LOCUS       sequence-296382-        6568 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 6568)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6568)
  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-m4046213
FEATURES             Location/Qualifiers
     source          1..6568
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2192..2209
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2210..2233
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(2278..2296)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2647..3450
                     /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      3533..4515
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4593..4622
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4644..5231
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5243..5270)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5294..5485)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5486..6277,6278..6346))
                     /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     6109..6128
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6347..6449)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6450..6481)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        join(6527..6568,1..372)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
ORIGIN
        1 tttactcttc cagattttct cggactccgc gcatcgccgt accacttcaa aacacccaag
       61 cacagcatac taaatttccc ctctttcttc ctctagggtg tcgttaatta cccgtactaa
      121 aggtttggaa aagaaaaaag agaccgcctc gtttcttttt cttcgtcgaa aaaggcaata
      181 aaaattttta tcacgtttct ttttcttgaa aatttttttt tttgattttt ttctctttcg
      241 atgacctccc attgatattt aagttaataa acggtcttca atttctcaag tttcagtttc
      301 atttttcttg ttctattaca acttttttta cttcttgctc attagaaaga aagcatagca
      361 atctaatcta agttttaatt acaaaatggc agtcggcgct tgtgtattat gcaactccca
      421 gacgtcgttg agatgtggtg cttgtattcg tagacctttc ctttgctgta aatgttgcta
      481 tgaccacgtt atctcaactt ctcataagct agttctgagc gtcaatccat acgtgtgtaa
      541 cgcccccggg tgtgatgtta cagacgttac ccaattgtac ttgggaggta tgtcttacta
      601 ttgtaagtcc cacaagccac ctatatcttt tccattatgc gctaacggtc aagtcttcgg
      661 tttgtacaaa aatacctgtg ttggtagtga taatgttacc gactttaacg ccattgctac
      721 ttgtgattgg actaacgcgg gtgattatat cttggctaac acttgtaccg agaggcttaa
      781 gttgttcgcc gctgaaacct taaaggctac ggaagaaaca ttcaaattga gttacggtat
      841 cgctactgtc agagaagttc tatctgatag agaattgcat ctctcctggg aggtgggtaa
      901 gccaagacca ccgttgaacc ggaattacgt ctttactgga tacagggtta ctaagaactc
      961 taaggttcaa attggggaat atacctttga aaagggcgac tacggtgacg ccgtagtcta
     1021 tagaggtacc accacttaca aattgaacgt cggtgactat ttcgttttaa ccagccacac
     1081 agtgatgcca ctgtcagcgc caacattagt tccgcaagag cattacgtta gaattactgg
     1141 tttgtatcca actctaaata tctctgatga attttcctcc aatgtggcta actaccaaaa
     1201 agtaggtatg caaaagtact ctaccttgca aggtccacct ggtactggga agtcgcactt
     1261 cgctataggt ttggcactgt actacccatc tgctagaatt gtttacacgg catgttctca
     1321 tgctgctgta gacgccttgt gtgaaaaggc cttgaaatat ttacccattg ataaatgttc
     1381 aagaattatc ccagcacgag ctcgtgttga atgtttcgac aagtttaagg tcaacagcac
     1441 actagaacaa tacgtcttct gcactgttaa tgctttgcca gaaacgactg ctgatatcgt
     1501 cgtctttgat gaaatctcga tggccactaa ctatgactta tccgtagtta atgctagact
     1561 tcgtgcaaag cactacgtct atattggtga tccagctcag ctgcctgcac caagaacttt
     1621 gttaaccaag ggtaccttgg aaccagaata cttcaactct gtgtgtcgcc tgatgaagac
     1681 aatcggccct gatatgttcc taggtacatg tagacgttgt ccagctgaaa ttgtggacac
     1741 ggtcagtgcc ttggtttacg ataacaaatt aaaggcgcat aaggacaaaa gcgctcaatg
     1801 tttcaagatg ttttacaagg gtgtcattac tcacgatgtt agttcggcca tcaatagacc
     1861 ccaaataggt gttgttagag aatttttgac aaggaaccca gcatggagaa aggctgtttt
     1921 catatcccca tacaattctc aaaacgcggt agcctcaaaa atcttgggtc ttcccactca
     1981 gaccgttgat tcttcccaag gaagtgaata tgactacgtg attttcactc aaaccaccga
     2041 gactgcccac tcttgcaacg tcaatcgttt caacgttgct attacgagag ccaaggttgg
     2101 catcttgtgt atcatgtctg atagagactt gtacgataaa ttacagttta catccttaga
     2161 aataccaaga cgaaacgtcg ctactttgca actcgtgccg agaggttctt ggagccatcc
     2221 gcagtttgaa aaataaacag gccccttttc ctttgtcgat atcatgtaat tagttatgtc
     2281 acgcttacat tcacgccctc ctcccacatc cgctctaacc gaaaaggaag gagttagaca
     2341 acctgaagtc taggtcccta tttatttttt ttaatagtta tgttagtatt aagaacgtta
     2401 tttatatttc aaatttttct tttttttctg tacaaagcgg ccgccgaacc actagtgcac
     2461 tgcagtacac ggtttccttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa
     2521 cgaaggaagg agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca
     2581 tgaaattgcc cagtattctt aacccaactg cacagaacaa aaacgtgcag gaaacgaaga
     2641 taaatcatgt cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct
     2701 gccaagctat ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt
     2761 cgtaccacca aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta
     2821 aaaacacatg tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag
     2881 gcattatccg ccaagtacaa ttttttactc ttcgaggaca gaaaatttgc tgacattggt
     2941 aatacagtca aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt
     3001 acgaatgcac acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa
     3061 gaagtaacaa aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc
     3121 ctatctactg gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt
     3181 gttatcggct ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg
     3241 attatgacac ccggtgtggg tttagatgac aagggagatg cattgggtca acagtataga
     3301 accgtggatg atgtggtttc tacaggatct gacattatta ttgttggaag aggactattt
     3361 gcaaagggaa gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca
     3421 tatttgagaa gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata
     3481 ctaaactcac aaattagagc ttcaatttaa ttatatcagt tattacccga gtaacgaagc
     3541 atctgtgctt cattttgtag aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa
     3601 gaatctgagc tgcattttta cagaacagaa atgcaacgcg aaagcgctat tttaccaacg
     3661 aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa
     3721 acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgagagc gctattttac
     3781 caacaaagaa tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt
     3841 tctaacaaag catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct
     3901 tgataacttt ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt
     3961 ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct
     4021 gcgggtgcat tttttcaaga taaaggcatc cccgattata ttctataccg atgtggattg
     4081 cgcatacttt gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat
     4141 gaacggtttc ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta
     4201 ttgttttcga ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata
     4261 ctagagataa acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag
     4321 gtggatgggt aggttatata gggatatagc acagagatat atagcaaaga gatacttttg
     4381 agcaatgttt gtggaagcgg tattcgcaat attttagtag ctcgttacag tccggtgcgt
     4441 ttttggtttt ttgaaagtgc gtcatcagag cgcttttggt tttcaaaagc gctctgaagt
     4501 tcctatactt tctagctaga gaataggaac ttcccgagcg gccgcgtgtt acaagcttga
     4561 ctccagcgta actggactgc aatcaactca ctggctcacc ttcacgggtg ggcctttctt
     4621 cggtagaaaa tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4681 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4741 ctttttccga ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4801 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4861 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4921 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4981 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5041 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5101 gaacaggaga gcgcacgagg gagcttccag gggtaaacgc ctggtatctt tatagtcctg
     5161 tcgggtttcg ccacctctga cttgagcatc gatttttgtg atgctcgtca ggggggcgga
     5221 gcctatggaa aaacgccagc aacgcagaaa ggcccacccg aaggtgagcc aggtgattac
     5281 atttgggccc tcatcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     5341 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
     5401 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     5461 gggcggtttt ttcctgtttg gtcacttacc aatgcttaat cagtgaggca cctatctcag
     5521 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5581 tgcgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5641 cggctccaga tttatcagca ataaaccagc cagccgggag ggccgagcgc agaagtgatc
     5701 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5761 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5821 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5881 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgccagaa
     5941 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6001 tcatgccatc cgtgagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6061 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6121 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgtaaactct
     6181 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6241 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6301 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatttta gcttccttag
     6361 ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt
     6421 gaaagttgga acctcttacg tgccgatcaa gtcaaaagcc tccggtcgga ggcttttgac
     6481 tttctgctat ggaggtcagg tatgatttaa atggtcagtt agagcggccg ccccacacac
     6541 catagcttca aaatgtttct actccttt
//
