LOCUS       sequence-298223-        5545 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 5545)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5545)
  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-m4046633
FEATURES             Location/Qualifiers
     source          1..5545
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(1300..1318)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1669..2472
                     /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      2555..3537
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3615..3644
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3666..4253
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4265..4292)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4316..4507)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4508..5299,5300..5368))
                     /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     5131..5150
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5369..5471)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5472..5503)
                     /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 ctttgtaaac
     1261 aggccccttt tcctttgtcg atatcatgta attagttatg tcacgcttac attcacgccc
     1321 tcctcccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc
     1381 tatttatttt ttttaatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt
     1441 cttttttttc tgtacaaagc ggccgccgaa ccactagtgc actgcagtac acggtttcct
     1501 tgaaattttt ttgattcggt aatctccgaa cagaaggaag aacgaaggaa ggagcacaga
     1561 cttagattgg tatatatacg catatgtagt gttgaagaaa catgaaattg cccagtattc
     1621 ttaacccaac tgcacagaac aaaaacgtgc aggaaacgaa gataaatcat gtcgaaagct
     1681 acatataagg aacgtgctgc tactcatcct agtcctgttg ctgccaagct atttaatatc
     1741 atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg ttcgtaccac caaggaatta
     1801 ctggagttag ttgaagcatt aggtcccaaa atttgtttac taaaaacaca tgtggatatc
     1861 ttgactgatt tttccatgga gggcacagtt aagccgctaa aggcattatc cgccaagtac
     1921 aattttttac tcttcgagga cagaaaattt gctgacattg gtaatacagt caaattgcag
     1981 tactctgcgg gtgtatacag aatagcagaa tgggcagaca ttacgaatgc acacggtgtg
     2041 gtgggcccag gtattgttag cggtttgaag caggcggcag aagaagtaac aaaggaacct
     2101 agaggccttt tgatgttagc agaattgtca tgcaagggct ccctatctac tggagaatat
     2161 actaagggta ctgttgacat tgcgaagagc gacaaagatt ttgttatcgg ctttattgct
     2221 caaagagaca tgggtggaag agatgaaggt tacgattggt tgattatgac acccggtgtg
     2281 ggtttagatg acaagggaga tgcattgggt caacagtata gaaccgtgga tgatgtggtt
     2341 tctacaggat ctgacattat tattgttgga agaggactat ttgcaaaggg aagggatgct
     2401 aaggtagagg gtgaacgtta cagaaaagca ggctgggaag catatttgag aagatgcggc
     2461 cagcaaaact aaaaaactgt attataagta aatgcatgta tactaaactc acaaattaga
     2521 gcttcaattt aattatatca gttattaccc gagtaacgaa gcatctgtgc ttcattttgt
     2581 agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt
     2641 tacagaacag aaatgcaacg cgaaagcgct attttaccaa cgaagaatct gtgcttcatt
     2701 tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc
     2761 atttttacag aacagaaatg caacgcgaga gcgctatttt accaacaaag aatctatact
     2821 tcttttttgt tctacaaaaa tgcatcccga gagcgctatt tttctaacaa agcatcttag
     2881 attacttttt ttctcctttg tgcgctctat aatgcagtct cttgataact ttttgcactg
     2941 taggtccgtt aaggttagaa gaaggctact ttggtgtcta ttttctcttc cataaaaaaa
     3001 gcctgactcc acttcccgcg tttactgatt actagcgaag ctgcgggtgc attttttcaa
     3061 gataaaggca tccccgatta tattctatac cgatgtggat tgcgcatact ttgtgaacag
     3121 aaagtgatag cgttgatgat tcttcattgg tcagaaaatt atgaacggtt tcttctattt
     3181 tgtctctata tactacgtat aggaaatgtt tacattttcg tattgttttc gattcactct
     3241 atgaatagtt cttactacaa tttttttgtc taaagagtaa tactagagat aaacataaaa
     3301 aatgtagagg tcgagtttag atgcaagttc aaggagcgaa aggtggatgg gtaggttata
     3361 tagggatata gcacagagat atatagcaaa gagatacttt tgagcaatgt ttgtggaagc
     3421 ggtattcgca atattttagt agctcgttac agtccggtgc gtttttggtt ttttgaaagt
     3481 gcgtcatcag agcgcttttg gttttcaaaa gcgctctgaa gttcctatac tttctagcta
     3541 gagaatagga acttcccgag cggccgcgtg ttacaagctt gactccagcg taactggact
     3601 gcaatcaact cactggctca ccttcacggg tgggcctttc ttcggtagaa aatcaaagga
     3661 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3721 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaggtaactg
     3781 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     3841 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3901 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3961 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4021 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4081 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4141 gggagcttcc aggggtaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4201 gacttgagca tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4261 gcaacgcaga aaggcccacc cgaaggtgag ccaggtgatt acatttgggc cctcatcaga
     4321 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     4381 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     4441 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     4501 tggtcactta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4561 catccatagt tgcctgactc cccgtcgtgt agataactac gatgcgggag ggcttaccat
     4621 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4681 caataaacca gccagccggg agggccgagc gcagaagtga tcctgcaact ttatccgcct
     4741 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4801 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4861 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4921 aaaaagcggt tagctccttc ggtcctccga tcgttgccag aagtaagttg gccgcagtgt
     4981 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtgagat
     5041 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5101 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5161 aagtgctcat cattggaaaa cgttcttcgg ggcgtaaact ctcaaggatc ttaccgctgt
     5221 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5281 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5341 gggcgacacg gaaatgttga atactcattt tagcttcctt agctcctgaa aatctcgata
     5401 actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta
     5461 cgtgccgatc aagtcaaaag cctccggtcg gaggcttttg actttctgct atggaggtca
     5521 ggtatgattt aaatggtcag ttaga
//
