LOCUS       sequence-287728-        5581 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 5581)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5581)
  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-m4046485
FEATURES             Location/Qualifiers
     source          1..5581
                     /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"
     CDS             1274..1291
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1336..1354)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1705..2508
                     /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      2591..3573
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3651..3680
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3702..4289
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4301..4328)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4352..4543)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4544..5335,5336..5404))
                     /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     5167..5186
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5405..5507)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5508..5539)
                     /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 tctcaccatc atcaccatca ttaaacaggc cccttttcct ttgtcgatat
     1321 catgtaatta gttatgtcac gcttacattc acgccctcct cccacatccg ctctaaccga
     1381 aaaggaagga gttagacaac ctgaagtcta ggtccctatt tatttttttt aatagttatg
     1441 ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta caaagcggcc
     1501 gccgaaccac tagtgcactg cagtacacgg tttccttgaa atttttttga ttcggtaatc
     1561 tccgaacaga aggaagaacg aaggaaggag cacagactta gattggtata tatacgcata
     1621 tgtagtgttg aagaaacatg aaattgccca gtattcttaa cccaactgca cagaacaaaa
     1681 acgtgcagga aacgaagata aatcatgtcg aaagctacat ataaggaacg tgctgctact
     1741 catcctagtc ctgttgctgc caagctattt aatatcatgc acgaaaagca aacaaacttg
     1801 tgtgcttcat tggatgttcg taccaccaag gaattactgg agttagttga agcattaggt
     1861 cccaaaattt gtttactaaa aacacatgtg gatatcttga ctgatttttc catggagggc
     1921 acagttaagc cgctaaaggc attatccgcc aagtacaatt ttttactctt cgaggacaga
     1981 aaatttgctg acattggtaa tacagtcaaa ttgcagtact ctgcgggtgt atacagaata
     2041 gcagaatggg cagacattac gaatgcacac ggtgtggtgg gcccaggtat tgttagcggt
     2101 ttgaagcagg cggcagaaga agtaacaaag gaacctagag gccttttgat gttagcagaa
     2161 ttgtcatgca agggctccct atctactgga gaatatacta agggtactgt tgacattgcg
     2221 aagagcgaca aagattttgt tatcggcttt attgctcaaa gagacatggg tggaagagat
     2281 gaaggttacg attggttgat tatgacaccc ggtgtgggtt tagatgacaa gggagatgca
     2341 ttgggtcaac agtatagaac cgtggatgat gtggtttcta caggatctga cattattatt
     2401 gttggaagag gactatttgc aaagggaagg gatgctaagg tagagggtga acgttacaga
     2461 aaagcaggct gggaagcata tttgagaaga tgcggccagc aaaactaaaa aactgtatta
     2521 taagtaaatg catgtatact aaactcacaa attagagctt caatttaatt atatcagtta
     2581 ttacccgagt aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc
     2641 gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa
     2701 agcgctattt taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg
     2761 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     2821 gcgagagcgc tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca
     2881 tcccgagagc gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg
     2941 ctctataatg cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag
     3001 gctactttgg tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta
     3061 ctgattacta gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt
     3121 ctataccgat gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt
     3181 cattggtcag aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga
     3241 aatgtttaca ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt
     3301 tttgtctaaa gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc
     3361 aagttcaagg agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat
     3421 agcaaagaga tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct
     3481 cgttacagtc cggtgcgttt ttggtttttt gaaagtgcgt catcagagcg cttttggttt
     3541 tcaaaagcgc tctgaagttc ctatactttc tagctagaga ataggaactt cccgagcggc
     3601 cgcgtgttac aagcttgact ccagcgtaac tggactgcaa tcaactcact ggctcacctt
     3661 cacgggtggg cctttcttcg gtagaaaatc aaaggatctt cttgagatcc tttttttctg
     3721 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3781 gatcaagagc taccaactct ttttccgagg taactggctt cagcagagcg cagataccaa
     3841 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3901 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3961 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4021 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4081 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4141 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg gtaaacgcct
     4201 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcatcga tttttgtgat
     4261 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcagaaagg cccacccgaa
     4321 ggtgagccag gtgattacat ttgggccctc atcagaggtt ttcaccgtca tcaccgaaac
     4381 gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg
     4441 cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga
     4501 taaagcgggc catgttaagg gcggtttttt cctgtttggt cacttaccaa tgcttaatca
     4561 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4621 tcgtgtagat aactacgatg cgggagggct taccatctgg ccccagtgct gcaatgatac
     4681 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccgggaggg
     4741 ccgagcgcag aagtgatcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4801 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4861 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4921 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4981 ctccgatcgt tgccagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5041 tgcataattc tcttactgtc atgccatccg tgagatgctt ttctgtgact ggtgagtact
     5101 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5161 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5221 cttcggggcg taaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5281 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5341 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5401 tcattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg
     5461 atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaagt caaaagcctc
     5521 cggtcggagg cttttgactt tctgctatgg aggtcaggta tgatttaaat ggtcagttag
     5581 a
//
