LOCUS       sequence-287633-        4876 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 4876)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4876)
  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-m4046569
FEATURES             Location/Qualifiers
     source          1..4876
                     /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             545..562
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             563..586
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(631..649)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1000..1803
                     /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      1886..2868
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      2946..2975
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      2997..3584
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3596..3623)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3647..3838)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3839..4630,4631..4699))
                     /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     4462..4481
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4700..4802)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4803..4834)
                     /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 atgggttaca taaatgtctt tgcattccct ttcaccatct atagtcttct
      481 attatgcaga atgaactccc gaaactacat tgcccaggtt gacgtagtta actttaattt
      541 gactctcgtg ccgagaggtt cttggagcca tccgcagttt gaaaaataaa caggcccctt
      601 ttcctttgtc gatatcatgt aattagttat gtcacgctta cattcacgcc ctcctcccac
      661 atccgctcta accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt
      721 tttttaatag ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt
      781 ctgtacaaag cggccgccga accactagtg cactgcagta cacggtttcc ttgaaatttt
      841 tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg
      901 gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa
      961 ctgcacagaa caaaaacgtg caggaaacga agataaatca tgtcgaaagc tacatataag
     1021 gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa
     1081 aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta
     1141 gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat
     1201 ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta
     1261 ctcttcgagg acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg
     1321 ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca
     1381 ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt
     1441 ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt
     1501 actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac
     1561 atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat
     1621 gacaagggag atgcattggg tcaacagtat agaaccgtgg atgatgtggt ttctacagga
     1681 tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag
     1741 ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac
     1801 taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt
     1861 taattatatc agttattacc cgagtaacga agcatctgtg cttcattttg tagaacaaaa
     1921 atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca
     1981 gaaatgcaac gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa
     2041 caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca
     2101 gaacagaaat gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg
     2161 ttctacaaaa atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt
     2221 tttctccttt gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt
     2281 taaggttaga agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc
     2341 cacttcccgc gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc
     2401 atccccgatt atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata
     2461 gcgttgatga ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat
     2521 atactacgta taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt
     2581 tcttactaca atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag
     2641 gtcgagttta gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat
     2701 agcacagaga tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc
     2761 aatattttag tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcatca
     2821 gagcgctttt ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg
     2881 aacttcccga gcggccgcgt gttacaagct tgactccagc gtaactggac tgcaatcaac
     2941 tcactggctc accttcacgg gtgggccttt cttcggtaga aaatcaaagg atcttcttga
     3001 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3061 gtggtttgtt tgccggatca agagctacca actctttttc cgaggtaact ggcttcagca
     3121 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     3181 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3241 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3301 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3361 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3421 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3481 caggggtaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3541 atcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcag
     3601 aaaggcccac ccgaaggtga gccaggtgat tacatttggg ccctcatcag aggttttcac
     3661 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     3721 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     3781 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactt
     3841 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3901 ttgcctgact ccccgtcgtg tagataacta cgatgcggga gggcttacca tctggcccca
     3961 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4021 agccagccgg gagggccgag cgcagaagtg atcctgcaac tttatccgcc tccatccagt
     4081 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4141 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4201 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4261 ttagctcctt cggtcctccg atcgttgcca gaagtaagtt ggccgcagtg ttatcactca
     4321 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtgaga tgcttttctg
     4381 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4441 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4501 tcattggaaa acgttcttcg gggcgtaaac tctcaaggat cttaccgctg ttgagatcca
     4561 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4621 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4681 ggaaatgttg aatactcatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa
     4741 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat
     4801 caagtcaaaa gcctccggtc ggaggctttt gactttctgc tatggaggtc aggtatgatt
     4861 taaatggtca gttaga
//
