LOCUS       sequence-291580-        6610 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 6610)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6610)
  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-m4046250
FEATURES             Location/Qualifiers
     source          1..6610
                     /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..2320
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2365..2383)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2734..3537
                     /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      3620..4602
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4680..4709
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4731..5318
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5330..5357)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5381..5572)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5573..6364,6365..6433))
                     /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     6196..6215
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6434..6536)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6537..6568)
                     /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 ttctgattat aaagatcatg atggtgatta
     2281 taaagatcat gatattgatt ataaagatga tgatgataaa taaacaggcc ccttttcctt
     2341 tgtcgatatc atgtaattag ttatgtcacg cttacattca cgccctcctc ccacatccgc
     2401 tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttta
     2461 atagttatgt tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac
     2521 aaagcggccg ccgaaccact agtgcactgc agtacacggt ttccttgaaa tttttttgat
     2581 tcggtaatct ccgaacagaa ggaagaacga aggaaggagc acagacttag attggtatat
     2641 atacgcatat gtagtgttga agaaacatga aattgcccag tattcttaac ccaactgcac
     2701 agaacaaaaa cgtgcaggaa acgaagataa atcatgtcga aagctacata taaggaacgt
     2761 gctgctactc atcctagtcc tgttgctgcc aagctattta atatcatgca cgaaaagcaa
     2821 acaaacttgt gtgcttcatt ggatgttcgt accaccaagg aattactgga gttagttgaa
     2881 gcattaggtc ccaaaatttg tttactaaaa acacatgtgg atatcttgac tgatttttcc
     2941 atggagggca cagttaagcc gctaaaggca ttatccgcca agtacaattt tttactcttc
     3001 gaggacagaa aatttgctga cattggtaat acagtcaaat tgcagtactc tgcgggtgta
     3061 tacagaatag cagaatgggc agacattacg aatgcacacg gtgtggtggg cccaggtatt
     3121 gttagcggtt tgaagcaggc ggcagaagaa gtaacaaagg aacctagagg ccttttgatg
     3181 ttagcagaat tgtcatgcaa gggctcccta tctactggag aatatactaa gggtactgtt
     3241 gacattgcga agagcgacaa agattttgtt atcggcttta ttgctcaaag agacatgggt
     3301 ggaagagatg aaggttacga ttggttgatt atgacacccg gtgtgggttt agatgacaag
     3361 ggagatgcat tgggtcaaca gtatagaacc gtggatgatg tggtttctac aggatctgac
     3421 attattattg ttggaagagg actatttgca aagggaaggg atgctaaggt agagggtgaa
     3481 cgttacagaa aagcaggctg ggaagcatat ttgagaagat gcggccagca aaactaaaaa
     3541 actgtattat aagtaaatgc atgtatacta aactcacaaa ttagagcttc aatttaatta
     3601 tatcagttat tacccgagta acgaagcatc tgtgcttcat tttgtagaac aaaaatgcaa
     3661 cgcgagagcg ctaatttttc aaacaaagaa tctgagctgc atttttacag aacagaaatg
     3721 caacgcgaaa gcgctatttt accaacgaag aatctgtgct tcatttttgt aaaacaaaaa
     3781 tgcaacgcga gagcgctaat ttttcaaaca aagaatctga gctgcatttt tacagaacag
     3841 aaatgcaacg cgagagcgct attttaccaa caaagaatct atacttcttt tttgttctac
     3901 aaaaatgcat cccgagagcg ctatttttct aacaaagcat cttagattac tttttttctc
     3961 ctttgtgcgc tctataatgc agtctcttga taactttttg cactgtaggt ccgttaaggt
     4021 tagaagaagg ctactttggt gtctattttc tcttccataa aaaaagcctg actccacttc
     4081 ccgcgtttac tgattactag cgaagctgcg ggtgcatttt ttcaagataa aggcatcccc
     4141 gattatattc tataccgatg tggattgcgc atactttgtg aacagaaagt gatagcgttg
     4201 atgattcttc attggtcaga aaattatgaa cggtttcttc tattttgtct ctatatacta
     4261 cgtataggaa atgtttacat tttcgtattg ttttcgattc actctatgaa tagttcttac
     4321 tacaattttt ttgtctaaag agtaatacta gagataaaca taaaaaatgt agaggtcgag
     4381 tttagatgca agttcaagga gcgaaaggtg gatgggtagg ttatataggg atatagcaca
     4441 gagatatata gcaaagagat acttttgagc aatgtttgtg gaagcggtat tcgcaatatt
     4501 ttagtagctc gttacagtcc ggtgcgtttt tggttttttg aaagtgcgtc atcagagcgc
     4561 ttttggtttt caaaagcgct ctgaagttcc tatactttct agctagagaa taggaacttc
     4621 ccgagcggcc gcgtgttaca agcttgactc cagcgtaact ggactgcaat caactcactg
     4681 gctcaccttc acgggtgggc ctttcttcgg tagaaaatca aaggatcttc ttgagatcct
     4741 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4801 tgtttgccgg atcaagagct accaactctt tttccgaggt aactggcttc agcagagcgc
     4861 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     4921 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4981 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5041 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5101 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5161 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5221 taaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcatcgat
     5281 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcagaaaggc
     5341 ccacccgaag gtgagccagg tgattacatt tgggccctca tcagaggttt tcaccgtcat
     5401 caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac
     5461 agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct
     5521 ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc acttaccaat
     5581 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5641 gactccccgt cgtgtagata actacgatgc gggagggctt accatctggc cccagtgctg
     5701 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5761 ccgggagggc cgagcgcaga agtgatcctg caactttatc cgcctccatc cagtctatta
     5821 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5881 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5941 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6001 ccttcggtcc tccgatcgtt gccagaagta agttggccgc agtgttatca ctcatggtta
     6061 tggcagcact gcataattct cttactgtca tgccatccgt gagatgcttt tctgtgactg
     6121 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6181 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6241 gaaaacgttc ttcggggcgt aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6301 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6361 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6421 gttgaatact cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc
     6481 ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaagtc
     6541 aaaagcctcc ggtcggaggc ttttgacttt ctgctatgga ggtcaggtat gatttaaatg
     6601 gtcagttaga
//
