LOCUS       sequence-300252-        5629 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 5629)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5629)
  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-m4046230
FEATURES             Location/Qualifiers
     source          1..5629
                     /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..1339
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1384..1402)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1753..2556
                     /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      2639..3621
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3699..3728
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3750..4337
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4349..4376)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4400..4591)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4592..5383,5384..5452))
                     /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     5215..5234
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5453..5555)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5556..5587)
                     /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 ctttgctcgt
     1261 gccgagaggt tctgattata aagatcatga tggtgattat aaagatcatg atattgatta
     1321 taaagatgat gatgataaat aaacaggccc cttttccttt gtcgatatca tgtaattagt
     1381 tatgtcacgc ttacattcac gccctcctcc cacatccgct ctaaccgaaa aggaaggagt
     1441 tagacaacct gaagtctagg tccctattta ttttttttaa tagttatgtt agtattaaga
     1501 acgttattta tatttcaaat ttttcttttt tttctgtaca aagcggccgc cgaaccacta
     1561 gtgcactgca gtacacggtt tccttgaaat ttttttgatt cggtaatctc cgaacagaag
     1621 gaagaacgaa ggaaggagca cagacttaga ttggtatata tacgcatatg tagtgttgaa
     1681 gaaacatgaa attgcccagt attcttaacc caactgcaca gaacaaaaac gtgcaggaaa
     1741 cgaagataaa tcatgtcgaa agctacatat aaggaacgtg ctgctactca tcctagtcct
     1801 gttgctgcca agctatttaa tatcatgcac gaaaagcaaa caaacttgtg tgcttcattg
     1861 gatgttcgta ccaccaagga attactggag ttagttgaag cattaggtcc caaaatttgt
     1921 ttactaaaaa cacatgtgga tatcttgact gatttttcca tggagggcac agttaagccg
     1981 ctaaaggcat tatccgccaa gtacaatttt ttactcttcg aggacagaaa atttgctgac
     2041 attggtaata cagtcaaatt gcagtactct gcgggtgtat acagaatagc agaatgggca
     2101 gacattacga atgcacacgg tgtggtgggc ccaggtattg ttagcggttt gaagcaggcg
     2161 gcagaagaag taacaaagga acctagaggc cttttgatgt tagcagaatt gtcatgcaag
     2221 ggctccctat ctactggaga atatactaag ggtactgttg acattgcgaa gagcgacaaa
     2281 gattttgtta tcggctttat tgctcaaaga gacatgggtg gaagagatga aggttacgat
     2341 tggttgatta tgacacccgg tgtgggttta gatgacaagg gagatgcatt gggtcaacag
     2401 tatagaaccg tggatgatgt ggtttctaca ggatctgaca ttattattgt tggaagagga
     2461 ctatttgcaa agggaaggga tgctaaggta gagggtgaac gttacagaaa agcaggctgg
     2521 gaagcatatt tgagaagatg cggccagcaa aactaaaaaa ctgtattata agtaaatgca
     2581 tgtatactaa actcacaaat tagagcttca atttaattat atcagttatt acccgagtaa
     2641 cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac gcgagagcgc taatttttca
     2701 aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgaaag cgctatttta
     2761 ccaacgaaga atctgtgctt catttttgta aaacaaaaat gcaacgcgag agcgctaatt
     2821 tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc gagagcgcta
     2881 ttttaccaac aaagaatcta tacttctttt ttgttctaca aaaatgcatc ccgagagcgc
     2941 tatttttcta acaaagcatc ttagattact ttttttctcc tttgtgcgct ctataatgca
     3001 gtctcttgat aactttttgc actgtaggtc cgttaaggtt agaagaaggc tactttggtg
     3061 tctattttct cttccataaa aaaagcctga ctccacttcc cgcgtttact gattactagc
     3121 gaagctgcgg gtgcattttt tcaagataaa ggcatccccg attatattct ataccgatgt
     3181 ggattgcgca tactttgtga acagaaagtg atagcgttga tgattcttca ttggtcagaa
     3241 aattatgaac ggtttcttct attttgtctc tatatactac gtataggaaa tgtttacatt
     3301 ttcgtattgt tttcgattca ctctatgaat agttcttact acaatttttt tgtctaaaga
     3361 gtaatactag agataaacat aaaaaatgta gaggtcgagt ttagatgcaa gttcaaggag
     3421 cgaaaggtgg atgggtaggt tatataggga tatagcacag agatatatag caaagagata
     3481 cttttgagca atgtttgtgg aagcggtatt cgcaatattt tagtagctcg ttacagtccg
     3541 gtgcgttttt ggttttttga aagtgcgtca tcagagcgct tttggttttc aaaagcgctc
     3601 tgaagttcct atactttcta gctagagaat aggaacttcc cgagcggccg cgtgttacaa
     3661 gcttgactcc agcgtaactg gactgcaatc aactcactgg ctcaccttca cgggtgggcc
     3721 tttcttcggt agaaaatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3781 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3841 ccaactcttt ttccgaggta actggcttca gcagagcgca gataccaaat actgttcttc
     3901 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3961 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4021 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4081 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4141 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4201 gggtcggaac aggagagcgc acgagggagc ttccaggggt aaacgcctgg tatctttata
     4261 gtcctgtcgg gtttcgccac ctctgacttg agcatcgatt tttgtgatgc tcgtcagggg
     4321 ggcggagcct atggaaaaac gccagcaacg cagaaaggcc cacccgaagg tgagccaggt
     4381 gattacattt gggccctcat cagaggtttt caccgtcatc accgaaacgc gcgaggcagc
     4441 tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg
     4501 cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca
     4561 tgttaagggc ggttttttcc tgtttggtca cttaccaatg cttaatcagt gaggcaccta
     4621 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4681 ctacgatgcg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4741 gctcaccggc tccagattta tcagcaataa accagccagc cgggagggcc gagcgcagaa
     4801 gtgatcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4861 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     4921 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4981 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5041 ccagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5101 ttactgtcat gccatccgtg agatgctttt ctgtgactgg tgagtactca accaagtcat
     5161 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5221 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgta
     5281 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5341 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5401 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc attttagctt
     5461 ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat
     5521 tatggtgaaa gttggaacct cttacgtgcc gatcaagtca aaagcctccg gtcggaggct
     5581 tttgactttc tgctatggag gtcaggtatg atttaaatgg tcagttaga
//
