LOCUS       sequence-313647-        4950 bp    DNA     circular SYN 15-APR-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 4950)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 4950)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pGBW-m4046639
FEATURES             Location/Qualifiers
     source          1..4950
                     /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"
     primer_bind     complement(681..699)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             1050..1853
                     /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      1936..2918
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      2996..3025
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3047..3634
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(3646..3673)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3697..3888)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(3889..4680,4681..4749))
                     /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     4512..4531
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4750..4852)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(4853..4884)
                     /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 attgattaaa caggcccctt ttcctttgtc
      661 gatatcatgt aattagttat gtcacgctta cattcacgcc ctcctcccac atccgctcta
      721 accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt tttttaatag
      781 ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacaaag
      841 cggccgccga accactagtg cactgcagta cacggtttcc ttgaaatttt tttgattcgg
      901 taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg gtatatatac
      961 gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa ctgcacagaa
     1021 caaaaacgtg caggaaacga agataaatca tgtcgaaagc tacatataag gaacgtgctg
     1081 ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa aagcaaacaa
     1141 acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta gttgaagcat
     1201 taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat ttttccatgg
     1261 agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta ctcttcgagg
     1321 acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg ggtgtataca
     1381 gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca ggtattgtta
     1441 gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt ttgatgttag
     1501 cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt actgttgaca
     1561 ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac atgggtggaa
     1621 gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat gacaagggag
     1681 atgcattggg tcaacagtat agaaccgtgg atgatgtggt ttctacagga tctgacatta
     1741 ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt
     1801 acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac taaaaaactg
     1861 tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt taattatatc
     1921 agttattacc cgagtaacga agcatctgtg cttcattttg tagaacaaaa atgcaacgcg
     1981 agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac
     2041 gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa caaaaatgca
     2101 acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca gaacagaaat
     2161 gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg ttctacaaaa
     2221 atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt tttctccttt
     2281 gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt taaggttaga
     2341 agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc cacttcccgc
     2401 gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc atccccgatt
     2461 atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata gcgttgatga
     2521 ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat atactacgta
     2581 taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt tcttactaca
     2641 atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag gtcgagttta
     2701 gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat agcacagaga
     2761 tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc aatattttag
     2821 tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcatca gagcgctttt
     2881 ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg aacttcccga
     2941 gcggccgcgt gttacaagct tgactccagc gtaactggac tgcaatcaac tcactggctc
     3001 accttcacgg gtgggccttt cttcggtaga aaatcaaagg atcttcttga gatccttttt
     3061 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3121 tgccggatca agagctacca actctttttc cgaggtaact ggcttcagca gagcgcagat
     3181 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3241 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3301 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3361 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3421 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3481 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc caggggtaaa
     3541 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc atcgattttt
     3601 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcag aaaggcccac
     3661 ccgaaggtga gccaggtgat tacatttggg ccctcatcag aggttttcac cgtcatcacc
     3721 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     3781 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     3841 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactt accaatgctt
     3901 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3961 ccccgtcgtg tagataacta cgatgcggga gggcttacca tctggcccca gtgctgcaat
     4021 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4081 gagggccgag cgcagaagtg atcctgcaac tttatccgcc tccatccagt ctattaattg
     4141 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4201 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4261 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4321 cggtcctccg atcgttgcca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4381 agcactgcat aattctctta ctgtcatgcc atccgtgaga tgcttttctg tgactggtga
     4441 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4501 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4561 acgttcttcg gggcgtaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4621 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4681 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4741 aatactcatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa atacgcccgg
     4801 tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat caagtcaaaa
     4861 gcctccggtc ggaggctttt gactttctgc tatggaggtc aggtatgatt taaatggtca
     4921 gttagagcgg ccgcttgcta tattgaagta
//
