LOCUS       sequence-291608-        6217 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 6217)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6217)
  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-m4046667
FEATURES             Location/Qualifiers
     source          1..6217
                     /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(1972..1990)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2341..3144
                     /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      3227..4209
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4287..4316
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4338..4925
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4937..4964)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4988..5179)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5180..5971,5972..6040))
                     /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     5803..5822
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6041..6143)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6144..6175)
                     /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 atgattgtta ataattggtt gaagcagtta attaaagtta cacttgtgtt
      481 cctttttgtt gctgctattt tctatttaat aacacctgtt catgtcatgt ctaaacatac
      541 tgacttttca agtgaaatca taggatacaa ggctattgat ggtggtgtca ctcgtgacat
      601 agcatctaca gatacttgtt ttgctaacaa acatgctgat tttgacacat ggtttagcca
      661 gcgtggtggt agttatacta atgacaaagc ttgcccattg attgctgcag tcataacaag
      721 agaagtgggt tttgtcgtgc ctggtttgcc tggcacgata ttacgcacaa ctaatggtga
      781 ctttttgcat ttcttaccta gagtttttag tgcagttggt aacatctgtt acacaccatc
      841 aaaacttata gagtacactg actttgcaac atcagcttgt gttttggctg ctgaatgtac
      901 aatttttaaa gatgcttctg gtaagccagt accatattgt tatgatacca atgtactaga
      961 aggttctgtt gcttatgaaa gtttacgccc tgacacacgt tatgtgctca tggatggctc
     1021 tattattcaa tttcctaaca cctaccttga aggttctgtt agagtggtaa caacttttga
     1081 ttctgagtac tgtaggcacg gcacttgtga aagatcagaa gctggtgttt gtgtatctac
     1141 tagtggtaga tgggtactta acaatgatta ttacagatct ttaccaggag ttttctgtgg
     1201 tgtagatgct gtaaatttac ttactaatat gtttacacca ctaattcaac ctattggtgc
     1261 tttggacata tcagcatcta tagtagctgg tggtattgta gctatcgtag taacatgcct
     1321 tgcctactat tttatgaggt ttagaagagc ttttggtgaa tacagtcatg tagttgcctt
     1381 taatacttta ctattcctta tgtcattcac tgtactctgt ttaacaccag tttactcatt
     1441 cttacctggt gtttattctg ttatttactt gtacttgaca ttttatctta ctaatgatgt
     1501 ttctttttta gcacatattc agtggatggt tatgttcaca cctttagtac ctttctggat
     1561 aacaattgct tatatcattt gtatttccac aaagcatttc tattggttct ttagtaatta
     1621 cctaaagaga cgtgtagtct ttaatggtgt ttcctttagt acttttgaag aagctgcgct
     1681 gtgcaccttt ttgttaaata aagaaatgta tctaaagttg cgtagtgatg tgctattacc
     1741 tcttacgcaa tataatagat acttagctct ttataataag tacaagtatt ttagtggagc
     1801 aatggataca actagctaca gagaagctgc ttgttgtcat ctcgcaaagg ctctcaatga
     1861 cttcagtaac tcaggttctg atgttcttta ccaaccacca caaacctcta tcacctcagc
     1921 tgttttgtaa acaggcccct tttcctttgt cgatatcatg taattagtta tgtcacgctt
     1981 acattcacgc cctcctccca catccgctct aaccgaaaag gaaggagtta gacaacctga
     2041 agtctaggtc cctatttatt ttttttaata gttatgttag tattaagaac gttatttata
     2101 tttcaaattt ttcttttttt tctgtacaaa gcggccgccg aaccactagt gcactgcagt
     2161 acacggtttc cttgaaattt ttttgattcg gtaatctccg aacagaagga agaacgaagg
     2221 aaggagcaca gacttagatt ggtatatata cgcatatgta gtgttgaaga aacatgaaat
     2281 tgcccagtat tcttaaccca actgcacaga acaaaaacgt gcaggaaacg aagataaatc
     2341 atgtcgaaag ctacatataa ggaacgtgct gctactcatc ctagtcctgt tgctgccaag
     2401 ctatttaata tcatgcacga aaagcaaaca aacttgtgtg cttcattgga tgttcgtacc
     2461 accaaggaat tactggagtt agttgaagca ttaggtccca aaatttgttt actaaaaaca
     2521 catgtggata tcttgactga tttttccatg gagggcacag ttaagccgct aaaggcatta
     2581 tccgccaagt acaatttttt actcttcgag gacagaaaat ttgctgacat tggtaataca
     2641 gtcaaattgc agtactctgc gggtgtatac agaatagcag aatgggcaga cattacgaat
     2701 gcacacggtg tggtgggccc aggtattgtt agcggtttga agcaggcggc agaagaagta
     2761 acaaaggaac ctagaggcct tttgatgtta gcagaattgt catgcaaggg ctccctatct
     2821 actggagaat atactaaggg tactgttgac attgcgaaga gcgacaaaga ttttgttatc
     2881 ggctttattg ctcaaagaga catgggtgga agagatgaag gttacgattg gttgattatg
     2941 acacccggtg tgggtttaga tgacaaggga gatgcattgg gtcaacagta tagaaccgtg
     3001 gatgatgtgg tttctacagg atctgacatt attattgttg gaagaggact atttgcaaag
     3061 ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag caggctggga agcatatttg
     3121 agaagatgcg gccagcaaaa ctaaaaaact gtattataag taaatgcatg tatactaaac
     3181 tcacaaatta gagcttcaat ttaattatat cagttattac ccgagtaacg aagcatctgt
     3241 gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct
     3301 gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg ctattttacc aacgaagaat
     3361 ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag
     3421 aatctgagct gcatttttac agaacagaaa tgcaacgcga gagcgctatt ttaccaacaa
     3481 agaatctata cttctttttt gttctacaaa aatgcatccc gagagcgcta tttttctaac
     3541 aaagcatctt agattacttt ttttctcctt tgtgcgctct ataatgcagt ctcttgataa
     3601 ctttttgcac tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc tattttctct
     3661 tccataaaaa aagcctgact ccacttcccg cgtttactga ttactagcga agctgcgggt
     3721 gcattttttc aagataaagg catccccgat tatattctat accgatgtgg attgcgcata
     3781 ctttgtgaac agaaagtgat agcgttgatg attcttcatt ggtcagaaaa ttatgaacgg
     3841 tttcttctat tttgtctcta tatactacgt ataggaaatg tttacatttt cgtattgttt
     3901 tcgattcact ctatgaatag ttcttactac aatttttttg tctaaagagt aatactagag
     3961 ataaacataa aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg aaaggtggat
     4021 gggtaggtta tatagggata tagcacagag atatatagca aagagatact tttgagcaat
     4081 gtttgtggaa gcggtattcg caatatttta gtagctcgtt acagtccggt gcgtttttgg
     4141 ttttttgaaa gtgcgtcatc agagcgcttt tggttttcaa aagcgctctg aagttcctat
     4201 actttctagc tagagaatag gaacttcccg agcggccgcg tgttacaagc ttgactccag
     4261 cgtaactgga ctgcaatcaa ctcactggct caccttcacg ggtgggcctt tcttcggtag
     4321 aaaatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4381 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4441 ccgaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4501 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4561 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4621 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4681 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4741 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4801 gagagcgcac gagggagctt ccaggggtaa acgcctggta tctttatagt cctgtcgggt
     4861 ttcgccacct ctgacttgag catcgatttt tgtgatgctc gtcagggggg cggagcctat
     4921 ggaaaaacgc cagcaacgca gaaaggccca cccgaaggtg agccaggtga ttacatttgg
     4981 gccctcatca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     5041 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     5101 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     5161 ttttttcctg tttggtcact taccaatgct taatcagtga ggcacctatc tcagcgatct
     5221 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatgcggg
     5281 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     5341 cagatttatc agcaataaac cagccagccg ggagggccga gcgcagaagt gatcctgcaa
     5401 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     5461 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     5521 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5581 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgcc agaagtaagt
     5641 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5701 catccgtgag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5761 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5821 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgtaaa ctctcaagga
     5881 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5941 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     6001 aaaagggaat aagggcgaca cggaaatgtt gaatactcat tttagcttcc ttagctcctg
     6061 aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt
     6121 tggaacctct tacgtgccga tcaagtcaaa agcctccggt cggaggcttt tgactttctg
     6181 ctatggaggt caggtatgat ttaaatggtc agttaga
//
