LOCUS       sequence-297634-        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-m4046330
FEATURES             Location/Qualifiers
     source          1..5997
                     /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             592..609
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1752..1770)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2121..2924
                     /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      3007..3989
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4067..4096
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4118..4705
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(4717..4744)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(4768..4959)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4960..5751,5752..5820))
                     /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     5583..5602
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5821..5923)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5924..5955)
                     /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 ggttacataa atgtctttgc attccctttc accatctata gtcttctatt atgcagaatg
      541 aactcccgaa actacattgc ccaggttgac gtagttaact ttaatttgac tctcgtgccg
      601 agaggttcta aaatcgagga aggtaagcta gtcatttgga taaatggaga taagggttac
      661 aacggtttgg cagaagtagg caagaaattt gaaaaggaca caggtatcaa ggtgacggtt
      721 gaacatccag acaagttaga agaaaaattc cctcaagttg ctgctactgg tgatggtccc
      781 gacattattt tctgggccca cgatcgtttt ggtggatatg ctcagtcagg tttgctggct
      841 gagatcaccc cagacaaggc cttccaagat aaattgtacc cattcacttg ggatgctgtc
      901 agatacaacg gtaagcttat agcatatcca attgctgttg aagccttatc gttgatctac
      961 aataaggact tgttacctaa cccaccaaag acatgggaag aaattccagc tttggataaa
     1021 gagctaaagg ccaagggaaa aagcgctttg atgtttaact tacaagaacc ttatttcacc
     1081 tggccattaa tcgctgctga cggtggttac gcatttaaat acgaaaacgg taagtatgac
     1141 attaaggatg tcggcgttga taatgctggt gcaaaggctg gtttgacctt cttggttgat
     1201 ttaataaaga ataaacacat gaacgctgac actgactact ctatcgccga agccgctttc
     1261 aacaagggtg aaactgctat gaccataaat ggtccatggg cctggagtaa cattgatact
     1321 tctaaagtaa actatggtgt cactgttttg cctacattca aaggccaacc atccaagccc
     1381 tttgtcggtg ttctatccgc tggcatcaat gctgcttctc caaacaagga gctagctaaa
     1441 gaatttttgg aaaattactt gttaactgac gaaggtttgg aggccgttaa caaggacaag
     1501 ccattgggtg ctgtcgcttt gaaatcttac gaagaagaat tggctaagga tcccagaatt
     1561 gccgctacca tggagaacgc ccaaaagggt gaaattatgc caaacatccc acaaatgtca
     1621 gccttctggt acgcagttag gactgccgtg attaatgctg cttccggtag acaaaccgtc
     1681 gacgaagcat taaaagatgc tcaaacgtaa acaggcccct tttcctttgt cgatatcatg
     1741 taattagtta tgtcacgctt acattcacgc cctcctccca catccgctct aaccgaaaag
     1801 gaaggagtta gacaacctga agtctaggtc cctatttatt ttttttaata gttatgttag
     1861 tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaaa gcggccgccg
     1921 aaccactagt gcactgcagt acacggtttc cttgaaattt ttttgattcg gtaatctccg
     1981 aacagaagga agaacgaagg aaggagcaca gacttagatt ggtatatata cgcatatgta
     2041 gtgttgaaga aacatgaaat tgcccagtat tcttaaccca actgcacaga acaaaaacgt
     2101 gcaggaaacg aagataaatc atgtcgaaag ctacatataa ggaacgtgct gctactcatc
     2161 ctagtcctgt tgctgccaag ctatttaata tcatgcacga aaagcaaaca aacttgtgtg
     2221 cttcattgga tgttcgtacc accaaggaat tactggagtt agttgaagca ttaggtccca
     2281 aaatttgttt actaaaaaca catgtggata tcttgactga tttttccatg gagggcacag
     2341 ttaagccgct aaaggcatta tccgccaagt acaatttttt actcttcgag gacagaaaat
     2401 ttgctgacat tggtaataca gtcaaattgc agtactctgc gggtgtatac agaatagcag
     2461 aatgggcaga cattacgaat gcacacggtg tggtgggccc aggtattgtt agcggtttga
     2521 agcaggcggc agaagaagta acaaaggaac ctagaggcct tttgatgtta gcagaattgt
     2581 catgcaaggg ctccctatct actggagaat atactaaggg tactgttgac attgcgaaga
     2641 gcgacaaaga ttttgttatc ggctttattg ctcaaagaga catgggtgga agagatgaag
     2701 gttacgattg gttgattatg acacccggtg tgggtttaga tgacaaggga gatgcattgg
     2761 gtcaacagta tagaaccgtg gatgatgtgg tttctacagg atctgacatt attattgttg
     2821 gaagaggact atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag
     2881 caggctggga agcatatttg agaagatgcg gccagcaaaa ctaaaaaact gtattataag
     2941 taaatgcatg tatactaaac tcacaaatta gagcttcaat ttaattatat cagttattac
     3001 ccgagtaacg aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta
     3061 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg
     3121 ctattttacc aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag
     3181 cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga
     3241 gagcgctatt ttaccaacaa agaatctata cttctttttt gttctacaaa aatgcatccc
     3301 gagagcgcta tttttctaac aaagcatctt agattacttt ttttctcctt tgtgcgctct
     3361 ataatgcagt ctcttgataa ctttttgcac tgtaggtccg ttaaggttag aagaaggcta
     3421 ctttggtgtc tattttctct tccataaaaa aagcctgact ccacttcccg cgtttactga
     3481 ttactagcga agctgcgggt gcattttttc aagataaagg catccccgat tatattctat
     3541 accgatgtgg attgcgcata ctttgtgaac agaaagtgat agcgttgatg attcttcatt
     3601 ggtcagaaaa ttatgaacgg tttcttctat tttgtctcta tatactacgt ataggaaatg
     3661 tttacatttt cgtattgttt tcgattcact ctatgaatag ttcttactac aatttttttg
     3721 tctaaagagt aatactagag ataaacataa aaaatgtaga ggtcgagttt agatgcaagt
     3781 tcaaggagcg aaaggtggat gggtaggtta tatagggata tagcacagag atatatagca
     3841 aagagatact tttgagcaat gtttgtggaa gcggtattcg caatatttta gtagctcgtt
     3901 acagtccggt gcgtttttgg ttttttgaaa gtgcgtcatc agagcgcttt tggttttcaa
     3961 aagcgctctg aagttcctat actttctagc tagagaatag gaacttcccg agcggccgcg
     4021 tgttacaagc ttgactccag cgtaactgga ctgcaatcaa ctcactggct caccttcacg
     4081 ggtgggcctt tcttcggtag aaaatcaaag gatcttcttg agatcctttt tttctgcgcg
     4141 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4201 aagagctacc aactcttttt ccgaggtaac tggcttcagc agagcgcaga taccaaatac
     4261 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     4321 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     4381 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     4441 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     4501 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     4561 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccaggggtaa acgcctggta
     4621 tctttatagt cctgtcgggt ttcgccacct ctgacttgag catcgatttt tgtgatgctc
     4681 gtcagggggg cggagcctat ggaaaaacgc cagcaacgca gaaaggccca cccgaaggtg
     4741 agccaggtga ttacatttgg gccctcatca gaggttttca ccgtcatcac cgaaacgcgc
     4801 gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg
     4861 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
     4921 gcgggccatg ttaagggcgg ttttttcctg tttggtcact taccaatgct taatcagtga
     4981 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5041 gtagataact acgatgcggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5101 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg ggagggccga
     5161 gcgcagaagt gatcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5221 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5281 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5341 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5401 gatcgttgcc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5461 taattctctt actgtcatgc catccgtgag atgcttttct gtgactggtg agtactcaac
     5521 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5581 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5641 ggggcgtaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5701 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5761 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5821 tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct
     5881 tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaagtcaaa agcctccggt
     5941 cggaggcttt tgactttctg ctatggaggt caggtatgat ttaaatggtc agttaga
//
