LOCUS       sequence-291440-        6603 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 helicase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6603)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6603)
  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-m4046545
FEATURES             Location/Qualifiers
     source          1..6603
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        25..466
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    25..142
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     437..460
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             2278..2295
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2296..2313
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2358..2376)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2727..3530
                     /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      3613..4595
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4673..4702
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4724..5311
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5323..5350)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5374..5565)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5566..6357,6358..6426))
                     /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     6189..6208
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6427..6529)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6530..6561)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gcggccgctt gctatattga agtacggatt agaagccgcc gagcgggcga cagccctccg
       61 acggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg
      121 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat
      181 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaattaacg aatcaaatta
      241 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag
      301 cgaagcgatg atttttgatc tattaacaga tatataaatg gaaaagctgc ataaccactt
      361 taactaatac tttcaacatt ttcagtttgt attacttctt attcaaatgt cataaaagta
      421 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggagaaaa aactataatg
      481 gttggggctt gtgttctttg caattcacag acttcattaa gatgtggtgc ttgcatacgt
      541 agaccattct tatgttgtaa atgctgttac gaccatgtca tatcaacatc acataaatta
      601 gtcttgtctg ttaatccgta tgtttgcaat gctccaggtt gtgatgtcac agatgtgact
      661 caactttact taggaggtat gagctattat tgtaaatcac ataaaccacc cattagtttt
      721 ccattgtgtg ctaatggaca agtttttggt ttatataaaa atacatgtgt tggtagcgat
      781 aatgttactg actttaatgc aattgcaaca tgtgactgga caaatgctgg tgattacatt
      841 ttagctaaca cctgtactga aagactcaag ctttttgcag cagaaacgct caaagctact
      901 gaggagacat ttaaactgtc ttatggtatt gctactgtac gtgaagtgct gtctgacaga
      961 gaattacatc tttcatggga agttggtaaa cctagaccac cacttaaccg aaattatgtc
     1021 tttactggtt atcgtgtaac taaaaacagt aaagtacaaa taggagagta cacctttgaa
     1081 aaaggtgact atggtgatgc tgttgtttac cgaggtacaa caacttacaa attaaatgtt
     1141 ggtgattatt ttgtgctgac atcacataca gtaatgccat taagtgcacc tacactagtg
     1201 ccacaagagc actatgttag aattactggc ttatacccaa cactcaatat ctcagatgag
     1261 ttttctagca atgttgcaaa ttatcaaaag gttggtatgc aaaagtattc tacactccag
     1321 ggaccacctg gtactggtaa gagtcatttt gctattggcc tagctctcta ctacccttct
     1381 gctcgcatag tgtatacagc ttgctctcat gccgctgttg atgcactatg tgagaaggca
     1441 ttaaaatatt tgcctataga taaatgtagt agaattatac ctgcacgtgc tcgtgtagag
     1501 tgttttgata aattcaaagt gaattcaaca ttagaacagt atgtcttttg tactgtaaat
     1561 gcattgcctg agacgacagc agatatagtt gtctttgatg aaatttcaat ggccacaaat
     1621 tatgatttga gtgttgtcaa tgccagatta cgtgctaagc actatgtgta cattggcgac
     1681 cctgctcaat tacctgcacc acgcacattg ctaactaagg gcacactaga accagaatat
     1741 ttcaattcag tgtgtagact tatgaaaact ataggtccag acatgttcct cggaacttgt
     1801 cggcgttgtc ctgctgaaat tgttgacact gtgagtgctt tggtttatga taataagctt
     1861 aaagcacata aagacaaatc agctcaatgc tttaaaatgt tttataaggg tgttatcacg
     1921 catgatgttt catctgcaat taacaggcca caaataggcg tggtaagaga attccttaca
     1981 cgtaaccctg cttggagaaa agctgtcttt atttcacctt ataattcaca gaatgctgta
     2041 gcctcaaaga ttttgggact accaactcaa actgttgatt catcacaggg ctcagaatat
     2101 gactatgtca tattcactca aaccactgaa acagctcact cttgtaatgt aaacagattt
     2161 aatgttgcta ttaccagagc aaaagtaggc atactttgca taatgtctga tagagacctt
     2221 tatgacaagt tgcaatttac aagtcttgaa attccacgta ggaatgtggc aactttactc
     2281 gtgccgagag gttctcacca tcatcaccat cattaaacag gccccttttc ctttgtcgat
     2341 atcatgtaat tagttatgtc acgcttacat tcacgccctc ctcccacatc cgctctaacc
     2401 gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt ttaatagtta
     2461 tgttagtatt aagaacgtta tttatatttc aaatttttct tttttttctg tacaaagcgg
     2521 ccgccgaacc actagtgcac tgcagtacac ggtttccttg aaattttttt gattcggtaa
     2581 tctccgaaca gaaggaagaa cgaaggaagg agcacagact tagattggta tatatacgca
     2641 tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt aacccaactg cacagaacaa
     2701 aaacgtgcag gaaacgaaga taaatcatgt cgaaagctac atataaggaa cgtgctgcta
     2761 ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag caaacaaact
     2821 tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt gaagcattag
     2881 gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt tccatggagg
     2941 gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc ttcgaggaca
     3001 gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt gtatacagaa
     3061 tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt attgttagcg
     3121 gtttgaagca ggcggcagaa gaagtaacaa aggaacctag aggccttttg atgttagcag
     3181 aattgtcatg caagggctcc ctatctactg gagaatatac taagggtact gttgacattg
     3241 cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg ggtggaagag
     3301 atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac aagggagatg
     3361 cattgggtca acagtataga accgtggatg atgtggtttc tacaggatct gacattatta
     3421 ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt gaacgttaca
     3481 gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa aaaactgtat
     3541 tataagtaaa tgcatgtata ctaaactcac aaattagagc ttcaatttaa ttatatcagt
     3601 tattacccga gtaacgaagc atctgtgctt cattttgtag aacaaaaatg caacgcgaga
     3661 gcgctaattt ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg
     3721 aaagcgctat tttaccaacg aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg
     3781 cgagagcgct aatttttcaa acaaagaatc tgagctgcat ttttacagaa cagaaatgca
     3841 acgcgagagc gctattttac caacaaagaa tctatacttc ttttttgttc tacaaaaatg
     3901 catcccgaga gcgctatttt tctaacaaag catcttagat tacttttttt ctcctttgtg
     3961 cgctctataa tgcagtctct tgataacttt ttgcactgta ggtccgttaa ggttagaaga
     4021 aggctacttt ggtgtctatt ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt
     4081 tactgattac tagcgaagct gcgggtgcat tttttcaaga taaaggcatc cccgattata
     4141 ttctataccg atgtggattg cgcatacttt gtgaacagaa agtgatagcg ttgatgattc
     4201 ttcattggtc agaaaattat gaacggtttc ttctattttg tctctatata ctacgtatag
     4261 gaaatgttta cattttcgta ttgttttcga ttcactctat gaatagttct tactacaatt
     4321 tttttgtcta aagagtaata ctagagataa acataaaaaa tgtagaggtc gagtttagat
     4381 gcaagttcaa ggagcgaaag gtggatgggt aggttatata gggatatagc acagagatat
     4441 atagcaaaga gatacttttg agcaatgttt gtggaagcgg tattcgcaat attttagtag
     4501 ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc gtcatcagag cgcttttggt
     4561 tttcaaaagc gctctgaagt tcctatactt tctagctaga gaataggaac ttcccgagcg
     4621 gccgcgtgtt acaagcttga ctccagcgta actggactgc aatcaactca ctggctcacc
     4681 ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc ttcttgagat cctttttttc
     4741 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4801 cggatcaaga gctaccaact ctttttccga ggtaactggc ttcagcagag cgcagatacc
     4861 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4921 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4981 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5041 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5101 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5161 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag gggtaaacgc
     5221 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcatc gatttttgtg
     5281 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcagaaa ggcccacccg
     5341 aaggtgagcc aggtgattac atttgggccc tcatcagagg ttttcaccgt catcaccgaa
     5401 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc
     5461 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct
     5521 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcacttacc aatgcttaat
     5581 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5641 cgtcgtgtag ataactacga tgcgggaggg cttaccatct ggccccagtg ctgcaatgat
     5701 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccgggag
     5761 ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5821 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5881 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5941 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6001 tcctccgatc gttgccagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6061 actgcataat tctcttactg tcatgccatc cgtgagatgc ttttctgtga ctggtgagta
     6121 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6181 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6241 ttcttcgggg cgtaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6301 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6361 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6421 actcatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag
     6481 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa gtcaaaagcc
     6541 tccggtcgga ggcttttgac tttctgctat ggaggtcagg tatgatttaa atggtcagtt
     6601 aga
//
