LOCUS       sequence-318502-        5034 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 ORF6 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5034)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 5034)
  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-m4046349
FEATURES             Location/Qualifiers
     source          1..5034
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..442
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    1..118
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     413..436
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             637..654
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             655..720
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(765..783)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1134..1937
                     /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      2020..3002
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3080..3109
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3131..3718
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3730..3757)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3781..3972)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3973..4764,4765..4833))
                     /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     4596..4615
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4834..4936)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4937..4968)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 cggattagaa gccgccgagc gggcgacagc cctccgacgg aagactctcc tccgtgcgtc
       61 ctcgtcttca ccggtcgcgt tcctgaaacg cagatgtgcc tcgcgccgca ctgctccgaa
      121 caataaagat tctacaatac tagcttttat ggttatgaag aggaaaaatt ggcagtaacc
      181 tggccccaca aaccttcaaa ttaacgaatc aaattaacaa ccataggatg ataatgcgat
      241 tagtttttta gccttatttc tggggtaatt aatcagcgaa gcgatgattt ttgatctatt
      301 aacagatata taaatggaaa agctgcataa ccactttaac taatactttc aacattttca
      361 gtttgtatta cttcttattc aaatgtcata aaagtatcaa caaaaaattg ttaatatacc
      421 tctatacttt aacgtcaagg agaaaaaact ataatgtttc atctcgttga ctttcaggtt
      481 actatagcag agatattact aattattatg aggactttta aagtttccat ttggaatctt
      541 gattacatca taaacctcat aattaaaaat ttatctaagt cactaactga gaataaatat
      601 tctcaattag atgaagagca accaatggag attgatctcg tgccgagagg ttctgattat
      661 aaagatcatg atggtgatta taaagatcat gatattgatt ataaagatga tgatgataaa
      721 taaacaggcc ccttttcctt tgtcgatatc atgtaattag ttatgtcacg cttacattca
      781 cgccctcctc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag
      841 gtccctattt atttttttta atagttatgt tagtattaag aacgttattt atatttcaaa
      901 tttttctttt ttttctgtac aaagcggccg ccgaaccact agtgcactgc agtacacggt
      961 ttccttgaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga aggaaggagc
     1021 acagacttag attggtatat atacgcatat gtagtgttga agaaacatga aattgcccag
     1081 tattcttaac ccaactgcac agaacaaaaa cgtgcaggaa acgaagataa atcatgtcga
     1141 aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc aagctattta
     1201 atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt accaccaagg
     1261 aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa acacatgtgg
     1321 atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca ttatccgcca
     1381 agtacaattt tttactcttc gaggacagaa aatttgctga cattggtaat acagtcaaat
     1441 tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg aatgcacacg
     1501 gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa gtaacaaagg
     1561 aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta tctactggag
     1621 aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt atcggcttta
     1681 ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt atgacacccg
     1741 gtgtgggttt agatgacaag ggagatgcat tgggtcaaca gtatagaacc gtggatgatg
     1801 tggtttctac aggatctgac attattattg ttggaagagg actatttgca aagggaaggg
     1861 atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat ttgagaagat
     1921 gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta aactcacaaa
     1981 ttagagcttc aatttaatta tatcagttat tacccgagta acgaagcatc tgtgcttcat
     2041 tttgtagaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc
     2101 atttttacag aacagaaatg caacgcgaaa gcgctatttt accaacgaag aatctgtgct
     2161 tcatttttgt aaaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga
     2221 gctgcatttt tacagaacag aaatgcaacg cgagagcgct attttaccaa caaagaatct
     2281 atacttcttt tttgttctac aaaaatgcat cccgagagcg ctatttttct aacaaagcat
     2341 cttagattac tttttttctc ctttgtgcgc tctataatgc agtctcttga taactttttg
     2401 cactgtaggt ccgttaaggt tagaagaagg ctactttggt gtctattttc tcttccataa
     2461 aaaaagcctg actccacttc ccgcgtttac tgattactag cgaagctgcg ggtgcatttt
     2521 ttcaagataa aggcatcccc gattatattc tataccgatg tggattgcgc atactttgtg
     2581 aacagaaagt gatagcgttg atgattcttc attggtcaga aaattatgaa cggtttcttc
     2641 tattttgtct ctatatacta cgtataggaa atgtttacat tttcgtattg ttttcgattc
     2701 actctatgaa tagttcttac tacaattttt ttgtctaaag agtaatacta gagataaaca
     2761 taaaaaatgt agaggtcgag tttagatgca agttcaagga gcgaaaggtg gatgggtagg
     2821 ttatataggg atatagcaca gagatatata gcaaagagat acttttgagc aatgtttgtg
     2881 gaagcggtat tcgcaatatt ttagtagctc gttacagtcc ggtgcgtttt tggttttttg
     2941 aaagtgcgtc atcagagcgc ttttggtttt caaaagcgct ctgaagttcc tatactttct
     3001 agctagagaa taggaacttc ccgagcggcc gcgtgttaca agcttgactc cagcgtaact
     3061 ggactgcaat caactcactg gctcaccttc acgggtgggc ctttcttcgg tagaaaatca
     3121 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3181 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaggt
     3241 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     3301 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3361 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3421 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     3481 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     3541 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     3601 cacgagggag cttccagggg taaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     3661 cctctgactt gagcatcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     3721 cgccagcaac gcagaaaggc ccacccgaag gtgagccagg tgattacatt tgggccctca
     3781 tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc
     3841 gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt
     3901 ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc
     3961 ctgtttggtc acttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4021 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatgc gggagggctt
     4081 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4141 atcagcaata aaccagccag ccgggagggc cgagcgcaga agtgatcctg caactttatc
     4201 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4261 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4321 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4381 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gccagaagta agttggccgc
     4441 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4501 gagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4561 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4621 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcgt aaactctcaa ggatcttacc
     4681 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4741 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4801 aataagggcg acacggaaat gttgaatact cattttagct tccttagctc ctgaaaatct
     4861 cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc
     4921 tcttacgtgc cgatcaagtc aaaagcctcc ggtcggaggc ttttgacttt ctgctatgga
     4981 ggtcaggtat gatttaaatg gtcagttaga gcggccgctt gctatattga agta
//
