LOCUS       sequence-287559-        4933 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 4933)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4933)
  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-m4046168
FEATURES             Location/Qualifiers
     source          1..4933
                     /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..643
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(688..706)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1057..1860
                     /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      1943..2925
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3003..3032
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3054..3641
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3653..3680)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3704..3895)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3896..4687,4688..4756))
                     /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     4519..4538
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4757..4859)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4860..4891)
                     /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 aggttctgat tataaagatc atgatggtga
      601 ttataaagat catgatattg attataaaga tgatgatgat aaataaacag gccccttttc
      661 ctttgtcgat atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc
      721 cgctctaacc gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt
      781 ttaatagtta tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg
      841 tacaaagcgg ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt
      901 gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta
      961 tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg
     1021 cacagaacaa aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa
     1081 cgtgctgcta ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag
     1141 caaacaaact tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt
     1201 gaagcattag gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt
     1261 tccatggagg gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc
     1321 ttcgaggaca gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt
     1381 gtatacagaa tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt
     1441 attgttagcg gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg
     1501 atgttagcag aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact
     1561 gttgacattg cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg
     1621 ggtggaagag atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac
     1681 aagggagatg cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct
     1741 gacattatta ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt
     1801 gaacgttaca gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa
     1861 aaaactgtat tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa
     1921 ttatatcagt tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg
     1981 caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa
     2041 atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa
     2101 aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa
     2161 cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc
     2221 tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt
     2281 ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa
     2341 ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac
     2401 ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc
     2461 cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg
     2521 ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata
     2581 ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct
     2641 tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc
     2701 gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc
     2761 acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat
     2821 attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag
     2881 cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac
     2941 ttcccgagcg gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca
     3001 ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat
     3061 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3121 gtttgtttgc cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag
     3181 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     3241 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3301 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3361 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3421 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3481 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3541 gggtaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc
     3601 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa
     3661 ggcccacccg aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt
     3721 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
     3781 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
     3841 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc
     3901 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3961 cctgactccc cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg
     4021 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     4081 cagccgggag ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta
     4141 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     4201 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     4261 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     4321 gctccttcgg tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg
     4381 ttatggcagc actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga
     4441 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4501 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4561 ttggaaaacg ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt
     4621 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4681 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4741 aatgttgaat actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata
     4801 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa
     4861 gtcaaaagcc tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa
     4921 atggtcagtt aga
//
