LOCUS       sequence-307350-        4903 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 4903)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4903)
  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-m4046644
FEATURES             Location/Qualifiers
     source          1..4903
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(577..595)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             946..1749
                     /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      1832..2814
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      2892..2921
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      2943..3530
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3542..3569)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3593..3784)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3785..4576,4577..4645))
                     /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     4408..4427
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4646..4748)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4749..4780)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        join(4826..4903,1..336)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
ORIGIN
        1 ccgtaccact tcaaaacacc caagcacagc atactaaatt tcccctcttt cttcctctag
       61 ggtgtcgtta attacccgta ctaaaggttt ggaaaagaaa aaagagaccg cctcgtttct
      121 ttttcttcgt cgaaaaaggc aataaaaatt tttatcacgt ttctttttct tgaaaatttt
      181 tttttttgat ttttttctct ttcgatgacc tcccattgat atttaagtta ataaacggtc
      241 ttcaatttct caagtttcag tttcattttt cttgttctat tacaactttt tttacttctt
      301 gctcattaga aagaaagcat agcaatctaa tctaagtttt aattacaaaa tgtttcatct
      361 cgttgacttt caggttacta tagcagagat attactaatt attatgagga cttttaaagt
      421 ttccatttgg aatcttgatt acatcataaa cctcataatt aaaaatttat ctaagtcact
      481 aactgagaat aaatattctc aattagatga agagcaacca atggagattg attaaacagg
      541 ccccttttcc tttgtcgata tcatgtaatt agttatgtca cgcttacatt cacgccctcc
      601 tcccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat
      661 ttattttttt taatagttat gttagtatta agaacgttat ttatatttca aatttttctt
      721 ttttttctgt acaaagcggc cgccgaacca ctagtgcact gcagtacacg gtttccttga
      781 aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt
      841 agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta
      901 acccaactgc acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca
      961 tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg
     1021 cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg
     1081 gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg
     1141 actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat
     1201 tttttactct tcgaggacag aaaatttgct gacattggta atacagtcaa attgcagtac
     1261 tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg
     1321 ggcccaggta ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga
     1381 ggccttttga tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact
     1441 aagggtactg ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa
     1501 agagacatgg gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt
     1561 ttagatgaca agggagatgc attgggtcaa cagtatagaa ccgtggatga tgtggtttct
     1621 acaggatctg acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag
     1681 gtagagggtg aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag
     1741 caaaactaaa aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct
     1801 tcaatttaat tatatcagtt attacccgag taacgaagca tctgtgcttc attttgtaga
     1861 acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac
     1921 agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt
     1981 gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     2041 tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct
     2101 tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt
     2161 actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag
     2221 gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc
     2281 tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat
     2341 aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa
     2401 gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt
     2461 ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg
     2521 aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat
     2581 gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag
     2641 ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt
     2701 attcgcaata ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg
     2761 tcatcagagc gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag
     2821 aataggaact tcccgagcgg ccgcgtgtta caagcttgac tccagcgtaa ctggactgca
     2881 atcaactcac tggctcacct tcacgggtgg gcctttcttc ggtagaaaat caaaggatct
     2941 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3001 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgag gtaactggct
     3061 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     3121 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3181 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3241 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3301 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3361 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3421 agcttccagg ggtaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3481 ttgagcatcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3541 acgcagaaag gcccacccga aggtgagcca ggtgattaca tttgggccct catcagaggt
     3601 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt
     3661 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa
     3721 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg
     3781 tcacttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     3841 ccatagttgc ctgactcccc gtcgtgtaga taactacgat gcgggagggc ttaccatctg
     3901 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     3961 taaaccagcc agccgggagg gccgagcgca gaagtgatcc tgcaacttta tccgcctcca
     4021 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4081 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4141 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4201 aagcggttag ctccttcggt cctccgatcg ttgccagaag taagttggcc gcagtgttat
     4261 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtgagatgct
     4321 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4381 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     4441 tgctcatcat tggaaaacgt tcttcggggc gtaaactctc aaggatctta ccgctgttga
     4501 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     4561 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     4621 cgacacggaa atgttgaata ctcattttag cttccttagc tcctgaaaat ctcgataact
     4681 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt
     4741 gccgatcaag tcaaaagcct ccggtcggag gcttttgact ttctgctatg gaggtcaggt
     4801 atgatttaaa tggtcagtta gagcggccgc cccacacacc atagcttcaa aatgtttcta
     4861 ctcctttttt actcttccag attttctcgg actccgcgca tcg
//
