LOCUS       sequence-297725-        4936 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 ORF6 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4936)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4936)
  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-m4046171
FEATURES             Location/Qualifiers
     source          1..4936
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        4..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     CDS             614..631
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             632..649
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(694..712)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1063..1866
                     /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      1949..2928
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3006..3035
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3057..3644
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3656..3683)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3707..3898)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3899..4690,4691..4759))
                     /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     4522..4541
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4760..4862)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4863..4894)
                     /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 atgtttcatc tcgttgactt tcaggttact atagcagaga tattactaat
      481 tattatgagg acttttaaag tttccatttg gaatcttgat tacatcataa acctcataat
      541 taaaaattta tctaagtcac taactgagaa taaatattct caattagatg aagagcaacc
      601 aatggagatt gatctcgtgc cgagaggttc tcaccatcat caccatcatt aaacaggccc
      661 cttttccttt gtcgatatca tgtaattagt tatgtcacgc ttacattcac gccctcctcc
      721 cacatccgct ctaaccgaaa aggaaggagt tagacaacct gaagtctagg tccctattta
      781 ttttttttaa tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt
      841 tttctgtaca aagcggccgc cgaaccacta gtgcactgca gtacacggtt tccttgaaat
      901 ttttttgatt cggtaatctc cgaacagaag gaagaacgaa ggaaggagca cagacttaga
      961 ttggtatata tacgcatatg tagtgttgaa gaaacatgaa attgcccagt attcttaacc
     1021 caactgcaca gaacaaaaac gtgcaggaaa cgaagataaa tcatgtcgaa agctacatat
     1081 aaggaacgtg ctgctactca tcctagtcct gttgctgcca agctatttaa tatcatgcac
     1141 gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta ccaccaagga attactggag
     1201 ttagttgaag cattaggtcc caaaatttgt ttactaaaaa cacatgtgga tatcttgact
     1261 gatttttcca tggagggcac agttaagccg ctaaaggcat tatccgccaa gtacaatttt
     1321 ttactcttcg aggacagaaa atttgctgac attggtaata cagtcaaatt gcagtactct
     1381 gcgggtgtat acagaatagc agaatgggca gacattacga atgcacacgg tgtggtgggc
     1441 ccaggtattg ttagcggttt gaagcaggcg gcagaagaag taacaaagga acctagaggc
     1501 cttttgatgt tagcagaatt gtcatgcaag ggctccctat ctactggaga atatactaag
     1561 ggtactgttg acattgcgaa gagcgacaaa gattttgtta tcggctttat tgctcaaaga
     1621 gacatgggtg gaagagatga aggttacgat tggttgatta tgacacccgg tgtgggttta
     1681 gatgacaagg gagatgcatt gggtcaacag tatagaaccg tggatgatgt ggtttctaca
     1741 ggatctgaca ttattattgt tggaagagga ctatttgcaa agggaaggga tgctaaggta
     1801 gagggtgaac gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa
     1861 aactaaaaaa ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca
     1921 atttaattat atcagttatt acccgagtaa cgaagcatct gtgcttcatt ttgtagaaca
     1981 aaaatgcaac gcgagagcgc taatttttca aacaaagaat ctgagctgca tttttacaga
     2041 acagaaatgc aacgcgaaag cgctatttta ccaacgaaga atctgtgctt catttttgta
     2101 aaacaaaaat gcaacgcgag agcgctaatt tttcaaacaa agaatctgag ctgcattttt
     2161 acagaacaga aatgcaacgc gagagcgcta ttttaccaac aaagaatcta tacttctttt
     2221 ttgttctaca aaaatgcatc ccgagagcgc tattttaaca aagcatctta gattactttt
     2281 tttctccttt gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt
     2341 taaggttaga agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc
     2401 cacttcccgc gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc
     2461 atccccgatt atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata
     2521 gcgttgatga ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat
     2581 atactacgta taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt
     2641 tcttactaca atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag
     2701 gtcgagttta gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat
     2761 agcacagaga tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc
     2821 aatattttag tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcatca
     2881 gagcgctttt ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg
     2941 aacttcccga gcggccgcgt gttacaagct tgactccagc gtaactggac tgcaatcaac
     3001 tcactggctc accttcacgg gtgggccttt cttcggtaga aaatcaaagg atcttcttga
     3061 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3121 gtggtttgtt tgccggatca agagctacca actctttttc cgaggtaact ggcttcagca
     3181 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     3241 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3301 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3361 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     3421 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3481 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3541 caggggtaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3601 atcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcag
     3661 aaaggcccac ccgaaggtga gccaggtgat tacatttggg ccctcatcag aggttttcac
     3721 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     3781 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     3841 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactt
     3901 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3961 ttgcctgact ccccgtcgtg tagataacta cgatgcggga gggcttacca tctggcccca
     4021 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4081 agccagccgg gagggccgag cgcagaagtg atcctgcaac tttatccgcc tccatccagt
     4141 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4201 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4261 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     4321 ttagctcctt cggtcctccg atcgttgcca gaagtaagtt ggccgcagtg ttatcactca
     4381 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtgaga tgcttttctg
     4441 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     4501 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     4561 tcattggaaa acgttcttcg gggcgtaaac tctcaaggat cttaccgctg ttgagatcca
     4621 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     4681 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     4741 ggaaatgttg aatactcatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa
     4801 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat
     4861 caagtcaaaa gcctccggtc ggaggctttt gactttctgc tatggaggtc aggtatgatt
     4921 taaatggtca gttaga
//
