LOCUS       sequence-292476-        5299 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 5299)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5299)
  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-m4046498
FEATURES             Location/Qualifiers
     source          1..5299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        13..420
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             971..988
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             989..1012
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1057..1075)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1426..2229
                     /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      2312..3294
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3372..3401
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3423..4010
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4022..4049)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4073..4264)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4265..5056,5057..5125))
                     /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     4888..4907
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5126..5228)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5229..5260)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 agagcggccg ccccacacac catagcttca aaatgtttct actccttttt tactcttcca
       61 gattttctcg gactccgcgc atcgccgtac cacttcaaaa cacccaagca cagcatacta
      121 aatttcccct ctttcttcct ctagggtgtc gttaattacc cgtactaaag gtttggaaaa
      181 gaaaaaagag accgcctcgt ttctttttct tcgtcgaaaa aggcaataaa aatttttatc
      241 acgtttcttt ttcttgaaaa tttttttttt tgattttttt ctctttcgat gacctcccat
      301 tgatatttaa gttaataaac ggtcttcaat ttctcaagtt tcagtttcat ttttcttgtt
      361 ctattacaac tttttttact tcttgctcat tagaaagaaa gcatagcaat ctaatctaag
      421 ttttaattac aaaatggaga gccttgtccc tggtttcaac gagaaaacac acgtccaact
      481 cagtttgcct gttttacagg ttcgcgacgt gctcgtacgt ggctttggag actccgtgga
      541 ggaggtctta tcagaggcac gtcaacatct taaagatggc acttgtggct tagtagaagt
      601 tgaaaaaggc gttttgcctc aacttgaaca gccctatgtg ttcatcaaac gttcggatgc
      661 tcgaactgca cctcatggtc atgttatggt tgagctggta gcagaactcg aaggcattca
      721 gtacggtcgt agtggtgaga cacttggtgt ccttgtccct catgtgggcg aaataccagt
      781 ggcttaccgc aaggttcttc ttcgtaagaa cggtaataaa ggagctggtg gccatagtta
      841 cggcgccgat ctaaagtcat ttgacttagg cgacgagctt ggcactgatc cttatgaaga
      901 ttttcaagaa aactggaaca ctaaacatag cagtggtgtt acccgtgaac tcatgcgtga
      961 gcttaacgga ctcgtgccga gaggttcttg gagccatccg cagtttgaaa aataaacagg
     1021 ccccttttcc tttgtcgata tcatgtaatt agttatgtca cgcttacatt cacgccctcc
     1081 tcccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat
     1141 ttattttttt taatagttat gttagtatta agaacgttat ttatatttca aatttttctt
     1201 ttttttctgt acaaagcggc cgccgaacca ctagtgcact gcagtacacg gtttccttga
     1261 aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt
     1321 agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta
     1381 acccaactgc acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca
     1441 tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg
     1501 cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg
     1561 gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg
     1621 actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat
     1681 tttttactct tcgaggacag aaaatttgct gacattggta atacagtcaa attgcagtac
     1741 tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg
     1801 ggcccaggta ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga
     1861 ggccttttga tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact
     1921 aagggtactg ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa
     1981 agagacatgg gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt
     2041 ttagatgaca agggagatgc attgggtcaa cagtatagaa ccgtggatga tgtggtttct
     2101 acaggatctg acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag
     2161 gtagagggtg aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag
     2221 caaaactaaa aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct
     2281 tcaatttaat tatatcagtt attacccgag taacgaagca tctgtgcttc attttgtaga
     2341 acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac
     2401 agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt
     2461 gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     2521 tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct
     2581 tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt
     2641 actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag
     2701 gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc
     2761 tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat
     2821 aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa
     2881 gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt
     2941 ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg
     3001 aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat
     3061 gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag
     3121 ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt
     3181 attcgcaata ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg
     3241 tcatcagagc gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag
     3301 aataggaact tcccgagcgg ccgcgtgtta caagcttgac tccagcgtaa ctggactgca
     3361 atcaactcac tggctcacct tcacgggtgg gcctttcttc ggtagaaaat caaaggatct
     3421 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3481 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgag gtaactggct
     3541 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     3601 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3661 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3721 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3781 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3841 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3901 agcttccagg ggtaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3961 ttgagcatcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4021 acgcagaaag gcccacccga aggtgagcca ggtgattaca tttgggccct catcagaggt
     4081 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt
     4141 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa
     4201 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg
     4261 tcacttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4321 ccatagttgc ctgactcccc gtcgtgtaga taactacgat gcgggagggc ttaccatctg
     4381 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4441 taaaccagcc agccgggagg gccgagcgca gaagtgatcc tgcaacttta tccgcctcca
     4501 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4561 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4621 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4681 aagcggttag ctccttcggt cctccgatcg ttgccagaag taagttggcc gcagtgttat
     4741 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtgagatgct
     4801 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4861 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     4921 tgctcatcat tggaaaacgt tcttcggggc gtaaactctc aaggatctta ccgctgttga
     4981 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5041 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5101 cgacacggaa atgttgaata ctcattttag cttccttagc tcctgaaaat ctcgataact
     5161 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt
     5221 gccgatcaag tcaaaagcct ccggtcggag gcttttgact ttctgctatg gaggtcaggt
     5281 atgatttaaa tggtcagtt
//
