LOCUS       sequence-297580-        6615 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 6615)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6615)
  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-m4046178
FEATURES             Location/Qualifiers
     source          1..6615
                     /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             2284..2301
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2302..2325
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2370..2388)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2739..3542
                     /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      3625..4607
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4685..4714
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4736..5323
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5335..5362)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5386..5577)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5578..6369,6370..6438))
                     /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     6201..6220
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6439..6541)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6542..6573)
                     /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 gcagtcggcg cttgtgtatt atgcaactcc cagacgtcgt tgagatgtgg tgcttgtatt
      541 cgtagacctt tcctttgctg taaatgttgc tatgaccacg ttatctcaac ttctcataag
      601 ctagttctga gcgtcaatcc atacgtgtgt aacgcccccg ggtgtgatgt tacagacgtt
      661 acccaattgt acttgggagg tatgtcttac tattgtaagt cccacaagcc acctatatct
      721 tttccattat gcgctaacgg tcaagtcttc ggtttgtaca aaaatacctg tgttggtagt
      781 gataatgtta ccgactttaa cgccattgct acttgtgatt ggactaacgc gggtgattat
      841 atcttggcta acacttgtac cgagaggctt aagttgttcg ccgctgaaac cttaaaggct
      901 acggaagaaa cattcaaatt gagttacggt atcgctactg tcagagaagt tctatctgat
      961 agagaattgc atctctcctg ggaggtgggt aagccaagac caccgttgaa ccggaattac
     1021 gtctttactg gatacagggt tactaagaac tctaaggttc aaattgggga atataccttt
     1081 gaaaagggcg actacggtga cgccgtagtc tatagaggta ccaccactta caaattgaac
     1141 gtcggtgact atttcgtttt aaccagccac acagtgatgc cactgtcagc gccaacatta
     1201 gttccgcaag agcattacgt tagaattact ggtttgtatc caactctaaa tatctctgat
     1261 gaattttcct ccaatgtggc taactaccaa aaagtaggta tgcaaaagta ctctaccttg
     1321 caaggtccac ctggtactgg gaagtcgcac ttcgctatag gtttggcact gtactaccca
     1381 tctgctagaa ttgtttacac ggcatgttct catgctgctg tagacgcctt gtgtgaaaag
     1441 gccttgaaat atttacccat tgataaatgt tcaagaatta tcccagcacg agctcgtgtt
     1501 gaatgtttcg acaagtttaa ggtcaacagc acactagaac aatacgtctt ctgcactgtt
     1561 aatgctttgc cagaaacgac tgctgatatc gtcgtctttg atgaaatctc gatggccact
     1621 aactatgact tatccgtagt taatgctaga cttcgtgcaa agcactacgt ctatattggt
     1681 gatccagctc agctgcctgc accaagaact ttgttaacca agggtacctt ggaaccagaa
     1741 tacttcaact ctgtgtgtcg cctgatgaag acaatcggcc ctgatatgtt cctaggtaca
     1801 tgtagacgtt gtccagctga aattgtggac acggtcagtg ccttggttta cgataacaaa
     1861 ttaaaggcgc ataaggacaa aagcgctcaa tgtttcaaga tgttttacaa gggtgtcatt
     1921 actcacgatg ttagttcggc catcaataga ccccaaatag gtgttgttag agaatttttg
     1981 acaaggaacc cagcatggag aaaggctgtt ttcatatccc catacaattc tcaaaacgcg
     2041 gtagcctcaa aaatcttggg tcttcccact cagaccgttg attcttccca aggaagtgaa
     2101 tatgactacg tgattttcac tcaaaccacc gagactgccc actcttgcaa cgtcaatcgt
     2161 ttcaacgttg ctattacgag agccaaggtt ggcatcttgt gtatcatgtc tgatagagac
     2221 ttgtacgata aattacagtt tacatcctta gaaataccaa gacgaaacgt cgctactttg
     2281 caactcgtgc cgagaggttc ttggagccat ccgcagtttg aaaaataaac aggccccttt
     2341 tcctttgtcg atatcatgta attagttatg tcacgcttac attcacgccc tcctcccaca
     2401 tccgctctaa ccgaaaagga aggagttaga caacctgaag tctaggtccc tatttatttt
     2461 ttttaatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc
     2521 tgtacaaagc ggccgccgaa ccactagtgc actgcagtac acggtttcct tgaaattttt
     2581 ttgattcggt aatctccgaa cagaaggaag aacgaaggaa ggagcacaga cttagattgg
     2641 tatatatacg catatgtagt gttgaagaaa catgaaattg cccagtattc ttaacccaac
     2701 tgcacagaac aaaaacgtgc aggaaacgaa gataaatcat gtcgaaagct acatataagg
     2761 aacgtgctgc tactcatcct agtcctgttg ctgccaagct atttaatatc atgcacgaaa
     2821 agcaaacaaa cttgtgtgct tcattggatg ttcgtaccac caaggaatta ctggagttag
     2881 ttgaagcatt aggtcccaaa atttgtttac taaaaacaca tgtggatatc ttgactgatt
     2941 tttccatgga gggcacagtt aagccgctaa aggcattatc cgccaagtac aattttttac
     3001 tcttcgagga cagaaaattt gctgacattg gtaatacagt caaattgcag tactctgcgg
     3061 gtgtatacag aatagcagaa tgggcagaca ttacgaatgc acacggtgtg gtgggcccag
     3121 gtattgttag cggtttgaag caggcggcag aagaagtaac aaaggaacct agaggccttt
     3181 tgatgttagc agaattgtca tgcaagggct ccctatctac tggagaatat actaagggta
     3241 ctgttgacat tgcgaagagc gacaaagatt ttgttatcgg ctttattgct caaagagaca
     3301 tgggtggaag agatgaaggt tacgattggt tgattatgac acccggtgtg ggtttagatg
     3361 acaagggaga tgcattgggt caacagtata gaaccgtgga tgatgtggtt tctacaggat
     3421 ctgacattat tattgttgga agaggactat ttgcaaaggg aagggatgct aaggtagagg
     3481 gtgaacgtta cagaaaagca ggctgggaag catatttgag aagatgcggc cagcaaaact
     3541 aaaaaactgt attataagta aatgcatgta tactaaactc acaaattaga gcttcaattt
     3601 aattatatca gttattaccc gagtaacgaa gcatctgtgc ttcattttgt agaacaaaaa
     3661 tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag
     3721 aaatgcaacg cgaaagcgct attttaccaa cgaagaatct gtgcttcatt tttgtaaaac
     3781 aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag
     3841 aacagaaatg caacgcgaga gcgctatttt accaacaaag aatctatact tcttttttgt
     3901 tctacaaaaa tgcatcccga gagcgctatt tttctaacaa agcatcttag attacttttt
     3961 ttctcctttg tgcgctctat aatgcagtct cttgataact ttttgcactg taggtccgtt
     4021 aaggttagaa gaaggctact ttggtgtcta ttttctcttc cataaaaaaa gcctgactcc
     4081 acttcccgcg tttactgatt actagcgaag ctgcgggtgc attttttcaa gataaaggca
     4141 tccccgatta tattctatac cgatgtggat tgcgcatact ttgtgaacag aaagtgatag
     4201 cgttgatgat tcttcattgg tcagaaaatt atgaacggtt tcttctattt tgtctctata
     4261 tactacgtat aggaaatgtt tacattttcg tattgttttc gattcactct atgaatagtt
     4321 cttactacaa tttttttgtc taaagagtaa tactagagat aaacataaaa aatgtagagg
     4381 tcgagtttag atgcaagttc aaggagcgaa aggtggatgg gtaggttata tagggatata
     4441 gcacagagat atatagcaaa gagatacttt tgagcaatgt ttgtggaagc ggtattcgca
     4501 atattttagt agctcgttac agtccggtgc gtttttggtt ttttgaaagt gcgtcatcag
     4561 agcgcttttg gttttcaaaa gcgctctgaa gttcctatac tttctagcta gagaatagga
     4621 acttcccgag cggccgcgtg ttacaagctt gactccagcg taactggact gcaatcaact
     4681 cactggctca ccttcacggg tgggcctttc ttcggtagaa aatcaaagga tcttcttgag
     4741 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4801 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaggtaactg gcttcagcag
     4861 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     4921 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4981 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5041 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5101 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5161 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5221 aggggtaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagca
     5281 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcaga
     5341 aaggcccacc cgaaggtgag ccaggtgatt acatttgggc cctcatcaga ggttttcacc
     5401 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga
     5461 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa
     5521 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactta
     5581 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5641 tgcctgactc cccgtcgtgt agataactac gatgcgggag ggcttaccat ctggccccag
     5701 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5761 gccagccggg agggccgagc gcagaagtga tcctgcaact ttatccgcct ccatccagtc
     5821 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5881 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5941 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     6001 tagctccttc ggtcctccga tcgttgccag aagtaagttg gccgcagtgt tatcactcat
     6061 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtgagat gcttttctgt
     6121 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6181 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6241 cattggaaaa cgttcttcgg ggcgtaaact ctcaaggatc ttaccgctgt tgagatccag
     6301 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6361 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6421 gaaatgttga atactcattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa
     6481 tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgccgatc
     6541 aagtcaaaag cctccggtcg gaggcttttg actttctgct atggaggtca ggtatgattt
     6601 aaatggtcag ttaga
//
