LOCUS       sequence-291532-        5965 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 ORF7b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5965)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5965)
  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-m4046189
FEATURES             Location/Qualifiers
     source          1..5965
                     /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             560..577
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1720..1738)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2089..2892
                     /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      2975..3957
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4035..4064
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4086..4673
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4685..4712)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4736..4927)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4928..5719,5720..5788))
                     /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     5551..5570
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5789..5891)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5892..5923)
                     /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 atgatcgagt tatccttgat agatttctat ctatgttttt tggcattcct
      481 gctttttctc gtgttgatta tgttgattat cttctggttc agtttagaat tgcaagacca
      541 taatgaaaca tgccacgccc tcgtgccgag aggttctaaa atcgaggaag gtaagctagt
      601 catttggata aatggagata agggttacaa cggtttggca gaagtaggca agaaatttga
      661 aaaggacaca ggtatcaagg tgacggttga acatccagac aagttagaag aaaaattccc
      721 tcaagttgct gctactggtg atggtcccga cattattttc tgggcccacg atcgttttgg
      781 tggatatgct cagtcaggtt tgctggctga gatcacccca gacaaggcct tccaagataa
      841 attgtaccca ttcacttggg atgctgtcag atacaacggt aagcttatag catatccaat
      901 tgctgttgaa gccttatcgt tgatctacaa taaggacttg ttacctaacc caccaaagac
      961 atgggaagaa attccagctt tggataaaga gctaaaggcc aagggaaaaa gcgctttgat
     1021 gtttaactta caagaacctt atttcacctg gccattaatc gctgctgacg gtggttacgc
     1081 atttaaatac gaaaacggta agtatgacat taaggatgtc ggcgttgata atgctggtgc
     1141 aaaggctggt ttgaccttct tggttgattt aataaagaat aaacacatga acgctgacac
     1201 tgactactct atcgccgaag ccgctttcaa caagggtgaa actgctatga ccataaatgg
     1261 tccatgggcc tggagtaaca ttgatacttc taaagtaaac tatggtgtca ctgttttgcc
     1321 tacattcaaa ggccaaccat ccaagccctt tgtcggtgtt ctatccgctg gcatcaatgc
     1381 tgcttctcca aacaaggagc tagctaaaga atttttggaa aattacttgt taactgacga
     1441 aggtttggag gccgttaaca aggacaagcc attgggtgct gtcgctttga aatcttacga
     1501 agaagaattg gctaaggatc ccagaattgc cgctaccatg gagaacgccc aaaagggtga
     1561 aattatgcca aacatcccac aaatgtcagc cttctggtac gcagttagga ctgccgtgat
     1621 taatgctgct tccggtagac aaaccgtcga cgaagcatta aaagatgctc aaacgtaaac
     1681 aggccccttt tcctttgtcg atatcatgta attagttatg tcacgcttac attcacgccc
     1741 tcctcccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc
     1801 tatttatttt ttttaatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt
     1861 cttttttttc tgtacaaagc ggccgccgaa ccactagtgc actgcagtac acggtttcct
     1921 tgaaattttt ttgattcggt aatctccgaa cagaaggaag aacgaaggaa ggagcacaga
     1981 cttagattgg tatatatacg catatgtagt gttgaagaaa catgaaattg cccagtattc
     2041 ttaacccaac tgcacagaac aaaaacgtgc aggaaacgaa gataaatcat gtcgaaagct
     2101 acatataagg aacgtgctgc tactcatcct agtcctgttg ctgccaagct atttaatatc
     2161 atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg ttcgtaccac caaggaatta
     2221 ctggagttag ttgaagcatt aggtcccaaa atttgtttac taaaaacaca tgtggatatc
     2281 ttgactgatt tttccatgga gggcacagtt aagccgctaa aggcattatc cgccaagtac
     2341 aattttttac tcttcgagga cagaaaattt gctgacattg gtaatacagt caaattgcag
     2401 tactctgcgg gtgtatacag aatagcagaa tgggcagaca ttacgaatgc acacggtgtg
     2461 gtgggcccag gtattgttag cggtttgaag caggcggcag aagaagtaac aaaggaacct
     2521 agaggccttt tgatgttagc agaattgtca tgcaagggct ccctatctac tggagaatat
     2581 actaagggta ctgttgacat tgcgaagagc gacaaagatt ttgttatcgg ctttattgct
     2641 caaagagaca tgggtggaag agatgaaggt tacgattggt tgattatgac acccggtgtg
     2701 ggtttagatg acaagggaga tgcattgggt caacagtata gaaccgtgga tgatgtggtt
     2761 tctacaggat ctgacattat tattgttgga agaggactat ttgcaaaggg aagggatgct
     2821 aaggtagagg gtgaacgtta cagaaaagca ggctgggaag catatttgag aagatgcggc
     2881 cagcaaaact aaaaaactgt attataagta aatgcatgta tactaaactc acaaattaga
     2941 gcttcaattt aattatatca gttattaccc gagtaacgaa gcatctgtgc ttcattttgt
     3001 agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt
     3061 tacagaacag aaatgcaacg cgaaagcgct attttaccaa cgaagaatct gtgcttcatt
     3121 tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc
     3181 atttttacag aacagaaatg caacgcgaga gcgctatttt accaacaaag aatctatact
     3241 tcttttttgt tctacaaaaa tgcatcccga gagcgctatt tttctaacaa agcatcttag
     3301 attacttttt ttctcctttg tgcgctctat aatgcagtct cttgataact ttttgcactg
     3361 taggtccgtt aaggttagaa gaaggctact ttggtgtcta ttttctcttc cataaaaaaa
     3421 gcctgactcc acttcccgcg tttactgatt actagcgaag ctgcgggtgc attttttcaa
     3481 gataaaggca tccccgatta tattctatac cgatgtggat tgcgcatact ttgtgaacag
     3541 aaagtgatag cgttgatgat tcttcattgg tcagaaaatt atgaacggtt tcttctattt
     3601 tgtctctata tactacgtat aggaaatgtt tacattttcg tattgttttc gattcactct
     3661 atgaatagtt cttactacaa tttttttgtc taaagagtaa tactagagat aaacataaaa
     3721 aatgtagagg tcgagtttag atgcaagttc aaggagcgaa aggtggatgg gtaggttata
     3781 tagggatata gcacagagat atatagcaaa gagatacttt tgagcaatgt ttgtggaagc
     3841 ggtattcgca atattttagt agctcgttac agtccggtgc gtttttggtt ttttgaaagt
     3901 gcgtcatcag agcgcttttg gttttcaaaa gcgctctgaa gttcctatac tttctagcta
     3961 gagaatagga acttcccgag cggccgcgtg ttacaagctt gactccagcg taactggact
     4021 gcaatcaact cactggctca ccttcacggg tgggcctttc ttcggtagaa aatcaaagga
     4081 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4141 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaggtaactg
     4201 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     4261 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4321 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4381 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4441 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4501 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4561 gggagcttcc aggggtaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4621 gacttgagca tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4681 gcaacgcaga aaggcccacc cgaaggtgag ccaggtgatt acatttgggc cctcatcaga
     4741 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     4801 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     4861 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     4921 tggtcactta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4981 catccatagt tgcctgactc cccgtcgtgt agataactac gatgcgggag ggcttaccat
     5041 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5101 caataaacca gccagccggg agggccgagc gcagaagtga tcctgcaact ttatccgcct
     5161 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5221 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5281 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5341 aaaaagcggt tagctccttc ggtcctccga tcgttgccag aagtaagttg gccgcagtgt
     5401 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtgagat
     5461 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5521 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5581 aagtgctcat cattggaaaa cgttcttcgg ggcgtaaact ctcaaggatc ttaccgctgt
     5641 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5701 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5761 gggcgacacg gaaatgttga atactcattt tagcttcctt agctcctgaa aatctcgata
     5821 actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta
     5881 cgtgccgatc aagtcaaaag cctccggtcg gaggcttttg actttctgct atggaggtca
     5941 ggtatgattt aaatggtcag ttaga
//
