LOCUS       sequence-291464-        4939 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 4939)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4939)
  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..4939
                     /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             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..2931
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      3009..3038
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3060..3647
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3659..3686)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3710..3901)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3902..4693,4694..4762))
                     /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     4525..4544
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4763..4865)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4866..4897)
                     /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 tatttttcta acaaagcatc ttagattact
     2281 ttttttctcc tttgtgcgct ctataatgca gtctcttgat aactttttgc actgtaggtc
     2341 cgttaaggtt agaagaaggc tactttggtg tctattttct cttccataaa aaaagcctga
     2401 ctccacttcc cgcgtttact gattactagc gaagctgcgg gtgcattttt tcaagataaa
     2461 ggcatccccg attatattct ataccgatgt ggattgcgca tactttgtga acagaaagtg
     2521 atagcgttga tgattcttca ttggtcagaa aattatgaac ggtttcttct attttgtctc
     2581 tatatactac gtataggaaa tgtttacatt ttcgtattgt tttcgattca ctctatgaat
     2641 agttcttact acaatttttt tgtctaaaga gtaatactag agataaacat aaaaaatgta
     2701 gaggtcgagt ttagatgcaa gttcaaggag cgaaaggtgg atgggtaggt tatataggga
     2761 tatagcacag agatatatag caaagagata cttttgagca atgtttgtgg aagcggtatt
     2821 cgcaatattt tagtagctcg ttacagtccg gtgcgttttt ggttttttga aagtgcgtca
     2881 tcagagcgct tttggttttc aaaagcgctc tgaagttcct atactttcta gctagagaat
     2941 aggaacttcc cgagcggccg cgtgttacaa gcttgactcc agcgtaactg gactgcaatc
     3001 aactcactgg ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa aggatcttct
     3061 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3121 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaggta actggcttca
     3181 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     3241 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3301 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3361 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3421 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3481 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3541 ttccaggggt aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3601 agcatcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3661 cagaaaggcc cacccgaagg tgagccaggt gattacattt gggccctcat cagaggtttt
     3721 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
     3781 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     3841 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     3901 cttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     3961 tagttgcctg actccccgtc gtgtagataa ctacgatgcg ggagggctta ccatctggcc
     4021 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4081 accagccagc cgggagggcc gagcgcagaa gtgatcctgc aactttatcc gcctccatcc
     4141 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     4201 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     4261 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     4321 cggttagctc cttcggtcct ccgatcgttg ccagaagtaa gttggccgca gtgttatcac
     4381 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgtg agatgctttt
     4441 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     4501 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     4561 tcatcattgg aaaacgttct tcggggcgta aactctcaag gatcttaccg ctgttgagat
     4621 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     4681 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     4741 cacggaaatg ttgaatactc attttagctt ccttagctcc tgaaaatctc gataactcaa
     4801 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
     4861 gatcaagtca aaagcctccg gtcggaggct tttgactttc tgctatggag gtcaggtatg
     4921 atttaaatgg tcagttaga
//
