LOCUS       sequence-298031-        6562 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 6562)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6562)
  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-m4046232
FEATURES             Location/Qualifiers
     source          1..6562
                     /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             2237..2254
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2255..2272
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2317..2335)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2686..3489
                     /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      3572..4554
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4632..4661
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4683..5270
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5282..5309)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5333..5524)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5525..6316,6317..6385))
                     /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     6148..6167
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6386..6488)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6489..6520)
                     /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 atggcagtcg gcgcttgtgt attatgcaac tcccagacgt cgttgagatg
      481 tggtgcttgt attcgtagac ctttcctttg ctgtaaatgt tgctatgacc acgttatctc
      541 aacttctcat aagctagttc tgagcgtcaa tccatacgtg tgtaacgccc ccgggtgtga
      601 tgttacagac gttacccaat tgtacttggg aggtatgtct tactattgta agtcccacaa
      661 gccacctata tcttttccat tatgcgctaa cggtcaagtc ttcggtttgt acaaaaatac
      721 ctgtgttggt agtgataatg ttaccgactt taacgccatt gctacttgtg attggactaa
      781 cgcgggtgat tatatcttgg ctaacacttg taccgagagg cttaagttgt tcgccgctga
      841 aaccttaaag gctacggaag aaacattcaa attgagttac ggtatcgcta ctgtcagaga
      901 agttctatct gatagagaat tgcatctctc ctgggaggtg ggtaagccaa gaccaccgtt
      961 gaaccggaat tacgtcttta ctggatacag ggttactaag aactctaagg ttcaaattgg
     1021 ggaatatacc tttgaaaagg gcgactacgg tgacgccgta gtctatagag gtaccaccac
     1081 ttacaaattg aacgtcggtg actatttcgt tttaaccagc cacacagtga tgccactgtc
     1141 agcgccaaca ttagttccgc aagagcatta cgttagaatt actggtttgt atccaactct
     1201 aaatatctct gatgaatttt cctccaatgt ggctaactac caaaaagtag gtatgcaaaa
     1261 gtactctacc ttgcaaggtc cacctggtac tgggaagtcg cacttcgcta taggtttggc
     1321 actgtactac ccatctgcta gaattgttta cacggcatgt tctcatgctg ctgtagacgc
     1381 cttgtgtgaa aaggccttga aatatttacc cattgataaa tgttcaagaa ttatcccagc
     1441 acgagctcgt gttgaatgtt tcgacaagtt taaggtcaac agcacactag aacaatacgt
     1501 cttctgcact gttaatgctt tgccagaaac gactgctgat atcgtcgtct ttgatgaaat
     1561 ctcgatggcc actaactatg acttatccgt agttaatgct agacttcgtg caaagcacta
     1621 cgtctatatt ggtgatccag ctcagctgcc tgcaccaaga actttgttaa ccaagggtac
     1681 cttggaacca gaatacttca actctgtgtg tcgcctgatg aagacaatcg gccctgatat
     1741 gttcctaggt acatgtagac gttgtccagc tgaaattgtg gacacggtca gtgccttggt
     1801 ttacgataac aaattaaagg cgcataagga caaaagcgct caatgtttca agatgtttta
     1861 caagggtgtc attactcacg atgttagttc ggccatcaat agaccccaaa taggtgttgt
     1921 tagagaattt ttgacaagga acccagcatg gagaaaggct gttttcatat ccccatacaa
     1981 ttctcaaaac gcggtagcct caaaaatctt gggtcttccc actcagaccg ttgattcttc
     2041 ccaaggaagt gaatatgact acgtgatttt cactcaaacc accgagactg cccactcttg
     2101 caacgtcaat cgtttcaacg ttgctattac gagagccaag gttggcatct tgtgtatcat
     2161 gtctgataga gacttgtacg ataaattaca gtttacatcc ttagaaatac caagacgaaa
     2221 cgtcgctact ttgcaactcg tgccgagagg ttctcaccat catcaccatc attaaacagg
     2281 ccccttttcc tttgtcgata tcatgtaatt agttatgtca cgcttacatt cacgccctcc
     2341 tcccacatcc gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat
     2401 ttattttttt taatagttat gttagtatta agaacgttat ttatatttca aatttttctt
     2461 ttttttctgt acaaagcggc cgccgaacca ctagtgcact gcagtacacg gtttccttga
     2521 aatttttttg attcggtaat ctccgaacag aaggaagaac gaaggaagga gcacagactt
     2581 agattggtat atatacgcat atgtagtgtt gaagaaacat gaaattgccc agtattctta
     2641 acccaactgc acagaacaaa aacgtgcagg aaacgaagat aaatcatgtc gaaagctaca
     2701 tataaggaac gtgctgctac tcatcctagt cctgttgctg ccaagctatt taatatcatg
     2761 cacgaaaagc aaacaaactt gtgtgcttca ttggatgttc gtaccaccaa ggaattactg
     2821 gagttagttg aagcattagg tcccaaaatt tgtttactaa aaacacatgt ggatatcttg
     2881 actgattttt ccatggaggg cacagttaag ccgctaaagg cattatccgc caagtacaat
     2941 tttttactct tcgaggacag aaaatttgct gacattggta atacagtcaa attgcagtac
     3001 tctgcgggtg tatacagaat agcagaatgg gcagacatta cgaatgcaca cggtgtggtg
     3061 ggcccaggta ttgttagcgg tttgaagcag gcggcagaag aagtaacaaa ggaacctaga
     3121 ggccttttga tgttagcaga attgtcatgc aagggctccc tatctactgg agaatatact
     3181 aagggtactg ttgacattgc gaagagcgac aaagattttg ttatcggctt tattgctcaa
     3241 agagacatgg gtggaagaga tgaaggttac gattggttga ttatgacacc cggtgtgggt
     3301 ttagatgaca agggagatgc attgggtcaa cagtatagaa ccgtggatga tgtggtttct
     3361 acaggatctg acattattat tgttggaaga ggactatttg caaagggaag ggatgctaag
     3421 gtagagggtg aacgttacag aaaagcaggc tgggaagcat atttgagaag atgcggccag
     3481 caaaactaaa aaactgtatt ataagtaaat gcatgtatac taaactcaca aattagagct
     3541 tcaatttaat tatatcagtt attacccgag taacgaagca tctgtgcttc attttgtaga
     3601 acaaaaatgc aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac
     3661 agaacagaaa tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt
     3721 gtaaaacaaa aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt
     3781 tttacagaac agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct
     3841 tttttgttct acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt
     3901 actttttttc tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag
     3961 gtccgttaag gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc
     4021 tgactccact tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat
     4081 aaaggcatcc ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa
     4141 gtgatagcgt tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt
     4201 ctctatatac tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg
     4261 aatagttctt actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat
     4321 gtagaggtcg agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag
     4381 ggatatagca cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt
     4441 attcgcaata ttttagtagc tcgttacagt ccggtgcgtt tktggttttt tgaaagtgcg
     4501 tcatcagagc gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag
     4561 aataggaact tcccgagcgg ccgcgtgtta caagcttgac tccagcgtaa ctggactgca
     4621 atcaactcac tggctcacct tcacgggtgg gcctttcttc ggtagaaaat caaaggatct
     4681 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4741 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgag gtaactggct
     4801 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     4861 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4921 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4981 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5041 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5101 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5161 agcttccagg ggtaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5221 ttgagcatcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5281 acgcagaaag gcccacccga aggtgagcca ggtgattaca tttgggccct catcagaggt
     5341 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt
     5401 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa
     5461 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg
     5521 tcacttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5581 ccatagttgc ctgactcccc gtcgtgtaga taactacgat gcgggagggc ttaccatctg
     5641 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5701 taaaccagcc agccgggagg gccgagcgca gaagtgatcc tgcaacttta tccgcctcca
     5761 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5821 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5881 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5941 aagcggttag ctccttcggt cctccgatcg ttgccagaag taagttggcc gcagtgttat
     6001 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtgagatgct
     6061 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6121 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6181 tgctcatcat tggaaaacgt tcttcggggc gtaaactctc aaggatctta ccgctgttga
     6241 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6301 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6361 cgacacggaa atgttgaata ctcattttag cttccttagc tcctgaaaat ctcgataact
     6421 caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa cctcttacgt
     6481 gccgatcaag tcaaaagcct ccggtcggag gcttttgact ttctgctatg gaggtcaggt
     6541 atgatttaaa tggtcagtta ga
//
