LOCUS       sequence-297776-        5587 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 5587)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5587)
  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-m4046287
FEATURES             Location/Qualifiers
     source          1..5587
                     /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..1297
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1342..1360)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1711..2514
                     /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      2597..3579
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3657..3686
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3708..4295
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4307..4334)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4358..4549)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4550..5341,5342..5410))
                     /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     5173..5192
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5411..5513)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5514..5545)
                     /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 tttnnnnttc 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 tcttggagcc atccgcagtt tgaaaaataa acaggcccct tttcctttgt
     1321 cgatatcatg taattagtta tgtcacgctt acattcacgc cctcctccca catccgctct
     1381 aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt ttttttaata
     1441 gttatgttag tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaaa
     1501 gcggccgccg aaccactagt gcactgcagt acacggtttc cttgaaattt ttttgattcg
     1561 gtaatctccg aacagaagga agaacgaagg aaggagcaca gacttagatt ggtatatata
     1621 cgcatatgta gtgttgaaga aacatgaaat tgcccagtat tcttaaccca actgcacaga
     1681 acaaaaacgt gcaggaaacg aagataaatc atgtcgaaag ctacatataa ggaacgtgct
     1741 gctactcatc ctagtcctgt tgctgccaag ctatttaata tcatgcacga aaagcaaaca
     1801 aacttgtgtg cttcattgga tgttcgtacc accaaggaat tactggagtt agttgaagca
     1861 ttaggtccca aaatttgttt actaaaaaca catgtggata tcttgactga tttttccatg
     1921 gagggcacag ttaagccgct aaaggcatta tccgccaagt acaatttttt actcttcgag
     1981 gacagaaaat ttgctgacat tggtaataca gtcaaattgc agtactctgc gggtgtatac
     2041 agaatagcag aatgggcaga cattacgaat gcacacggtg tggtgggccc aggtattgtt
     2101 agcggtttga agcaggcggc agaagaagta acaaaggaac ctagaggcct tttgatgtta
     2161 gcagaattgt catgcaaggg ctccctatct actggagaat atactaaggg tactgttgac
     2221 attgcgaaga gcgacaaaga ttttgttatc ggctttattg ctcaaagaga catgggtgga
     2281 agagatgaag gttacgattg gttgattatg acacccggtg tgggtttaga tgacaaggga
     2341 gatgcattgg gtcaacagta tagaaccgtg gatgatgtgg tttctacagg atctgacatt
     2401 attattgttg gaagaggact atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt
     2461 tacagaaaag caggctggga agcatatttg agaagatgcg gccagcaaaa ctaaaaaact
     2521 gtattataag taaatgcatg tatactaaac tcacaaatta gagcttcaat ttaattatat
     2581 cagttattac ccgagtaacg aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc
     2641 gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa
     2701 cgcgaaagcg ctattttacc aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc
     2761 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
     2821 tgcaacgcga gagcgctatt ttaccaacaa agaatctata cttctttttt gttctacaaa
     2881 aatgcatccc gagagcgcta tttttctaac aaagcatctt agattacttt ttttctcctt
     2941 tgtgcgctct ataatgcagt ctcttgataa ctttttgcac tgtaggtccg ttaaggttag
     3001 aagaaggcta ctttggtgtc tattttctct tccataaaaa aagcctgact ccacttcccg
     3061 cgtttactga ttactagcga agctgcgggt gcattttttc aagataaagg catccccgat
     3121 tatattctat accgatgtgg attgcgcata ctttgtgaac agaaagtgat agcgttgatg
     3181 attcttcatt ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta tatactacgt
     3241 ataggaaatg tttacatttt cgtattgttt tcgattcact ctatgaatag ttcttactac
     3301 aatttttttg tctaaagagt aatactagag ataaacataa aaaatgtaga ggtcgagttt
     3361 agatgcaagt tcaaggagcg aaaggtggat gggtaggtta tatagggata tagcacagag
     3421 atatatagca aagagatact tttgagcaat gtttgtggaa gcggtattcg caatatttta
     3481 gtagctcgtt acagtccggt gcgtttttgg ttttttgaaa gtgcgtcatc agagcgcttt
     3541 tggttttcaa aagcgctctg aagttcctat actttctagc tagagaatag gaacttcccg
     3601 agcggccgcg tgttacaagc ttgactccag cgtaactgga ctgcaatcaa ctcactggct
     3661 caccttcacg ggtgggcctt tcttcggtag aaaatcaaag gatcttcttg agatcctttt
     3721 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3781 ttgccggatc aagagctacc aactcttttt ccgaggtaac tggcttcagc agagcgcaga
     3841 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3901 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3961 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4021 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4081 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4141 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccaggggtaa
     4201 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag catcgatttt
     4261 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgca gaaaggccca
     4321 cccgaaggtg agccaggtga ttacatttgg gccctcatca gaggttttca ccgtcatcac
     4381 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     4441 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     4501 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact taccaatgct
     4561 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4621 tccccgtcgt gtagataact acgatgcggg agggcttacc atctggcccc agtgctgcaa
     4681 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4741 ggagggccga gcgcagaagt gatcctgcaa ctttatccgc ctccatccag tctattaatt
     4801 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4861 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4921 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4981 tcggtcctcc gatcgttgcc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5041 cagcactgca taattctctt actgtcatgc catccgtgag atgcttttct gtgactggtg
     5101 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5161 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5221 aacgttcttc ggggcgtaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5281 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5341 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5401 gaatactcat tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg
     5461 gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaagtcaaa
     5521 agcctccggt cggaggcttt tgactttctg ctatggaggt caggtatgat ttaaatggtc
     5581 agttaga
//
