LOCUS       sequence-298430-        6503 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Yeast Expression plasmid for SARS-CoV-2 helicase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6503)
  TITLE     SARS-CoV-2 protein Yeast expression constructs, tagged, for protein 
            expression and/or purification
REFERENCE   2  (bases 1 to 6503)
  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-m4046656
FEATURES             Location/Qualifiers
     source          1..6503
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(2261..2279)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             2630..3430
                     /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
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRCGQQN"
     rep_origin      3513..4495
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     terminator      4573..4602
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      4624..5211
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5223..5250)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(5274..5465)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5466..6257,6258..6326))
                     /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     6089..6108
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6327..6429)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(6430..6461)
                     /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 caataaattc ttgaaaattt
      241 ttttttttga tttttttctc tttcgatgac ctcccattga tatttaagtt aataaacggt
      301 cttcaatttc tcaagtttca gtttcatttt tcttgttcta ttacaacttt ttttacttct
      361 tgctcattag aaagaaagca tagcaatcta atctaagttt taattacaaa atggcagtcg
      421 gcgcttgtgt attatgcaac tcccagacgt cgttgagatg tggtgcttgt attcgtagac
      481 ctttcctttg ctgtaaatgt tgctatgacc acgttatctc aacttctcat aagctagttc
      541 tgagcgtcaa tccatacgtg tgtaacgccc ccgggtgtga tgttacagac gttacccaat
      601 tgtacttggg aggtatgtct tactattgta agtcccacaa gccacctata tcttttccat
      661 tatgcgctaa cggtcaagtc ttcggtttgt acaaaaatac ctgtgttggt agtgataatg
      721 ttaccgactt taacgccatt gctacttgtg attggactaa cgcgggtgat tatatcttgg
      781 ctaacacttg taccgagagg cttaagttgt tcgccgctga aaccttaaag gctacggaag
      841 aaacattcaa attgagttac ggtatcgcta ctgtcagaga agttctatct gatagagaat
      901 tgcatctctc ctgggaggtg ggtaagccaa gaccaccgtt gaaccggaat tacgtcttta
      961 ctggatacag ggttactaag aactctaagg ttcaaattgg ggaatatacc tttgaaaagg
     1021 gcgactacgg tgacgccgta gtctatagag gtaccaccac ttacaaattg aacgtcggtg
     1081 actatttcgt tttaaccagc cacacagtga tgccactgtc agcgccaaca ttagttccgc
     1141 aagagcatta cgttagaatt actggtttgt atccaactct aaatatctct gatgaatttt
     1201 cctccaatgt ggctaactac caaaaagtag gtatgcaaaa gtactctacc ttgcaaggtc
     1261 cacctggtac tgggaagtcg cacttcgcta taggtttggc actgtactac ccatctgcta
     1321 gaattgttta cacggcatgt tctcatgctg ctgtagacgc cttgtgtgaa aaggccttga
     1381 aatatttacc cattgataaa tgttcaagaa ttatcccagc acgagctcgt gttgaatgtt
     1441 tcgacaagtt taaggtcaac agcacactag aacaatacgt cttctgcact gttaatgctt
     1501 tgccagaaac gactgctgat atcgtcgtct ttgatgaaat ctcgatggcc actaactatg
     1561 acttatccgt agttaatgct agacttcgtg caaagcacta cgtctatatt ggtgatccag
     1621 ctcagctgcc tgcaccaaga actttgttaa ccaagggtac cttggaacca gaatacttca
     1681 actctgtgtg tcgcctgatg aagacaatcg gccctgatat gttcctaggt acatgtagac
     1741 gttgtccagc tgaaattgtg gacacggtca gtgccttggt ttacgataac aaattaaagg
     1801 cgcataagga caaaagcgct caatgtttca agatgtttta caagggtgtc attactcacg
     1861 atgttagttc ggccatcaat agaccccaaa taggtgttgt tagagaattt ttgacaagga
     1921 acccagcatg gagaaaggct gttttcatat ccccatacaa ttctcaaaac gcggtagcct
     1981 caaaaatctt gggtcttccc actcagaccg ttgattcttc ccaaggaagt gaatatgact
     2041 acgtgatttt cactcaaacc accgagactg cccactcttg caacgtcaat cgtttcaacg
     2101 ttgctattac gagagccaag gttggcatct tgtgtatcat gtctgataga gacttgtacg
     2161 ataaattaca gtttacatcc ttagaaatac caagacgaaa cgtcgctact ttgcaataaa
     2221 caggcccctt ttcctttgtc gatatcatgt aattagttat gtcacgctta cattcacgcc
     2281 ctcctcccac atccgctcta accgaaaagg aaggagttag acaacctgaa gtctaggtcc
     2341 ctatttattt tttttaatag ttatgttagt attaagaacg ttatttatat ttcaaatttt
     2401 tctttttttt ctgtacaaag cggccgccga accactagtg cactgcagta cacggtttcc
     2461 ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag
     2521 acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt
     2581 cttaacccaa ctgcacagaa caaaaacgtg caggaaacga agataaatca tgtcgaaagc
     2641 tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat
     2701 catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt
     2761 actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat
     2821 cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta
     2881 caatttttta ctcttcgagg acagaaaatt tgctgacatt ggtaatacag tcaaattgca
     2941 gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt
     3001 ggtgggccca ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc
     3061 tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata
     3121 tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc
     3181 tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt
     3241 gggtttagat gacaagggag atgcattggg tcaacagtat agaaccgtgg atgatgtggt
     3301 ttctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc
     3361 taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gatgcggcca
     3421 gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac aaattagagc
     3481 ttcaatttaa ttatatcagt tattacccga gtaacgaagc atctgtgctt cattttgtag
     3541 aacaaaaatg caacgcgaga gcgctaattt twcaaacaaa gaatctgagc tgcattttta
     3601 cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt gcttcatttt
     3661 tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc tgagctgcat
     3721 ttttacagaa cagaaatgca acgcgagagc gctattttac caacaaagaa tctatacttc
     3781 ttttttgttc tacaaaaatg catcccgaga gcgctatttt tctaacaaag catcttagat
     3841 tacttttttt ctcctttgtg cgctctataa tgcagtctct tgataacttt ttgcactgta
     3901 ggtccgttaa ggttagaaga aggctacttt ggtgtctatt ttctcttcca taaaaaaagc
     3961 ctgactccac ttcccgcgtt tactgattac tagcgaagct gcgggtgcat tttttcaaga
     4021 taaaggcatc cccgattata ttctataccg atgtggattg cgcatacttt gtgaacagaa
     4081 agtgatagcg ttgatgattc ttcattggtc agaaaattat gaacggtttc ttctattttg
     4141 tctctatata ctacgtatag gaaatgttta cattttcgta ttgttttcga ttcactctat
     4201 gaatagttct tactacaatt tttttgtcta aagagtaata ctagagataa acataaaaaa
     4261 tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag gtggatgggt aggttatata
     4321 gggatatagc acagagatat atagcaaaga gatacttttg agcaatgttt gtggaagcgg
     4381 tattcgcaat attttagtag ctcgttacag tccggtgcgt ttttggtttt ttgaaagtgc
     4441 gtcatcagag cgcttttggt tttcaaaagc gctctgaagt tcctatactt tctagctaga
     4501 gaataggaac ttcccgagcg gccgcgtgtt acaagcttga ctccagcgta actggactgc
     4561 aatcaactca ctggctcacc ttcacgggtg ggcctttctt cggtagaaaa tcaaaggatc
     4621 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4681 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga ggtaactggc
     4741 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4801 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4861 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4921 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4981 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5041 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5101 gagcttccag gggtaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5161 cttgagcatc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5221 aacgcagaaa ggcccacccg aaggtgagcc aggtgattac atttgggccc tcatcagagg
     5281 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
     5341 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
     5401 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
     5461 gtcacttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5521 tccatagttg cctgactccc cgtcgtgtag ataactacga tgcgggaggg cttaccatct
     5581 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5641 ataaaccagc cagccgggag ggccgagcgc agaagtgatc ctgcaacttt atccgcctcc
     5701 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5761 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5821 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5881 aaagcggtta gctccttcgg tcctccgatc gttgccagaa gtaagttggc cgcagtgtta
     5941 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtgagatgc
     6001 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6061 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6121 gtgctcatca ttggaaaacg ttcttcgggg cgtaaactct caaggatctt accgctgttg
     6181 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6241 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6301 gcgacacgga aatgttgaat actcatttta gcttccttag ctcctgaaaa tctcgataac
     6361 tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg
     6421 tgccgatcaa gtcaaaagcc tccggtcgga ggcttttgac tttctgctat ggaggtcagg
     6481 tatgatttaa atggtcagtt aga
//
