LOCUS       sequence-297630-        6520 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 6520)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6520)
  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-m4046638
FEATURES             Location/Qualifiers
     source          1..6520
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..417
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(2275..2293)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2644..3447
                     /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      3530..4512
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4590..4619
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4641..5228
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5240..5267)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5291..5482)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5483..6274,6275..6343))
                     /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     6106..6125
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6344..6446)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6447..6478)
                     /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 taaacaggcc ccttttcctt tgtcgatatc atgtaattag ttatgtcacg
     2281 cttacattca cgccctcctc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc
     2341 tgaagtctag gtccctattt atttttttta atagttatgt tagtattaag aacgttattt
     2401 atatttcaaa tttttctttt ttttctgtac aaagcggccg ccgaaccact agtgcactgc
     2461 agtacacggt ttccttgaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga
     2521 aggaaggagc acagacttag attggtatat atacgcatat gtagtgttga agaaacatga
     2581 aattgcccag tattcttaac ccaactgcac agaacaaaaa cgtgcaggaa acgaagataa
     2641 atcatgtcga aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc
     2701 aagctattta atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt
     2761 accaccaagg aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa
     2821 acacatgtgg atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca
     2881 ttatccgcca agtacaattt tttactcttc gaggacagaa aatttgctga cattggtaat
     2941 acagtcaaat tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg
     3001 aatgcacacg gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa
     3061 gtaacaaagg aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta
     3121 tctactggag aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt
     3181 atcggcttta ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt
     3241 atgacacccg gtgtgggttt agatgacaag ggagatgcat tgggtcaaca gtatagaacc
     3301 gtggatgatg tggtttctac aggatctgac attattattg ttggaagagg actatttgca
     3361 aagggaaggg atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat
     3421 ttgagaagat gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta
     3481 aactcacaaa ttagagcttc aatttaatta tatcagttat tacccgagta acgaagcatc
     3541 tgtgcttcat tttgtagaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa
     3601 tctgagctgc atttttacag aacagaaatg caacgcgaaa gcgctatttt accaacgaag
     3661 aatctgtgct tcatttttgt aaaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca
     3721 aagaatctga gctgcatttt tacagaacag aaatgcaacg cgagagcgct attttaccaa
     3781 caaagaatct atacttcttt tttgttctac aaaaatgcat cccgagagcg ctatttttct
     3841 aacaaagcat cttagattac tttttttctc ctttgtgcgc tctataatgc agtctcttga
     3901 taactttttg cactgtaggt ccgttaaggt tagaagaagg ctactttggt gtctattttc
     3961 tcttccataa aaaaagcctg actccacttc ccgcgtttac tgattactag cgaagctgcg
     4021 ggtgcatttt ttcaagataa aggcatcccc gattatattc tataccgatg tggattgcgc
     4081 atactttgtg aacagaaagt gatagcgttg atgattcttc attggtcaga aaattatgaa
     4141 cggtttcttc tattttgtct ctatatacta cgtataggaa atgtttacat tttcgtattg
     4201 ttttcgattc actctatgaa tagttcttac tacaattttt ttgtctaaag agtaatacta
     4261 gagataaaca taaaaaatgt agaggtcgag tttagatgca agttcaagga gcgaaaggtg
     4321 gatgggtagg ttatataggg atatagcaca gagatatata gcaaagagat acttttgagc
     4381 aatgtttgtg gaagcggtat tcgcaatatt ttagtagctc gttacagtcc ggtgcgtttt
     4441 tggttttttg aaagtgcgtc atcagagcgc ttttggtttt caaaagcgct ctgaagttcc
     4501 tatactttct agctagagaa taggaacttc ccgagcggcc gcgtgttaca agcttgactc
     4561 cagcgtaact ggactgcaat caactcactg gctcaccttc acgggtgggc ctttcttcgg
     4621 tagaaaatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4681 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4741 tttccgaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     4801 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4861 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4921 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4981 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     5041 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     5101 caggagagcg cacgagggag cttccagggg taaacgcctg gtatctttat agtcctgtcg
     5161 ggtttcgcca cctctgactt gagcatcgat ttttgtgatg ctcgtcaggg gggcggagcc
     5221 tatggaaaaa cgccagcaac gcagaaaggc ccacccgaag gtgagccagg tgattacatt
     5281 tgggccctca tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     5341 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     5401 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     5461 cggttttttc ctgtttggtc acttaccaat gcttaatcag tgaggcacct atctcagcga
     5521 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatgc
     5581 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5641 ctccagattt atcagcaata aaccagccag ccgggagggc cgagcgcaga agtgatcctg
     5701 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5761 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5821 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5881 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gccagaagta
     5941 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6001 tgccatccgt gagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6061 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6121 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcgt aaactctcaa
     6181 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6241 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6301 caaaaaaggg aataagggcg acacggaaat gttgaatact cattttagct tccttagctc
     6361 ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa
     6421 agttggaacc tcttacgtgc cgatcaagtc aaaagcctcc ggtcggaggc ttttgacttt
     6481 ctgctatgga ggtcaggtat gatttaaatg gtcagttaga
//
