LOCUS       sequence-298845-        5343 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 5343)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5343)
  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-m4046187
FEATURES             Location/Qualifiers
     source          1..5343
                     /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"
     CDS             1036..1053
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1098..1116)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1467..2270
                     /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      2353..3335
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3413..3442
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3464..4051
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4063..4090)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4114..4305)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4306..5097,5098..5166))
                     /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     4929..4948
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5167..5269)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5270..5301)
                     /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 gttctcacca tcatcaccat cattaaacag gccccttttc ctttgtcgat
     1081 atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc cgctctaacc
     1141 gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt ttaatagtta
     1201 tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg tacaaagcgg
     1261 ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt gattcggtaa
     1321 tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta tatatacgca
     1381 tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg cacagaacaa
     1441 aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa cgtgctgcta
     1501 ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag caaacaaact
     1561 tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt gaagcattag
     1621 gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt tccatggagg
     1681 gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc ttcgaggaca
     1741 gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt gtatacagaa
     1801 tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt attgttagcg
     1861 gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg atgttagcag
     1921 aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact gttgacattg
     1981 cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg ggtggaagag
     2041 atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac aagggagatg
     2101 cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct gacattatta
     2161 ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt gaacgttaca
     2221 gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa aaaactgtat
     2281 tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa ttatatcagt
     2341 tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg caacgcgaga
     2401 gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg
     2461 aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg
     2521 cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca
     2581 acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc tacaaaaatg
     2641 catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt ctcctttgtg
     2701 cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa ggttagaaga
     2761 aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt
     2821 tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc cccgattata
     2881 ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg ttgatgattc
     2941 ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata ctacgtatag
     3001 gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct tactacaatt
     3061 tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc gagtttagat
     3121 gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc acagagatat
     3181 atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat attttagtag
     3241 ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag cgcttttggt
     3301 tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac ttcccgagcg
     3361 gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca ctggctcacc
     3421 ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat cctttttttc
     3481 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3541 cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag cgcagatacc
     3601 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3661 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3721 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3781 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3841 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3901 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag gggtaaacgc
     3961 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc gatttttgtg
     4021 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa ggcccacccg
     4081 aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt catcaccgaa
     4141 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc
     4201 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct
     4261 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc aatgcttaat
     4321 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4381 cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg ctgcaatgat
     4441 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccgggag
     4501 ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4561 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4621 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4681 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4741 tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4801 actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga ctggtgagta
     4861 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4921 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4981 ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5041 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5101 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5161 actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag
     5221 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa gtcaaaagcc
     5281 tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa atggtcagtt
     5341 aga
//
