LOCUS       sequence-287614-        4885 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 4885)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4885)
  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-m4046591
FEATURES             Location/Qualifiers
     source          1..4885
                     /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             560..577
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             578..595
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(640..658)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1009..1812
                     /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      1895..2877
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      2955..2984
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3006..3593
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3605..3632)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3656..3847)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3848..4639,4640..4708))
                     /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     4471..4490
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4709..4811)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4812..4843)
                     /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 atgattgaac tttcattaat tgacttctat ttgtgctttt tagcctttct
      481 gctattcctt gttttaatta tgcttattat cttttggttc tcacttgaac tgcaagatca
      541 taatgaaact tgtcacgccc tcgtgccgag aggttctcac catcatcacc atcattaaac
      601 aggccccttt tcctttgtcg atatcatgta attagttatg tcacgcttac attcacgccc
      661 tcctcccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc
      721 tatttatttt ttttaatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt
      781 cttttttttc tgtacaaagc ggccgccgaa ccactagtgc actgcagtac acggtttcct
      841 tgaaattttt ttgattcggt aatctccgaa cagaaggaag aacgaaggaa ggagcacaga
      901 cttagattgg tatatatacg catatgtagt gttgaagaaa catgaaattg cccagtattc
      961 ttaacccaac tgcacagaac aaaaacgtgc aggaaacgaa gataaatcat gtcgaaagct
     1021 acatataagg aacgtgctgc tactcatcct agtcctgttg ctgccaagct atttaatatc
     1081 atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg ttcgtaccac caaggaatta
     1141 ctggagttag ttgaagcatt aggtcccaaa atttgtttac taaaaacaca tgtggatatc
     1201 ttgactgatt tttccatgga gggcacagtt aagccgctaa aggcattatc cgccaagtac
     1261 aattttttac tcttcgagga cagaaaattt gctgacattg gtaatacagt caaattgcag
     1321 tactctgcgg gtgtatacag aatagcagaa tgggcagaca ttacgaatgc acacggtgtg
     1381 gtgggcccag gtattgttag cggtttgaag caggcggcag aagaagtaac aaaggaacct
     1441 agaggccttt tgatgttagc agaattgtca tgcaagggct ccctatctac tggagaatat
     1501 actaagggta ctgttgacat tgcgaagagc gacaaagatt ttgttatcgg ctttattgct
     1561 caaagagaca tgggtggaag agatgaaggt tacgattggt tgattatgac acccggtgtg
     1621 ggtttagatg acaagggaga tgcattgggt caacagtata gaaccgtgga tgatgtggtt
     1681 tctacaggat ctgacattat tattgttgga agaggactat ttgcaaaggg aagggatgct
     1741 aaggtagagg gtgaacgtta cagaaaagca ggctgggaag catatttgag aagatgcggc
     1801 cagcaaaact aaaaaactgt attataagta aatgcatgta tactaaactc acaaattaga
     1861 gcttcaattt aattatatca gttattaccc gagtaacgaa gcatctgtgc ttcattttgt
     1921 agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt
     1981 tacagaacag aaatgcaacg cgaaagcgct attttaccaa cgaagaatct gtgcttcatt
     2041 tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc
     2101 atttttacag aacagaaatg caacgcgaga gcgctatttt accaacaaag aatctatact
     2161 tcttttttgt tctacaaaaa tgcatcccga gagcgctatt tttctaacaa agcatcttag
     2221 attacttttt ttctcctttg tgcgctctat aatgcagtct cttgataact ttttgcactg
     2281 taggtccgtt aaggttagaa gaaggctact ttggtgtcta ttttctcttc cataaaaaaa
     2341 gcctgactcc acttcccgcg tttactgatt actagcgaag ctgcgggtgc attttttcaa
     2401 gataaaggca tccccgatta tattctatac cgatgtggat tgcgcatact ttgtgaacag
     2461 aaagtgatag cgttgatgat tcttcattgg tcagaaaatt atgaacggtt tcttctattt
     2521 tgtctctata tactacgtat aggaaatgtt tacattttcg tattgttttc gattcactct
     2581 atgaatagtt cttactacaa tttttttgtc taaagagtaa tactagagat aaacataaaa
     2641 aatgtagagg tcgagtttag atgcaagttc aaggagcgaa aggtggatgg gtaggttata
     2701 tagggatata gcacagagat atatagcaaa gagatacttt tgagcaatgt ttgtggaagc
     2761 ggtattcgca atattttagt agctcgttac agtccggtgc gtttttggtt ttttgaaagt
     2821 gcgtcatcag agcgcttttg gttttcaaaa gcgctctgaa gttcctatac tttctagcta
     2881 gagaatagga acttcccgag cggccgcgtg ttacaagctt gactccagcg taactggact
     2941 gcaatcaact cactggctca ccttcacggg tgggcctttc ttcggtagaa aatcaaagga
     3001 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3061 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaggtaactg
     3121 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     3181 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3241 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3301 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3361 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3421 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     3481 gggagcttcc aggggtaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     3541 gacttgagca tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     3601 gcaacgcaga aaggcccacc cgaaggtgag ccaggtgatt acatttgggc cctcatcaga
     3661 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     3721 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     3781 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     3841 tggtcactta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3901 catccatagt tgcctgactc cccgtcgtgt agataactac gatgcgggag ggcttaccat
     3961 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4021 caataaacca gccagccggg agggccgagc gcagaagtga tcctgcaact ttatccgcct
     4081 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4141 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4201 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4261 aaaaagcggt tagctccttc ggtcctccga tcgttgccag aagtaagttg gccgcagtgt
     4321 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtgagat
     4381 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4441 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4501 aagtgctcat cattggaaaa cgttcttcgg ggcgtaaact ctcaaggatc ttaccgctgt
     4561 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4621 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4681 gggcgacacg gaaatgttga atactcattt tagcttcctt agctcctgaa aatctcgata
     4741 actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta
     4801 cgtgccgatc aagtcaaaag cctccggtcg gaggcttttg actttctgct atggaggtca
     4861 ggtatgattt aaatggtcag ttaga
//
