LOCUS       sequence-300434-        5592 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 5592)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5592)
  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-m4046698
FEATURES             Location/Qualifiers
     source          1..5592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(382..1185)
                     /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"
     primer_bind     1536..1554
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(2435..2876)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(2441..2464)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    2759..2876
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     terminator      2943..2974
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        2975..3077
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(3078..3146,3147..3938)
                     /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     complement(3296..3315)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             3939..4130
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      4154..4181
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(4193..4780)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4802..4831)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      join(4909..5592,1..299)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 ctctcgggat gcatttttgt agaacaaaaa agaagtatag attctttgtt ggtaaaatag
       61 cgctctcgcg ttgcatttct gttctgtaaa aatgcagctc agattctttg tttgaaaaat
      121 tagcgctctc gcgttgcatt tttgttttac aaaaatgaag cacagattct tcgttggtaa
      181 aatagcgctt tcgcgttgca tttctgttct gtaaaaatgc agctcagatt ctttgtttga
      241 aaaattagcg ctctcgcgtt gcatttttgt tctacaaaat gaagcacaga tgcttcgtta
      301 ctcgggtaat aactgatata attaaattga agctctaatt tgtgagttta gtatacatgc
      361 atttacttat aatacagttt tttagttttg ctggccgcat cttctcaaat atgcttccca
      421 gcctgctttt ctgtaacgtt caccctctac cttagcatcc cttccctttg caaatagtcc
      481 tcttccaaca ataataatgt cagatcctgt agaaaccaca tcatccacgg ttctatactg
      541 ttgacccaat gcatctccct tgtcatctaa acccacaccg ggtgtcataa tcaaccaatc
      601 gtaaccttca tctcttccac ccatgtctct ttgagcaata aagccgataa caaaatcttt
      661 gtcgctcttc gcaatgtcaa cagtaccctt agtatattct ccagtagata gggagccctt
      721 gcatgacaat tctgctaaca tcaaaaggcc tctaggttcc tttgttactt cttctgccgc
      781 ctgcttcaaa ccgctaacaa tacctgggcc caccacaccg tgtgcattcg taatgtctgc
      841 ccattctgct attctgtata cacccgcaga gtactgcaat ttgactgtat taccaatgtc
      901 agcaaatttt ctgtcctcga agagtaaaaa attgtacttg gcggataatg cctttagcgg
      961 cttaactgtg ccctccatgg aaaaatcagt caagatatcc acatgtgttt ttagtaaaca
     1021 aattttggga cctaatgctt caactaactc cagtaattcc ttggtggtac gaacatccaa
     1081 tgaagcacac aagtttgttt gcttttcgtg catgatatta aatagcttgg cagcaacagg
     1141 actaggatga gtagcagcac gttccttata tgtagctttc gacatgattt atcttcgttt
     1201 cctgcacgtt tttgttctgt gcagttgggt taagaatact gggcaatttc atgtttcttc
     1261 aacactacat atgcgtatat ataccaatct aagtctgtgc tccttccttc gttcttcctt
     1321 ctgttcggag attaccgaat caaaaaaatt tcaaggaaac cgtgtactgc agtgcactag
     1381 tggttcggcg gccgctttgt acagaaaaaa aagaaaaatt tgaaatataa ataacgttct
     1441 taatactaac ataactatta aaaaaaataa atagggacct agacttcagg ttgtctaact
     1501 ccttcctttt cggttagagc ggatgtggga ggagggcgtg aatgtaagcg tgacataact
     1561 aattacatga tatcgacaaa ggaaaagggg cctgtttata agggaacaga ggtcgttgtt
     1621 gttggctcat catagattgg ttccataact gggttaacta caccgctaga gccgtcaata
     1681 gtgtgtattt gaacatgttc ttcaggttca tcgactatct tattgtagat gaaaaatgtg
     1741 acgtgttcca ctccggtgtc agtggacaat tgggtgctat acaactggta gtaatcagaa
     1801 gtaaaatagc tgtgtaacac tacgcaatct ttgacaccag actcccactt ttcagtgtaa
     1861 ccaccaattt ggtagtcatg ttctgaaatt ggacttgtgg taccgtcccc ggaggtgata
     1921 actatagagg aagtaaccga gttatatgga atacaataat cgtaacagtt ggtgtgccaa
     1981 caaaggaagt aattagcgtc gtataacaac gggttcttag aacgacattt ccagcacaac
     2041 cacaatctca taatgatacg aacaaagttg atagattgca ggaaatagac caaagcgtac
     2101 aagtacagga atggagcttc taaaccagcg gcaactagga gcaaatggga ataaacagtg
     2161 acaaacaata acaacagatt acagacgaag tgaacaccct tagaaagcgc caattgccat
     2221 ctctttttta aagtaatgat cttactggcg gattggaaaa cagccaagag agcgactcct
     2281 acaatcaacc aaccaaaagg caagctggct tgaatgggga tagtagcggt tgccctaacg
     2341 aaatctgatg gagtagcatc tttaatctca ccctgcttta gggtcactgt gccgatcgtg
     2401 aatattctca taaataagtc cattatagtt ttttctcctt gacgttaaag tatagaggta
     2461 tattaacaat tttttgttga tacttttatg acatttgaat aagaagtaat acaaactgaa
     2521 aatgttgaaa gtattagtta aagtggttat gcagcttttc catttatata tctgttaata
     2581 gatcaaaaat catcgcttcg ctgattaatt accccagaaa taaggctaaa aaactaatcg
     2641 cattatcatc ctatggttgt taatttgatt cgttaatttg aaggtttgtg gggccaggtt
     2701 actgccaatt tttcctcttc ataaccataa aagctagtat tgtagaatct ttattgttcg
     2761 gagcagtgcg gcgcgaggca catctgcgtt tcaggaacgc gaccggtgaa gacgaggacg
     2821 cacggaggag agtcttccgt cggagggctg tcgcccgctc ggcggcttct aatccgtact
     2881 tcaatatagc aagcggccgc tctaactgac catttaaatc atacctgacc tccatagcag
     2941 aaagtcaaaa gcctccgacc ggaggctttt gacttgatcg gcacgtaaga ggttccaact
     3001 ttcaccataa tgaaataaga tcactaccgg gcgtattttt tgagttatcg agattttcag
     3061 gagctaagga agctaaaatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     3121 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     3181 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     3241 ttgagagttt acgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     3301 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     3361 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctc acggatggca
     3421 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     3481 tacttctggc aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     3541 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     3601 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     3661 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     3721 caggatcact tctgcgctcg gccctcccgg ctggctggtt tattgctgat aaatctggag
     3781 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3841 gcatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3901 tcgctgagat aggtgcctca ctgattaagc attggtaagt gaccaaacag gaaaaaaccg
     3961 cccttaacat ggcccgcttt atcagaagcc agacattaac gcttctggag aaactcaacg
     4021 agctggacgc ggatgaacag gcagacatct gtgaatcgct tcacgaccac gctgatgagc
     4081 tttaccgcag ctgcctcgcg cgtttcggtg atgacggtga aaacctctga tgagggccca
     4141 aatgtaatca cctggctcac cttcgggtgg gcctttctgc gttgctggcg tttttccata
     4201 ggctccgccc ccctgacgag catcacaaaa atcgatgctc aagtcagagg tggcgaaacc
     4261 cgacaggact ataaagatac caggcgttta cccctggaag ctccctcgtg cgctctcctg
     4321 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4381 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4441 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4501 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4561 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4621 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt acctcggaaa
     4681 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4741 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgattttcta
     4801 ccgaagaaag gcccacccgt gaaggtgagc cagtgagttg attgcagtcc agttacgctg
     4861 gagtcaagct tgtaacacgc ggccgctcgg gaagttccta ttctctagct agaaagtata
     4921 ggaacttcag agcgcttttg aaaaccaaaa gcgctctgat gacgcacttt caaaaaacca
     4981 aaaacgcacc ggactgtaac gagctactaa aatattgcga ataccgcttc cacaaacatt
     5041 gctcaaaagt atctctttgc tatatatctc tgtgctatat ccctatataa cctacccatc
     5101 cacctttcgc tccttgaact tgcatctaaa ctcgacctct acatttttta tgtttatctc
     5161 tagtattact ctttagacaa aaaaattgta gtaagaacta ttcatagagt gaatcgaaaa
     5221 caatacgaaa atgtaaacat ttcctatacg tagtatatag agacaaaata gaagaaaccg
     5281 ttcataattt tctgaccaat gaagaatcat caacgctatc actttctgtt cacaaagtat
     5341 gcgcaatcca catcggtata gaatataatc ggggatgcct ttatcttgaa aaaatgcacc
     5401 cgcagcttcg ctagtaatca gtaaacgcgg gaagtggagt caggcttttt ttatggaaga
     5461 gaaaatagac accaaagtag ccttcttcta accttaacgg acctacagtg caaaaagtta
     5521 tcaagagact gcattataga gcgcacaaag gagaaaaaaa gtaatctaag atgctttgtt
     5581 agaaaaatag cg
//
