LOCUS       sequence-297242-        7636 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 7636)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 7636)
  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-m4046280
FEATURES             Location/Qualifiers
     source          1..7636
                     /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             2231..2248
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(3391..3409)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             3760..4563
                     /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      4646..5628
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      5706..5735
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      5757..6344
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(6356..6383)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(6407..6598)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6599..7390,7391..7459))
                     /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     7222..7241
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7460..7562)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(7563..7594)
                     /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 atggttgggg cttgtgttct ttgcaattca cagacttcat taagatgtgg
      481 tgcttgcata cgtagaccat tcttatgttg taaatgctgt tacgaccatg tcatatcaac
      541 atcacataaa ttagtcttgt ctgttaatcc gtatgtttgc aatgctccag gttgtgatgt
      601 cacagatgtg actcaacttt acttaggagg tatgagctat tattgtaaat cacataaacc
      661 acccattagt tttccattgt gtgctaatgg acaagttttt ggtttatata aaaatacatg
      721 tgttggtagc gataatgtta ctgactttaa tgcaattgca acatgtgact ggacaaatgc
      781 tggtgattac attttagcta acacctgtac tgaaagactc aagctttttg cagcagaaac
      841 gctcaaagct actgaggaga catttaaact gtcttatggt attgctactg tacgtgaagt
      901 gctgtctgac agagaattac atctttcatg ggaagttggt aaacctagac caccacttaa
      961 ccgaaattat gtctttactg gttatcgtgt aactaaaaac agtaaagtac aaataggaga
     1021 gtacaccttt gaaaaaggtg actatggtga tgctgttgtt taccgaggta caacaactta
     1081 caaattaaat gttggtgatt attttgtgct gacatcacat acagtaatgc cattaagtgc
     1141 acctacacta gtgccacaag agcactatgt tagaattact ggcttatacc caacactcaa
     1201 tatctcagat gagttttcta gcaatgttgc aaattatcaa aaggttggta tgcaaaagta
     1261 ttctacactc cagggaccac ctggtactgg taagagtcat tttgctattg gcctagctct
     1321 ctactaccct tctgctcgca tagtgtatac agcttgctct catgccgctg ttgatgcact
     1381 atgtgagaag gcattaaaat atttgcctat agataaatgt agtagaatta tacctgcacg
     1441 tgctcgtgta gagtgttttg ataaattcaa agtgaattca acattagaac agtatgtctt
     1501 ttgtactgta aatgcattgc ctgagacgac agcagatata gttgtctttg atgaaatttc
     1561 aatggccaca aattatgatt tgagtgttgt caatgccaga ttacgtgcta agcactatgt
     1621 gtacattggc gaccctgctc aattacctgc accacgcaca ttgctaacta agggcacact
     1681 agaaccagaa tatttcaatt cagtgtgtag acttatgaaa actataggtc cagacatgtt
     1741 cctcggaact tgtcggcgtt gtcctgctga aattgttgac actgtgagtg ctttggttta
     1801 tgataataag cttaaagcac ataaagacaa atcagctcaa tgctttaaaa tgttttataa
     1861 gggtgttatc acgcatgatg tttcatctgc aattaacagg ccacaaatag gcgtggtaag
     1921 agaattcctt acacgtaacc ctgcttggag aaaagctgtc tttatttcac cttataattc
     1981 acagaatgct gtagcctcaa agattttggg actaccaact caaactgttg attcatcaca
     2041 gggctcagaa tatgactatg tcatattcac tcaaaccact gaaacagctc actcttgtaa
     2101 tgtaaacaga tttaatgttg ctattaccag agcaaaagta ggcatacttt gcataatgtc
     2161 tgatagagac ctttatgaca agttgcaatt tacaagtctt gaaattccac gtaggaatgt
     2221 ggcaacttta ctcgtgccga gaggttctaa aatcgaggaa ggtaagctag tcatttggat
     2281 aaatggagat aagggttaca acggtttggc agaagtaggc aagaaatttg aaaaggacac
     2341 aggtatcaag gtgacggttg aacatccaga caagttagaa gaaaaattcc ctcaagttgc
     2401 tgctactggt gatggtcccg acattatttt ctgggcccac gatcgttttg gtggatatgc
     2461 tcagtcaggt ttgctggctg agatcacccc agacaaggcc ttccaagata aattgtaccc
     2521 attcacttgg gatgctgtca gatacaacgg taagcttata gcatatccaa ttgctgttga
     2581 agccttatcg ttgatctaca ataaggactt gttacctaac ccaccaaaga catgggaaga
     2641 aattccagct ttggataaag agctaaaggc caagggaaaa agcgctttga tgtttaactt
     2701 acaagaacct tatttcacct ggccattaat cgctgctgac ggtggttacg catttaaata
     2761 cgaaaacggt aagtatgaca ttaaggatgt cggcgttgat aatgctggtg caaaggctgg
     2821 tttgaccttc ttggttgatt taataaagaa taaacacatg aacgctgaca ctgactactc
     2881 tatcgccgaa gccgctttca acaagggtga aactgctatg accataaatg gtccatgggc
     2941 ctggagtaac attgatactt ctaaagtaaa ctatggtgtc actgttttgc ctacattcaa
     3001 aggccaacca tccaagccct ttgtcggtgt tctatccgct ggcatcaatg ctgcttctcc
     3061 aaacaaggag ctagctaaag aatttttgga aaattacttg ttaactgacg aaggtttgga
     3121 ggccgttaac aaggacaagc cattgggtgc tgtcgctttg aaatcttacg aagaagaatt
     3181 ggctaaggat cccagaattg ccgctaccat ggagaacgcc caaaagggtg aaattatgcc
     3241 aaacatccca caaatgtcag ccttctggta cgcagttagg actgccgtga ttaatgctgc
     3301 ttccggtaga caaaccgtcg acgaagcatt aaaagatgct caaacgtaaa caggcccctt
     3361 ttcctttgtc gatatcatgt aattagttat gtcacgctta cattcacgcc ctcctcccac
     3421 atccgctcta accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt
     3481 tttttaatag ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt
     3541 ctgtacaaag cggccgccga accactagtg cactgcagta cacggtttcc ttgaaatttt
     3601 tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag acttagattg
     3661 gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt cttaacccaa
     3721 ctgcacagaa caaaaacgtg caggaaacga agataaatca tgtcgaaagc tacatataag
     3781 gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat catgcacgaa
     3841 aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt actggagtta
     3901 gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat cttgactgat
     3961 ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta caatttttta
     4021 ctcttcgagg acagaaaatt tgctgacatt ggtaatacag tcaaattgca gtactctgcg
     4081 ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt ggtgggccca
     4141 ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc tagaggcctt
     4201 ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata tactaagggt
     4261 actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc tcaaagagac
     4321 atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt gggtttagat
     4381 gacaagggag atgcattggg tcaacagtat agaaccgtgg atgatgtggt ttctacagga
     4441 tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc taaggtagag
     4501 ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg ccagcaaaac
     4561 taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag agcttcaatt
     4621 taattatatc agttattacc cgagtaacga agcatctgtg cttcattttg tagaacaaaa
     4681 atgcaacgcg agagcgctaa tttttcaaac aaagaatctg agctgcattt ttacagaaca
     4741 gaaatgcaac gcgaaagcgc tattttacca acgaagaatc tgtgcttcat ttttgtaaaa
     4801 caaaaatgca acgcgagagc gctaattttt caaacaaaga atctgagctg catttttaca
     4861 gaacagaaat gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg
     4921 ttctacaaaa atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt
     4981 tttctccttt gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt
     5041 taaggttaga agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc
     5101 cacttcccgc gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc
     5161 atccccgatt atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata
     5221 gcgttgatga ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat
     5281 atactacgta taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt
     5341 tcttactaca atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag
     5401 gtcgagttta gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat
     5461 agcacagaga tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc
     5521 aatattttag tagctcgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcatca
     5581 gagcgctttt ggttttcaaa agcgctctga agttcctata ctttctagct agagaatagg
     5641 aacttcccga gcggccgcgt gttacaagct tgactccagc gtaactggac tgcaatcaac
     5701 tcactggctc accttcacgg gtgggccttt cttcggtaga aaatcaaagg atcttcttga
     5761 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5821 gtggtttgtt tgccggatca agagctacca actctttttc cgaggtaact ggcttcagca
     5881 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     5941 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6001 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6061 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6121 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6181 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6241 caggggtaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6301 atcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcag
     6361 aaaggcccac ccgaaggtga gccaggtgat tacatttggg ccctcatcag aggttttcac
     6421 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     6481 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     6541 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactt
     6601 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     6661 ttgcctgact ccccgtcgtg tagataacta cgatgcggga gggcttacca tctggcccca
     6721 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     6781 agccagccgg gagggccgag cgcagaagtg atcctgcaac tttatccgcc tccatccagt
     6841 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     6901 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6961 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7021 ttagctcctt cggtcctccg atcgttgcca gaagtaagtt ggccgcagtg ttatcactca
     7081 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtgaga tgcttttctg
     7141 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7201 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7261 tcattggaaa acgttcttcg gggcgtaaac tctcaaggat cttaccgctg ttgagatcca
     7321 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7381 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7441 ggaaatgttg aatactcatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa
     7501 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat
     7561 caagtcaaaa gcctccggtc ggaggctttt gactttctgc tatggaggtc aggtatgatt
     7621 taaatggtca gttaga
//
