LOCUS       sequence-291473-        6661 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 ORF3a protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6661)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6661)
  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-m4046592
FEATURES             Location/Qualifiers
     source          1..6661
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             1256..1273
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2416..2434)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2785..3588
                     /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      3671..4653
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4731..4760
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4782..5369
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5381..5408)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5432..5623)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5624..6415,6416..6484))
                     /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     6247..6266
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6485..6587)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6588..6619)
                     /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 atggatttgt ttatgagaat cttcacaatt ggaactgtaa ctttgaagca
      481 aggtgaaatc aaggatgcta ctccttcaga ttttgttcgc gctactgcaa cgataccgat
      541 acaagcctca ctccctttcg gatggcttat tgttggcgtt gcacttcttg ctgtttttca
      601 gagcgcttcc aaaatcataa ccctcaaaaa gagatggcaa ctagcactct ccaagggtgt
      661 tcactttgtt tgcaacttgc tgttgttgtt tgtaacagtt tactcacacc ttttgctcgt
      721 tgctgctggc cttgaagccc cttttctcta tctttatgct ttagtctact tcttgcagag
      781 tataaacttt gtaagaataa taatgaggct ttggctttgc tggaaatgcc gttccaaaaa
      841 cccattactt tatgatgcca actattttct ttgctggcat actaattgtt acgactattg
      901 tataccttac aatagtgtaa cttcttcaat tgtcattact tcaggtgatg gcacaacaag
      961 tcctatttct gaacatgact accagattgg tggttatact gaaaaatggg aatctggagt
     1021 aaaagactgt gttgtattac acagttactt cacttcagac tattaccagc tgtactcaac
     1081 tcaattgagt acagacactg gtgttgaaca tgttaccttc ttcatctaca ataaaattgt
     1141 tgatgagcct gaagaacatg tccaaattca cacaatcgac ggttcatccg gagttgttaa
     1201 tccagtaatg gaaccaattt atgatgaacc gacgacgact actagcgtgc ctttgctcgt
     1261 gccgagaggt tctaaaatcg aggaaggtaa gctagtcatt tggataaatg gagataaggg
     1321 ttacaacggt ttggcagaag taggcaagaa atttgaaaag gacacaggta tcaaggtgac
     1381 ggttgaacat ccagacaagt tagaagaaaa attccctcaa gttgctgcta ctggtgatgg
     1441 tcccgacatt attttctggg cccacgatcg ttttggtgga tatgctcagt caggtttgct
     1501 ggctgagatc accccagaca aggccttcca agataaattg tacccattca cttgggatgc
     1561 tgtcagatac aacggtaagc ttatagcata tccaattgct gttgaagcct tatcgttgat
     1621 ctacaataag gacttgttac ctaacccacc aaagacatgg gaagaaattc cagctttgga
     1681 taaagagcta aaggccaagg gaaaaagcgc tttgatgttt aacttacaag aaccttattt
     1741 cacctggcca ttaatcgctg ctgacggtgg ttacgcattt aaatacgaaa acggtaagta
     1801 tgacattaag gatgtcggcg ttgataatgc tggtgcaaag gctggtttga ccttcttggt
     1861 tgatttaata aagaataaac acatgaacgc tgacactgac tactctatcg ccgaagccgc
     1921 tttcaacaag ggtgaaactg ctatgaccat aaatggtcca tgggcctgga gtaacattga
     1981 tacttctaaa gtaaactatg gtgtcactgt tttgcctaca ttcaaaggcc aaccatccaa
     2041 gccctttgtc ggtgttctat ccgctggcat caatgctgct tctccaaaca aggagctagc
     2101 taaagaattt ttggaaaatt acttgttaac tgacgaaggt ttggaggccg ttaacaagga
     2161 caagccattg ggtgctgtcg ctttgaaatc ttacgaagaa gaattggcta aggatcccag
     2221 aattgccgct accatggaga acgcccaaaa gggtgaaatt atgccaaaca tcccacaaat
     2281 gtcagccttc tggtacgcag ttaggactgc cgtgattaat gctgcttccg gtagacaaac
     2341 cgtcgacgaa gcattaaaag atgctcaaac gtaaacaggc cccttttcct ttgtcgatat
     2401 catgtaatta gttatgtcac gcttacattc acgccctcct cccacatccg ctctaaccga
     2461 aaaggaagga gttagacaac ctgaagtcta ggtccctatt tatttttttt aatagttatg
     2521 ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta caaagcggcc
     2581 gccgaaccac tagtgcactg cagtacacgg tttccttgaa atttttttga ttcggtaatc
     2641 tccgaacaga aggaagaacg aaggaaggag cacagactta gattggtata tatacgcata
     2701 tgtagtgttg aagaaacatg aaattgccca gtattcttaa cccaactgca cagaacaaaa
     2761 acgtgcagga aacgaagata aatcatgtcg aaagctacat ataaggaacg tgctgctact
     2821 catcctagtc ctgttgctgc caagctattt aatatcatgc acgaaaagca aacaaacttg
     2881 tgtgcttcat tggatgttcg taccaccaag gaattactgg agttagttga agcattaggt
     2941 cccaaaattt gtttactaaa aacacatgtg gatatcttga ctgatttttc catggagggc
     3001 acagttaagc cgctaaaggc attatccgcc aagtacaatt ttttactctt cgaggacaga
     3061 aaatttgctg acattggtaa tacagtcaaa ttgcagtact ctgcgggtgt atacagaata
     3121 gcagaatggg cagacattac gaatgcacac ggtgtggtgg gcccaggtat tgttagcggt
     3181 ttgaagcagg cggcagaaga agtaacaaag gaacctagag gccttttgat gttagcagaa
     3241 ttgtcatgca agggctccct atctactgga gaatatacta agggtactgt tgacattgcg
     3301 aagagcgaca aagattttgt tatcggcttt attgctcaaa gagacatggg tggaagagat
     3361 gaaggttacg attggttgat tatgacaccc ggtgtgggtt tagatgacaa gggagatgca
     3421 ttgggtcaac agtatagaac cgtggatgat gtggtttcta caggatctga cattattatt
     3481 gttggaagag gactatttgc aaagggaagg gatgctaagg tagagggtga acgttacaga
     3541 aaagcaggct gggaagcata tttgagaaga tgcggccagc aaaactaaaa aactgtatta
     3601 taagtaaatg catgtatact aaactcacaa attagagctt caatttaatt atatcagtta
     3661 ttacccgagt aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc
     3721 gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa
     3781 agcgctattt taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg
     3841 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     3901 gcgagagcgc tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca
     3961 tcccgagagc gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg
     4021 ctctataatg cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag
     4081 gctactttgg tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta
     4141 ctgattacta gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt
     4201 ctataccgat gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt
     4261 cattggtcag aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga
     4321 aatgtttaca ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt
     4381 tttgtctaaa gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc
     4441 aagttcaagg agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat
     4501 agcaaagaga tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct
     4561 cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt catcagagcg cttttggttt
     4621 tcaaaagcgc tctgaagttc ctatactttc tagctagaga ataggaactt cccgagcggc
     4681 cgcgtgttac aagcttgact ccagcgtaac tggactgcaa tcaactcact ggctcacctt
     4741 cacgggtggg cctttcttcg gtagaaaatc aaaggatctt cttgagatcc tttttttctg
     4801 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4861 gatcaagagc taccaactct ttttccgagg taactggctt cagcagagcg cagataccaa
     4921 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4981 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5041 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5101 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5161 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5221 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg gtaaacgcct
     5281 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcatcga tttttgtgat
     5341 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcagaaagg cccacccgaa
     5401 ggtgagccag gtgattacat ttgggccctc atcagaggtt ttcaccgtca tcaccgaaac
     5461 gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg
     5521 cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga
     5581 taaagcgggc catgttaagg gcggtttttt cctgtttggt cacttaccaa tgcttaatca
     5641 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5701 tcgtgtagat aactacgatg cgggagggct taccatctgg ccccagtgct gcaatgatac
     5761 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccgggaggg
     5821 ccgagcgcag aagtgatcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     5881 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     5941 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6001 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6061 ctccgatcgt tgccagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6121 tgcataattc tcttactgtc atgccatccg tgagatgctt ttctgtgact ggtgagtact
     6181 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6241 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6301 cttcggggcg taaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6361 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6421 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6481 tcattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg
     6541 atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaagt caaaagcctc
     6601 cggtcggagg cttttgactt tctgctatgg aggtcaggta tgatttaaat ggtcagttag
     6661 a
//
