LOCUS       sequence-337479-        5997 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 ORF10 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5997)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5997)
  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-m4046534
FEATURES             Location/Qualifiers
     source          1..5997
                     /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             568..585
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1728..1746)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2097..2900
                     /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      2983..3965
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4043..4072
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4094..4681
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4693..4720)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4744..4935)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4936..5727,5728..5796))
                     /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     5559..5578
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5797..5899)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5900..5931)
                     /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 ataatgggct atataaacgt tttcgctttt
      481 ccgtttacga tatatagtct actcttgtgc agaatgaatt ctcgtaacta catagcacaa
      541 gtagatgtag ttaactttaa tctcacactc gtgccgagag gttctaaaat cgaggaaggt
      601 aagctagtca tttggataaa tggagataag ggttacaacg gtttggcaga agtaggcaag
      661 aaatttgaaa aggacacagg tatcaaggtg acggttgaac atccagacaa gttagaagaa
      721 aaattccctc aagttgctgc tactggtgat ggtcccgaca ttattttctg ggcccacgat
      781 cgttttggtg gatatgctca gtcaggtttg ctggctgaga tcaccccaga caaggccttc
      841 caagataaat tgtacccatt cacttgggat gctgtcagat acaacggtaa gcttatagca
      901 tatccaattg ctgttgaagc cttatcgttg atctacaata aggacttgtt acctaaccca
      961 ccaaagacat gggaagaaat tccagctttg gataaagagc taaaggccaa gggaaaaagc
     1021 gctttgatgt ttaacttaca agaaccttat ttcacctggc cattaatcgc tgctgacggt
     1081 ggttacgcat ttaaatacga aaacggtaag tatgacatta aggatgtcgg cgttgataat
     1141 gctggtgcaa aggctggttt gaccttcttg gttgatttaa taaagaataa acacatgaac
     1201 gctgacactg actactctat cgccgaagcc gctttcaaca agggtgaaac tgctatgacc
     1261 ataaatggtc catgggcctg gagtaacatt gatacttcta aagtaaacta tggtgtcact
     1321 gttttgccta cattcaaagg ccaaccatcc aagccctttg tcggtgttct atccgctggc
     1381 atcaatgctg cttctccaaa caaggagcta gctaaagaat ttttggaaaa ttacttgtta
     1441 actgacgaag gtttggaggc cgttaacaag gacaagccat tgggtgctgt cgctttgaaa
     1501 tcttacgaag aagaattggc taaggatccc agaattgccg ctaccatgga gaacgcccaa
     1561 aagggtgaaa ttatgccaaa catcccacaa atgtcagcct tctggtacgc agttaggact
     1621 gccgtgatta atgctgcttc cggtagacaa accgtcgacg aagcattaaa agatgctcaa
     1681 acgtaaacag gccccttttc ctttgtcgat atcatgtaat tagttatgtc acgcttacat
     1741 tcacgccctc ctcccacatc cgctctaacc gaaaaggaag gagttagaca acctgaagtc
     1801 taggtcccta tttatttttt ttaatagtta tgttagtatt aagaacgtta tttatatttc
     1861 aaatttttct tttttttctg tacaaagcgg ccgccgaacc actagtgcac tgcagtacac
     1921 ggtttccttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg
     1981 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc
     2041 cagtattctt aacccaactg cacagaacaa aaacgtgcag gaaacgaaga taaatcatgt
     2101 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat
     2161 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca
     2221 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg
     2281 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg
     2341 ccaagtacaa ttttttactc ttcgaggaca gaaaatttgc tgacattggt aatacagtca
     2401 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac
     2461 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa
     2521 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg
     2581 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct
     2641 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac
     2701 ccggtgtggg tttagatgac aagggagatg cattgggtca acagtataga accgtggatg
     2761 atgtggtttc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa
     2821 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa
     2881 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac
     2941 aaattagagc ttcaatttaa ttatatcagt tattacccga gtaacgaagc atctgtgctt
     3001 cattttgtag aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc
     3061 tgcattttta cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt
     3121 gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc
     3181 tgagctgcat ttttacagaa cagaaatgca acgcgagagc gctattttac caacaaagaa
     3241 tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt tctaacaaag
     3301 catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct tgataacttt
     3361 ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt ttctcttcca
     3421 taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct gcgggtgcat
     3481 tttttcaaga taaaggcatc cccgattata ttctataccg atgtggattg cgcatacttt
     3541 gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat gaacggtttc
     3601 ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta ttgttttcga
     3661 ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata ctagagataa
     3721 acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag gtggatgggt
     3781 aggttatata gggatatagc acagagatat atagcaaaga gatacttttg agcaatgttt
     3841 gtggaagcgg tattcgcaat attttagtag ctcgttacag tccggtgcgt ttttggtttt
     3901 ttgaaagtgc gtcatcagag cgcttttggt tttcaaaagc gctctgaagt tcctatactt
     3961 tctagctaga gaataggaac ttcccgagcg gccgcgtgtt acaagcttga ctccagcgta
     4021 actggactgc aatcaactca ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa
     4081 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4141 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4201 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4261 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4321 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4381 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4441 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4501 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4561 gcgcacgagg gagcttccag gggtaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4621 ccacctctga cttgagcatc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4681 aaacgccagc aacgcagaaa ggcccacccg aaggtgagcc aggtgattac atttgggccc
     4741 tcatcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
     4801 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
     4861 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
     4921 ttcctgtttg gtcacttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4981 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tgcgggaggg
     5041 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5101 tttatcagca ataaaccagc cagccgggag ggccgagcgc agaagtgatc ctgcaacttt
     5161 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5221 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5281 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5341 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgccagaa gtaagttggc
     5401 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5461 cgtgagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5521 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5581 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgtaaactct caaggatctt
     5641 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5701 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5761 gggaataagg gcgacacgga aatgttgaat actcatttta gcttccttag ctcctgaaaa
     5821 tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga
     5881 acctcttacg tgccgatcaa gtcaaaagcc tccggtcgga ggcttttgac tttctgctat
     5941 ggaggtcagg tatgatttaa atggtcagtt agagcggccg cttgctatat tgaagta
//
