LOCUS       sequence-384616-        5634 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 5634)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5634)
  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-m4046373
FEATURES             Location/Qualifiers
     source          1..5634
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..442
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    1..118
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     413..436
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             1279..1296
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1297..1320
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1365..1383)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1734..2537
                     /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      2620..3602
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3680..3709
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3731..4318
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4330..4357)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4381..4572)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4573..5364,5365..5433))
                     /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     5196..5215
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5434..5536)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5537..5568)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 cggattagaa gccgccgagc gggcgacagc cctccgacgg aagactctcc tccgtgcgtc
       61 ctcgtcttca ccggtcgcgt tcctgaaacg cagatgtgcc tcgcgccgca ctgctccgaa
      121 caataaagat tctacaatac tagcttttat ggttatgaag aggaaaaatt ggcagtaacc
      181 tggccccaca aaccttcaaa ttaacgaatc aaattaacaa ccataggatg ataatgcgat
      241 tagtttttta gccttatttc tggggtaatt aatcagcgaa gcgatgattt ttgatctatt
      301 aacagatata taaatggaaa agctgcataa ccactttaac taatactttc aacattttca
      361 gtttgtatta cttcttattc aaatgtcata aaagtatcaa caaaaaattg ttaatatacc
      421 tctatacttt aacgtcaagg agaaaaaact ataatggact tatttatgag aatattcacg
      481 atcggcacag tgaccctaaa gcagggtgag attaaagatg ctactccatc agatttcgtt
      541 agggcaaccg ctactatccc cattcaagcc agcttgcctt ttggttggtt gattgtagga
      601 gtcgctctct tggctgtttt ccaatccgcc agtaagatca ttactttaaa aaagagatgg
      661 caattggcgc tttctaaggg tgttcacttc gtctgtaatc tgttgttatt gtttgtcact
      721 gtttattccc atttgctcct agttgccgct ggtttagaag ctccattcct gtacttgtac
      781 gctttggtct atttcctgca atctatcaac tttgttcgta tcattatgag attgtggttg
      841 tgctggaaat gtcgttctaa gaacccgttg ttatacgacg ctaattactt cctttgttgg
      901 cacaccaact gttacgatta ttgtattcca tataactcgg ttacttcctc tatagttatc
      961 acctccgggg acggtaccac aagtccaatt tcagaacatg actaccaaat tggtggttac
     1021 actgaaaagt gggagtctgg tgtcaaagat tgcgtagtgt tacacagcta ttttacttct
     1081 gattactacc agttgtatag cacccaattg tccactgaca ccggagtgga acacgtcaca
     1141 tttttcatct acaataagat agtcgatgaa cctgaagaac atgttcaaat acacactatt
     1201 gacggctcta gcggtgtagt taacccagtt atggaaccaa tctatgatga gccaacaaca
     1261 acgacctctg ttcccttact cgtgccgaga ggttcttgga gccatccgca gtttgaaaaa
     1321 taaacaggcc ccttttcctt tgtcgatatc atgtaattag ttatgtcacg cttacattca
     1381 cgccctcctc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag
     1441 gtccctattt atttttttta atagttatgt tagtattaag aacgttattt atatttcaaa
     1501 tttttctttt ttttctgtac aaagcggccg ccgaaccact agtgcactgc agtacacggt
     1561 ttccttgaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga aggaaggagc
     1621 acagacttag attggtatat atacgcatat gtagtgttga agaaacatga aattgcccag
     1681 tattcttaac ccaactgcac agaacaaaaa cgtgcaggaa acgaagataa atcatgtcga
     1741 aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc aagctattta
     1801 atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt accaccaagg
     1861 aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa acacatgtgg
     1921 atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca ttatccgcca
     1981 agtacaattt tttactcttc gaggacagaa aatttgctga cattggtaat acagtcaaat
     2041 tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg aatgcacacg
     2101 gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa gtaacaaagg
     2161 aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta tctactggag
     2221 aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt atcggcttta
     2281 ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt atgacacccg
     2341 gtgtgggttt agatgacaag ggagatgcat tgggtcaaca gtatagaacc gtggatgatg
     2401 tggtttctac aggatctgac attattattg ttggaagagg actatttgca aagggaaggg
     2461 atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat ttgagaagat
     2521 gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta aactcacaaa
     2581 ttagagcttc aatttaatta tatcagttat tacccgagta acgaagcatc tgtgcttcat
     2641 tttgtagaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc
     2701 atttttacag aacagaaatg caacgcgaaa gcgctatttt accaacgaag aatctgtgct
     2761 tcatttttgt aaaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga
     2821 gctgcatttt tacagaacag aaatgcaacg cgagagcgct attttaccaa caaagaatct
     2881 atacttcttt tttgttctac aaaaatgcat cccgagagcg ctatttttct aacaaagcat
     2941 cttagattac tttttttctc ctttgtgcgc tctataatgc nnnctcttga taactttttg
     3001 cactgtaggt cggttaaggt tagaagaagg ctactttggt gtctattttc tcttccataa
     3061 aaaaagcctg actccacttc ccgcgtttac tgattactag cgaagctgcg ggtgcatttt
     3121 ttcaagataa aggcatcccc gattatattc tataccgatg tggattgcgc atactttgtg
     3181 aacagaaagt gatagcgttg atgattcttc attggtcaga aaattatgaa cggtttcttc
     3241 tattttgtct ctatatacta cgtataggaa atgtttacat tttcgtattg ttttcgattc
     3301 actctatgaa tagttcttac tacaattttt ttgtctaaag agtaatacta gagataaaca
     3361 taaaaaatgt agaggtcgag tttagatgca agttcaagga gcgaaaggtg gatgggtagg
     3421 ttatataggg atatagcaca gagatatata gcaaagagat acttttgagc aatgtttgtg
     3481 gaagcggtat tcgcaatatt ttagtagctc gttacagtcc ggtgcgtttt tggttttttg
     3541 aaagtgcgtc atcagagcgc ttttggtttt caaaagcgct ctgaagttcc tatactttct
     3601 agctagagaa taggaacttc ccgagcggcc gcgtgttaca agcttgactc cagcgtaact
     3661 ggactgcaat caactcactg gctcaccttc acgggtgggc ctttcttcgg tagaaaatca
     3721 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3781 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaggt
     3841 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     3901 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3961 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4021 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4081 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4141 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4201 cacgagggag cttccagggg taaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4261 cctctgactt gagcatcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4321 cgccagcaac gcagaaaggc ccacccgaag gtgagccagg tgattacatt tgggccctca
     4381 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc
     4441 gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt
     4501 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc
     4561 ctgtttggtc acttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4621 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatgc gggagggctt
     4681 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4741 atcagcaata aaccagccag ccgggagggc cgagcgcaga agtgatcctg caactttatc
     4801 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4861 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4921 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4981 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gccagaagta agttggccgc
     5041 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5101 gagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5161 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5221 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcgt aaactctcaa ggatcttacc
     5281 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5341 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5401 aataagggcg acacggaaat gttgaatact cattttagct tccttagctc ctgaaaatct
     5461 cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc
     5521 tcttacgtgc cgatcaagtc aaaagcctcc ggtcggaggc ttttgacttt ctgctatgga
     5581 ggtcaggtat gatttaaatg gtcagttaga gcggccgctt gctatattga agta
//
