LOCUS       sequence-287679-        5676 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 5676)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5676)
  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-m4046452
FEATURES             Location/Qualifiers
     source          1..5676
                     /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             1303..1320
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1321..1386
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1431..1449)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1800..2603
                     /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      2686..3668
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3746..3775
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3797..4384
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4396..4423)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4447..4638)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4639..5430,5431..5499))
                     /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     5262..5281
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5500..5602)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5603..5634)
                     /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 gacttattta tgagaatatt cacgatcggc acagtgaccc taaagcaggg tgagattaaa
      541 gatgctactc catcagattt cgttagggca accgctacta tccccattca agccagcttg
      601 ccttttggtt ggttgattgt aggagtcgct ctcttggctg ttttccaatc cgccagtaag
      661 atcattactt taaaaaagag atggcaattg gcgctttcta agggtgttca cttcgtctgt
      721 aatctgttgt tattgtttgt cactgtttat tcccatttgc tcctagttgc cgctggttta
      781 gaagctccat tcctgtactt gtacgctttg gtctatttcc tgcaatctat caactttgtt
      841 cgtatcatta tgagattgtg gttgtgctgg aaatgtcgtt ctaagaaccc gttgttatac
      901 gacgctaatt acttcctttg ttggcacacc aactgttacg attattgtat tccatataac
      961 tcggttactt cctctatagt tatcacctcc ggggacggta ccacaagtcc aatttcagaa
     1021 catgactacc aaattggtgg ttacactgaa aagtgggagt ctggtgtcaa agattgcgta
     1081 gtgttacaca gctattttac ttctgattac taccagttgt atagcaccca attgtccact
     1141 gacaccggag tggaacacgt cacatttttc atctacaata agatagtcga tgaacctgaa
     1201 gaacatgttc aaatacacac tattgacggc tctagcggtg tagttaaccc agttatggaa
     1261 ccaatctatg atgagccaac aacaacgacc tctgttccct tactcgtgcc gagaggttct
     1321 gattataaag atcatgatgg tgattataaa gatcatgata ttgattataa agatgatgat
     1381 gataaataaa caggcccctt ttcctttgtc gatatcatgt aattagttat gtcacgctta
     1441 cattcacgcc ctcctcccac atccgctcta accgaaaagg aaggagttag acaacctgaa
     1501 gtctaggtcc ctatttattt tttttaatag ttatgttagt attaagaacg ttatttatat
     1561 ttcaaatttt tctttttttt ctgtacaaag cggccgccga accactagtg cactgcagta
     1621 cacggtttcc ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga
     1681 aggagcacag acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt
     1741 gcccagtatt cttaacccaa ctgcacagaa caaaaacgtg caggaaacga agataaatca
     1801 tgtcgaaagc tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc
     1861 tatttaatat catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca
     1921 ccaaggaatt actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac
     1981 atgtggatat cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat
     2041 ccgccaagta caatttttta ctcttcgagg acagaaaatt tgctgacatt ggtaatacag
     2101 tcaaattgca gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg
     2161 cacacggtgt ggtgggccca ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa
     2221 caaaggaacc tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctatcta
     2281 ctggagaata tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg
     2341 gctttattgc tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga
     2401 cacccggtgt gggtttagat gacaagggag atgcattggg tcaacagtat agaaccgtgg
     2461 atgatgtggt ttctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg
     2521 gaagggatgc taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga
     2581 gaagatgcgg ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact
     2641 cacaaattag agcttcaatt taattatatc agttattacc cgagtaacga agcatctgtg
     2701 cttcattttg tagaacaaaa atgcaacgcg agagcgctaa tttttcaaac aaagaatctg
     2761 agctgcattt ttacagaaca gaaatgcaac gcgaaagcgc tattttacca acgaagaatc
     2821 tgtgcttcat ttttgtaaaa caaaaatgca acgcgagagc gctaattttt caaacaaaga
     2881 atctgagctg catttttaca gaacagaaat gcaacgcgag agcgctattt taccaacaaa
     2941 gaatctatac ttcttttttg ttctacaaaa atgcatcccg agagcgctat ttttctaaca
     3001 aagcatctta gattactttt tttctccttt gtgcgctcta taatgcagtc tcttgataac
     3061 tttttgcact gtaggtccgt taaggttaga agaaggctac tttggtgtct attttctctt
     3121 ccataaaaaa agcctgactc cacttcccgc gtttactgat tactagcgaa gctgcgggtg
     3181 cattttttca agataaaggc atccccgatt atattctata ccgatgtgga ttgcgcatac
     3241 tttgtgaaca gaaagtgata gcgttgatga ttcttcattg gtcagaaaat tatgaacggt
     3301 ttcttctatt ttgtctctat atactacgta taggaaatgt ttacattttc gtattgtttt
     3361 cgattcactc tatgaatagt tcttactaca atttttttgt ctaaagagta atactagaga
     3421 taaacataaa aaatgtagag gtcgagttta gatgcaagtt caaggagcga aaggtggatg
     3481 ggtaggttat atagggatat agcacagaga tatatagcaa agagatactt ttgagcaatg
     3541 tttgtggaag cggtattcgc aatattttag tagctcgtta cagtccggtg cgtttttggt
     3601 tttttgaaag tgcgtcatca gagcgctttt ggttttcaaa agcgctctga agttcctata
     3661 ctttctagct agagaatagg aacttcccga gcggccgcgt gttacaagct tgactccagc
     3721 gtaactggac tgcaatcaac tcactggctc accttcacgg gtgggccttt cttcggtaga
     3781 aaatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3841 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3901 cgaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3961 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4021 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4081 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4141 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4201 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4261 agagcgcacg agggagcttc caggggtaaa cgcctggtat ctttatagtc ctgtcgggtt
     4321 tcgccacctc tgacttgagc atcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4381 gaaaaacgcc agcaacgcag aaaggcccac ccgaaggtga gccaggtgat tacatttggg
     4441 ccctcatcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc
     4501 atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt
     4561 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt
     4621 tttttcctgt ttggtcactt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4681 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatgcggga
     4741 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4801 agatttatca gcaataaacc agccagccgg gagggccgag cgcagaagtg atcctgcaac
     4861 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4921 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4981 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5041 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgcca gaagtaagtt
     5101 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5161 atccgtgaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5221 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5281 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgtaaac tctcaaggat
     5341 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5401 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5461 aaagggaata agggcgacac ggaaatgttg aatactcatt ttagcttcct tagctcctga
     5521 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt
     5581 ggaacctctt acgtgccgat caagtcaaaa gcctccggtc ggaggctttt gactttctgc
     5641 tatggaggtc aggtatgatt taaatggtca gttaga
//
