LOCUS       sequence-292483-        5172 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 ORF7a protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5172)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5172)
  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-m4046570
FEATURES             Location/Qualifiers
     source          1..5172
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        28..469
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    28..145
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     440..463
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             844..861
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             862..885
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(930..948)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1299..2102
                     /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      2185..3167
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3245..3274
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3296..3883
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3895..3922)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3946..4137)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4138..4929,4930..4998))
                     /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     4761..4780
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4999..5101)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(5102..5133)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 agagcggccg cttgctatat tgaagtacgg attagaagcc gccgagcggg cgacagccct
       61 ccgacggaag actctcctcc gtgcgtcctc gtcttcaccg gtcgcgttcc tgaaacgcag
      121 atgtgcctcg cgccgcactg ctccgaacaa taaagattct acaatactag cttttatggt
      181 tatgaagagg aaaaattggc agtaacctgg ccccacaaac cttcaaatta acgaatcaaa
      241 ttaacaacca taggatgata atgcgattag ttttttagcc ttatttctgg ggtaattaat
      301 cagcgaagcg atgatttttg atctattaac agatatataa atggaaaagc tgcataacca
      361 ctttaactaa tactttcaac attttcagtt tgtattactt cttattcaaa tgtcataaaa
      421 gtatcaacaa aaaattgtta atatacctct atactttaac gtcaaggaga aaaaactata
      481 atgaaaatta ttcttttctt ggcactgata acactcgcta cttgtgagct ttatcactac
      541 caagagtgtg ttagaggtac aacagtactt ttaaaagaac cttgctcttc tggaacatac
      601 gagggcaatt caccatttca tcctctagct gataacaaat ttgcactgac ttgctttagc
      661 actcaatttg cttttgcttg tcctgacggc gtaaaacacg tctatcagtt acgtgccaga
      721 tcagtttcac ctaaactgtt catcagacaa gaggaagttc aagaacttta ctctccaatt
      781 tttcttattg ttgcggcaat agtgtttata acactttgct tcacactcaa aagaaagaca
      841 gaactcgtgc cgagaggttc ttggagccat ccgcagtttg aaaaataaac aggccccttt
      901 tcctttgtcg atatcatgta attagttatg tcacgcttac attcacgccc tcctcccaca
      961 tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt
     1021 ttttaatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc
     1081 tgtacaaagc ggccgccgaa ccactagtgc actgcagtac acggtttcct tgaaattttt
     1141 ttgattcggt aatctccgaa cagaaggaag aacgaaggaa ggagcacaga cttagattgg
     1201 tatatatacg catatgtagt gttgaagaaa catgaaattg cccagtattc ttaacccaac
     1261 tgcacagaac aaaaacgtgc aggaaacgaa gataaatcat gtcgaaagct acatataagg
     1321 aacgtgctgc tactcatcct agtcctgttg ctgccaagct atttaatatc atgcacgaaa
     1381 agcaaacaaa cttgtgtgct tcattggatg ttcgtaccac caaggaatta ctggagttag
     1441 ttgaagcatt aggtcccaaa atttgtttac taaaaacaca tgtggatatc ttgactgatt
     1501 tttccatgga gggcacagtt aagccgctaa aggcattatc cgccaagtac aattttttac
     1561 tcttcgagga cagaaaattt gctgacattg gtaatacagt caaattgcag tactctgcgg
     1621 gtgtatacag aatagcagaa tgggcagaca ttacgaatgc acacggtgtg gtgggcccag
     1681 gtattgttag cggtttgaag caggcggcag aagaagtaac aaaggaacct agaggccttt
     1741 tgatgttagc agaattgtca tgcaagggct ccctatctac tggagaatat actaagggta
     1801 ctgttgacat tgcgaagagc gacaaagatt ttgttatcgg ctttattgct caaagagaca
     1861 tgggtggaag agatgaaggt tacgattggt tgattatgac acccggtgtg ggtttagatg
     1921 acaagggaga tgcattgggt caacagtata gaaccgtgga tgatgtggtt tctacaggat
     1981 ctgacattat tattgttgga agaggactat ttgcaaaggg aagggatgct aaggtagagg
     2041 gtgaacgtta cagaaaagca ggctgggaag catatttgag aagatgcggc cagcaaaact
     2101 aaaaaactgt attataagta aatgcatgta tactaaactc acaaattaga gcttcaattt
     2161 aattatatca gttattaccc gagtaacgaa gcatctgtgc ttcattttgt agaacaaaaa
     2221 tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag
     2281 aaatgcaacg cgaaagcgct attttaccaa cgaagaatct gtgcttcatt tttgtaaaac
     2341 aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag
     2401 aacagaaatg caacgcgaga gcgctatttt accaacaaag aatctatact tcttttttgt
     2461 tctacaaaaa tgcatcccga gagcgctatt tttctaacaa agcatcttag attacttttt
     2521 ttctcctttg tgcgctctat aatgcagtct cttgataact ttttgcactg taggtccgtt
     2581 aaggttagaa gaaggctact ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc
     2641 acttcccgcg tttactgatt actagcgaag ctgcgggtgc attttttcaa gataaaggca
     2701 tccccgatta tattctatac cgatgtggat tgcgcatact ttgtgaacag aaagtgatag
     2761 cgttgatgat tcttcattgg tcagaaaatt atgaacggtt tcttctattt tgtctctata
     2821 tactacgtat aggaaatgtt tacattttcg tattgttttc gattcactct atgaatagtt
     2881 cttactacaa tttttttgtc taaagagtaa tactagagat aaacataaaa aatgtagagg
     2941 tcgagtttag atgcaagttc aaggagcgaa aggtggatgg gtaggttata tagggatata
     3001 gcacagagat atatagcaaa gagatacttt tgagcaatgt ttgtggaagc ggtattcgca
     3061 atattttagt agctcgttac agtccggtgc gtttttggtt ttttgaaagt gcgtcatcag
     3121 agcgcttttg gttttcaaaa gcgctctgaa gttcctatac tttctagcta gagaatagga
     3181 acttcccgag cggccgcgtg ttacaagctt gactccagcg taactggact gcaatcaact
     3241 cactggctca ccttcacggg tgggcctttc ttcggtagaa aatcaaagga tcttcttgag
     3301 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3361 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaggtaactg gcttcagcag
     3421 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     3481 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3541 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3601 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3661 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3721 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3781 aggggtaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagca
     3841 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcaga
     3901 aaggcccacc cgaaggtgag ccaggtgatt acatttgggc cctcatcaga ggttttcacc
     3961 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga
     4021 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa
     4081 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactta
     4141 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4201 tgcctgactc cccgtcgtgt agataactac gatgcgggag ggcttaccat ctggccccag
     4261 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4321 gccagccggg agggccgagc gcagaagtga tcctgcaact ttatccgcct ccatccagtc
     4381 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4441 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4501 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4561 tagctccttc ggtcctccga tcgttgccag aagtaagttg gccgcagtgt tatcactcat
     4621 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtgagat gcttttctgt
     4681 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4741 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4801 cattggaaaa cgttcttcgg ggcgtaaact ctcaaggatc ttaccgctgt tgagatccag
     4861 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4921 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4981 gaaatgttga atactcattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa
     5041 tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgccgatc
     5101 aagtcaaaag cctccggtcg gaggcttttg actttctgct atggaggtca ggtatgattt
     5161 aaatggtcag tt
//
