LOCUS       sequence-287659-        6012 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 6012)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6012)
  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-m4046496
FEATURES             Location/Qualifiers
     source          1..6012
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..466
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    25..142
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     437..460
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             607..624
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1767..1785)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2136..2939
                     /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      3022..4004
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4082..4111
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4133..4720
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4732..4759)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4783..4974)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4975..5766,5767..5835))
                     /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     5598..5617
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5836..5938)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5939..5970)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgctt gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg
       61 acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg
      121 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat
      181 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta
      241 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag
      301 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt
      361 taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta
      421 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aactataatg
      481 atcgagttat ccttgataga tttctatcta tgttttttgg cattcctgct ttttctcgtg
      541 ttgattatgt tgattatctt ctggttcagt ttagaattgc aagaccataa tgaaacatgc
      601 cacgccctcg tgccgagagg ttctaaaatc gaggaaggta agctagtcat ttggataaat
      661 ggagataagg gttacaacgg tttggcagaa gtaggcaaga aatttgaaaa ggacacaggt
      721 atcaaggtga cggttgaaca tccagacaag ttagaagaaa aattccctca agttgctgct
      781 actggtgatg gtcccgacat tattttctgg gcccacgatc gttttggtgg atatgctcag
      841 tcaggtttgc tggctgagat caccccagac aaggccttcc aagataaatt gtacccattc
      901 acttgggatg ctgtcagata caacggtaag cttatagcat atccaattgc tgttgaagcc
      961 ttatcgttga tctacaataa ggacttgtta cctaacccac caaagacatg ggaagaaatt
     1021 ccagctttgg ataaagagct aaaggccaag ggaaaaagcg ctttgatgtt taacttacaa
     1081 gaaccttatt tcacctggcc attaatcgct gctgacggtg gttacgcatt taaatacgaa
     1141 aacggtaagt atgacattaa ggatgtcggc gttgataatg ctggtgcaaa ggctggtttg
     1201 accttcttgg ttgatttaat aaagaataaa cacatgaacg ctgacactga ctactctatc
     1261 gccgaagccg ctttcaacaa gggtgaaact gctatgacca taaatggtcc atgggcctgg
     1321 agtaacattg atacttctaa agtaaactat ggtgtcactg ttttgcctac attcaaaggc
     1381 caaccatcca agccctttgt cggtgttcta tccgctggca tcaatgctgc ttctccaaac
     1441 aaggagctag ctaaagaatt tttggaaaat tacttgttaa ctgacgaagg tttggaggcc
     1501 gttaacaagg acaagccatt gggtgctgtc gctttgaaat cttacgaaga agaattggct
     1561 aaggatccca gaattgccgc taccatggag aacgcccaaa agggtgaaat tatgccaaac
     1621 atcccacaaa tgtcagcctt ctggtacgca gttaggactg ccgtgattaa tgctgcttcc
     1681 ggtagacaaa ccgtcgacga agcattaaaa gatgctcaaa cgtaaacagg ccccttttcc
     1741 tttgtcgata tcatgtaatt agttatgtca cgcttacatt cacgccctcc tcccacatcc
     1801 gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat ttattttttt
     1861 taatagttat gttagtatta agaacgttat ttatatttca aatttttctt ttttttctgt
     1921 acaaagcggc cgccgaacca ctagtgcact gcagtacacg gtttccttga aatttttttg
     1981 attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt agattggtat
     2041 atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta acccaactgc
     2101 acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca tataaggaac
     2161 gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg cacgaaaagc
     2221 aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg gagttagttg
     2281 aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg actgattttt
     2341 ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat tttttactct
     2401 tcgaggacag aaaatttgct gacattggta atacagtcaa attgcagtac tctgcgggtg
     2461 tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg ggcccaggta
     2521 ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga ggccttttga
     2581 tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact aagggtactg
     2641 ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa agagacatgg
     2701 gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt ttagatgaca
     2761 agggagatgc attgggtcaa cagtatagaa ccgtggatga tgtggtttct acaggatctg
     2821 acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag gtagagggtg
     2881 aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag caaaactaaa
     2941 aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct tcaatttaat
     3001 tatatcagtt attacccgag taacgaagca tctgtgcttc attttgtaga acaaaaatgc
     3061 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa
     3121 tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt gtaaaacaaa
     3181 aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac
     3241 agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct tttttgttct
     3301 acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt actttttttc
     3361 tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag gtccgttaag
     3421 gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc tgactccact
     3481 tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc
     3541 ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa gtgatagcgt
     3601 tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt ctctatatac
     3661 tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg aatagttctt
     3721 actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat gtagaggtcg
     3781 agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag ggatatagca
     3841 cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt attcgcaata
     3901 ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg tcatcagagc
     3961 gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag aataggaact
     4021 tcccgagcgg ccgcgtgtta caagcttgac tccagcgtaa ctggactgca atcaactcac
     4081 tggctcacct tcacgggtgg gcctttcttc ggtagaaaat caaaggatct tcttgagatc
     4141 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4201 tttgtttgcc ggatcaagag ctaccaactc tttttccgag gtaactggct tcagcagagc
     4261 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4321 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4381 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4441 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4501 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4561 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4621 ggtaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcatcg
     4681 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcagaaag
     4741 gcccacccga aggtgagcca ggtgattaca tttgggccct catcagaggt tttcaccgtc
     4801 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc
     4861 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
     4921 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcacttacca
     4981 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5041 ctgactcccc gtcgtgtaga taactacgat gcgggagggc ttaccatctg gccccagtgc
     5101 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5161 agccgggagg gccgagcgca gaagtgatcc tgcaacttta tccgcctcca tccagtctat
     5221 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5281 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5341 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5401 ctccttcggt cctccgatcg ttgccagaag taagttggcc gcagtgttat cactcatggt
     5461 tatggcagca ctgcataatt ctcttactgt catgccatcc gtgagatgct tttctgtgac
     5521 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5581 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5641 tggaaaacgt tcttcggggc gtaaactctc aaggatctta ccgctgttga gatccagttc
     5701 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5761 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5821 atgttgaata ctcattttag cttccttagc tcctgaaaat ctcgataact caaaaaatac
     5881 gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt gccgatcaag
     5941 tcaaaagcct ccggtcggag gcttttgact ttctgctatg gaggtcaggt atgatttaaa
     6001 tggtcagtta ga
//
