgaatac tcattttagc ttccttagct cctgaaaatc tcgataactc
     6061 aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg
     6121 ccgatcaagt caaaagcctc cggtcggagg cttttgactt tctgctatgg aggtcaggta
     6181 tgatttaaat ggtcagttag a
//
