LOCUS       sequence-291491-        6423 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 leader protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6423)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6423)
  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-m4046362
FEATURES             Location/Qualifiers
     source          1..6423
                     /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             1018..1035
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2178..2196)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2547..3350
                     /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      3433..4415
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4493..4522
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4544..5131
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5143..5170)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5194..5385)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5386..6177,6178..6246))
                     /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     6009..6028
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6247..6349)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6350..6381)
                     /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 gaatctttgg tccctggatt taacgagaaa acccacgtac agctatccct tcccgtgttg
      541 caagttcgtg acgttctggt tcggggtttc ggtgattcag tcgaagaagt cttgagtgaa
      601 gccaggcaac atttaaagga tggtacttgt ggcttagttg aagtcgaaaa gggtgtgttg
      661 ccacaacttg agcaaccata cgttttcatt aagagatctg acgcaagaac agcgccacac
      721 ggtcatgtta tggttgaatt ggtcgctgaa ttggagggta tacaatatgg cagatcgggt
      781 gaaactttag gtgtattggt cccacacgta ggtgaaatcc ctgttgctta cagaaaagtt
      841 ttattaagaa agaatggtaa caagggtgct ggtggacact cctacggagc cgatctgaag
      901 tcttttgact tgggtgatga attgggtact gatccatatg aagacttcca agaaaattgg
      961 aacaccaaac atagctctgg tgttacgcgt gaattgatga gagaacttaa tggtggtctc
     1021 gtgccgagag gttctaaaat cgaggaaggt aagctagtca tttggataaa tggagataag
     1081 ggttacaacg gtttggcaga agtaggcaag aaatttgaaa aggacacagg tatcaaggtg
     1141 acggttgaac atccagacaa gttagaagaa aaattccctc aagttgctgc tactggtgat
     1201 ggtcccgaca ttattttctg ggcccacgat cgttttggtg gatatgctca gtcaggtttg
     1261 ctggctgaga tcaccccaga caaggccttc caagataaat tgtacccatt cacttgggat
     1321 gctgtcagat acaacggtaa gcttatagca tatccaattg ctgttgaagc cttatcgttg
     1381 atctacaata aggacttgtt acctaaccca ccaaagacat gggaagaaat tccagctttg
     1441 gataaagagc taaaggccaa gggaaaaagc gctttgatgt ttaacttaca agaaccttat
     1501 ttcacctggc cattaatcgc tgctgacggt ggttacgcat ttaaatacga aaacggtaag
     1561 tatgacatta aggatgtcgg cgttgataat gctggtgcaa aggctggttt gaccttcttg
     1621 gttgatttaa taaagaataa acacatgaac gctgacactg actactctat cgccgaagcc
     1681 gctttcaaca agggtgaaac tgctatgacc ataaatggtc catgggcctg gagtaacatt
     1741 gatacttcta aagtaaacta tggtgtcact gttttgccta cattcaaagg ccaaccatcc
     1801 aagccctttg tcggtgttct atccgctggc atcaatgctg cttctccaaa caaggagcta
     1861 gctaaagaat ttttggaaaa ttacttgtta actgacgaag gtttggaggc cgttaacaag
     1921 gacaagccat tgggtgctgt cgctttgaaa tcttacgaag aagaattggc taaggatccc
     1981 agaattgccg ctaccatgga gaacgcccaa aagggtgaaa ttatgccaaa catcccacaa
     2041 atgtcagcct tctggtacgc agttaggact gccgtgatta atgctgcttc cggtagacaa
     2101 accgtcgacg aagcattaaa agatgctcaa acgtaaacag gccccttttc ctttgtcgat
     2161 atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc cgctctaacc
     2221 gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt ttaatagtta
     2281 tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg tacaaagcgg
     2341 ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt gattcggtaa
     2401 tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta tatatacgca
     2461 tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg cacagaacaa
     2521 aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa cgtgctgcta
     2581 ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag caaacaaact
     2641 tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt gaagcattag
     2701 gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt tccatggagg
     2761 gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc ttcgaggaca
     2821 gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt gtatacagaa
     2881 tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt attgttagcg
     2941 gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg atgttagcag
     3001 aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact gttgacattg
     3061 cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg ggtggaagag
     3121 atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac aagggagatg
     3181 cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct gacattatta
     3241 ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt gaacgttaca
     3301 gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa aaaactgtat
     3361 tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa ttatatcagt
     3421 tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg caacgcgaga
     3481 gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg
     3541 aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg
     3601 cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca
     3661 acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc tacaaaaatg
     3721 catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt ctcctttgtg
     3781 cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa ggttagaaga
     3841 aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt
     3901 tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc cccgattata
     3961 ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg ttgatgattc
     4021 ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata ctacgtatag
     4081 gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct tactacaatt
     4141 tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc gagtttagat
     4201 gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc acagagatat
     4261 atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat attttagtag
     4321 ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag cgcttttggt
     4381 tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac ttcccgagcg
     4441 gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca ctggctcacc
     4501 ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat cctttttttc
     4561 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4621 cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag cgcagatacc
     4681 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4741 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4801 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     4861 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     4921 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4981 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag gggtaaacgc
     5041 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc gatttttgtg
     5101 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa ggcccacccg
     5161 aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt catcaccgaa
     5221 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc
     5281 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct
     5341 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc aatgcttaat
     5401 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5461 cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg ctgcaatgat
     5521 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccgggag
     5581 ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5641 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5701 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5761 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5821 tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5881 actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga ctggtgagta
     5941 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6001 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6061 ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6121 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6181 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6241 actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag
     6301 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa gtcaaaagcc
     6361 tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa atggtcagtt
     6421 aga
//
