LOCUS       sequence-297711-        6364 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 leader protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6364)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6364)
  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-m4046392
FEATURES             Location/Qualifiers
     source          1..6364
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        4..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             968..985
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2128..2146)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2488..3291
                     /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      3374..4356
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4434..4463
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4485..5072
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5084..5111)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5135..5326)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5327..6118,6119..6187))
                     /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     5950..5969
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6188..6290)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6291..6322)
                     /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 atggagagcc ttgtccctgg tttcaacgag aaaacacacg tccaactcag
      481 tttgcctgtt ttacaggttc gcgacgtgct cgtacgtggc tttggagact ccgtggagga
      541 ggtcttatca gaggcacgtc aacatcttaa agatggcact tgtggcttag tagaagttga
      601 aaaaggcgtt ttgcctcaac ttgaacagcc ctatgtgttc atcaaacgtt cggatgctcg
      661 aactgcacct catggtcatg ttatggttga gctggtagca gaactcgaag gcattcagta
      721 cggtcgtagt ggtgagacac ttggtgtcct tgtccctcat gtgggcgaaa taccagtggc
      781 ttaccgcaag gttcttcttc gtaagaacgg taataaagga gctggtggcc atagttacgg
      841 cgccgatcta aagtcatttg acttaggcga cgagcttggc actgatcctt atgaagattt
      901 tcaagaaaac tggaacacta aacatagcag tggtgttacc cgtgaactca tgcgtgagct
      961 taacggactc gtgccgagag gttctaaaat cgaggaaggt aagctagtca tttggataaa
     1021 tggagataag ggttacaacg gtttggcaga agtaggcaag aaatttgaaa aggacacagg
     1081 tatcaaggtg acggttgaac atccagacaa gttagaagaa aaattccctc aagttgctgc
     1141 tactggtgat ggtcccgaca ttattttctg ggcccacgat cgttttggtg gatatgctca
     1201 gtcaggtttg ctggctgaga tcaccccaga caaggccttc caagataaat tgtacccatt
     1261 cacttgggat gctgtcagat acaacggtaa gcttatagca tatccaattg ctgttgaagc
     1321 cttatcgttg atctacaata aggacttgtt acctaaccca ccaaagacat gggaagaaat
     1381 tccagctttg gataaagagc taaaggccaa gggaaaaagc gctttgatgt ttaacttaca
     1441 agaaccttat ttcacctggc cattaatcgc tgctgacggt ggttacgcat ttaaatacga
     1501 aaacggtaag tatgacatta aggatgtcgg cgttgataat gctggtgcaa aggctggttt
     1561 gaccttcttg gttgatttaa taaagaataa acacatgaac gctgacactg actactctat
     1621 cgccgaagcc gctttcaaca agggtgaaac tgctatgacc ataaatggtc catgggcctg
     1681 gagtaacatt gatacttcta aagtaaacta tggtgtcact gttttgccta cattcaaagg
     1741 ccaaccatcc aagccctttg tcggtgttct atccgctggc atcaatgctg cttctccaaa
     1801 caaggagcta gctaaagaat ttttggaaaa ttacttgtta actgacgaag gtttggaggc
     1861 cgttaacaag gacaagccat tgggtgctgt cgctttgaaa tcttacgaag aagaattggc
     1921 taaggatccc agaattgccg ctaccatgga gaacgcccaa aagggtgaaa ttatgccaaa
     1981 catcccacaa atgtcagcct tctggtacgc agttaggact gccgtgatta atgctgcttc
     2041 cggtagacaa accgtcgacg aagcattaaa agatgctcaa acgtaaacag gccccttttc
     2101 ctttgtcgat atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc
     2161 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
     2221 ttaatagtta tgttagtatt aagaacgttt caaatttttc ttttttttct gtacaaagcg
     2281 gccgccgaac cactagtgca ctgcagtaca cggtttcctt gaaatttttt tgattcggta
     2341 atctccgaac agaaggaaga acgaaggaag gagcacagac ttagattggt atatatacgc
     2401 atatgtagtg ttgaagaaac atgaaattgc ccagtattct taacccaact gcacagaaca
     2461 aaaacgtgca ggaaacgaag ataaatcatg tcgaaagcta catataagga acgtgctgct
     2521 actcatccta gtcctgttgc tgccaagcta tttaatatca tgcacgaaaa gcaaacaaac
     2581 ttgtgtgctt cattggatgt tcgtaccacc aaggaattac tggagttagt tgaagcatta
     2641 ggtcccaaaa tttgtttact aaaaacacat gtggatatct tgactgattt ttccatggag
     2701 ggcacagtta agccgctaaa ggcattatcc gccaagtaca attttttact cttcgaggac
     2761 agaaaatttg ctgacattgg taatacagtc aaattgcagt actctgcggg tgtatacaga
     2821 atagcagaat gggcagacat tacgaatgca cacggtgtgg tgggcccagg tattgttagc
     2881 ggtttgaagc aggcggcaga agaagtaaca aaggaaccta gaggcctttt gatgttagca
     2941 gaattgtcat gcaagggctc cctatctact ggagaatata ctaagggtac tgttgacatt
     3001 gcgaagagcg acaaagattt tgttatcggc tttattgctc aaagagacat gggtggaaga
     3061 gatgaaggtt acgattggtt gattatgaca cccggtgtgg gtttagatga caagggagat
     3121 gcattgggtc aacagtatag aaccgtggat gatgtggttt ctacaggatc tgacattatt
     3181 attgttggaa gaggactatt tgcaaaggga agggatgcta aggtagaggg tgaacgttac
     3241 agaaaagcag gctgggaagc atatttgaga agatgcggcc agcaaaacta aaaaactgta
     3301 ttataagtaa atgcatgtat actaaactca caaattagag cttcaattta attatatcag
     3361 ttattacccg agtaacgaag catctgtgct tcattttgta gaacaaaaat gcaacgcgag
     3421 agcgctaatt tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc
     3481 gaaagcgcta ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac
     3541 gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc
     3601 aacgcgagag cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat
     3661 gcatcccgag agcgctatty ttctaacaaa gcatcttaga ttactttttt tctcctttgt
     3721 gcgctctata atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag
     3781 aaggctactt tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt
     3841 ttactgatta ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat
     3901 attctatacc gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt
     3961 cttcattggt cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata
     4021 ggaaatgttt acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat
     4081 ttttttgtct aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga
     4141 tgcaagttca aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata
     4201 tatagcaaag agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tattttagta
     4261 gctcgttaca gtccggtgcg tttttggttt tttgaaagtg cgtcatcaga gcgcttttgg
     4321 ttttcaaaag cgctctgaag ttcctatact ttctagctag agaataggaa cttcccgagc
     4381 ggccgcgtgt tacaagcttg actccagcgt aactggactg caatcaactc actggctcac
     4441 cttcacgggt gggcctttct tcggtagaaa atcaaaggat cttcttgaga tccttttttt
     4501 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4561 ccggatcaag agctaccaac tctttttccg aggtaactgg cttcagcaga gcgcagatac
     4621 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4681 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4741 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4801 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4861 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4921 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca ggggtaaacg
     4981 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcat cgatttttgt
     5041 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcagaa aggcccaccc
     5101 gaaggtgagc caggtgatta catttgggcc ctcatcagag gttttcaccg tcatcaccga
     5161 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
     5221 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
     5281 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcacttac caatgcttaa
     5341 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5401 ccgtcgtgta gataactacg atgcgggagg gcttaccatc tggccccagt gctgcaatga
     5461 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggga
     5521 gggccgagcg cagaagtgat cctgcaactt tatccgcctc catccagtct attaattgtt
     5581 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5641 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5701 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5761 gtcctccgat cgttgccaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5821 cactgcataa ttctcttact gtcatgccat ccgtgagatg cttttctgtg actggtgagt
     5881 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5941 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6001 gttcttcggg gcgtaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6061 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6121 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6181 tactcatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta
     6241 gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgccgatca agtcaaaagc
     6301 ctccggtcgg aggcttttga ctttctgcta tggaggtcag gtatgattta aatggtcagt
     6361 taga
//
