LOCUS       sequence-300503-        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"
     CDS             complement(302..1105)
                     /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"
     primer_bind     1456..1474
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(3332..3745)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      3791..3822
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        3823..3925
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(3926..3994,3995..4786)
                     /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     complement(4144..4163)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             4787..4978
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      5002..5029
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(5041..5628)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5650..5679)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      join(5757..6520,1..219)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 gttctgtaaa aatgcagctc agattctttg tttgaaaaat tagcgctctc gcgttgcatt
       61 tttgttttac aaaaatgaag cacagattct tcgttggtaa aatagcgctt tcgcgttgca
      121 tttctgttct gtaaaaatgc agctcagatt ctttgtttga aaaattagcg ctctcgcgtt
      181 gcatttttgt tctacaaaat gaagcacaga tgcttcgtta ctcgggtaat aactgatata
      241 attaaattga agctctaatt tgtgagttta gtatacatgc atttacttat aatacagttt
      301 tttagttttg ctggccgcat cttctcaaat atgcttccca gcctgctttt ctgtaacgtt
      361 caccctctac cttagcatcc cttccctttg caaatagtcc tcttccaaca ataataatgt
      421 cagatcctgt agaaaccaca tcatccacgg ttctatactg ttgacccaat gcatctccct
      481 tgtcatctaa acccacaccg ggtgtcataa tcaaccaatc gtaaccttca tctcttccac
      541 ccatgtctct ttgagcaata aagccgataa caaaatcttt gtcgctcttc gcaatgtcaa
      601 cagtaccctt agtatattct ccagtagata gggagccctt gcatgacaat tctgctaaca
      661 tcaaaaggcc tctaggttcc tttgttactt cttctgccgc ctgcttcaaa ccgctaacaa
      721 tacctgggcc caccacaccg tgtgcattcg taatgtctgc ccattctgct attctgtata
      781 cacccgcaga gtactgcaat ttgactgtat taccaatgtc agcaaatttt ctgtcctcga
      841 agagtaaaaa attgtacttg gcggataatg cctttagcgg cttaactgtg ccctccatgg
      901 aaaaatcagt caagatatcc acatgtgttt ttagtaaaca aattttggga cctaatgctt
      961 caactaactc cagtaattcc ttggtggtac gaacatccaa tgaagcacac aagtttgttt
     1021 gcttttcgtg catgatatta aatagcttgg cagcaacagg actaggatga gtagcagcac
     1081 gttccttata tgtagctttc gacatgattt atcttcgttt cctgcacgtt tttgttctgt
     1141 gcagttgggt taagaatact gggcaatttc atgtttcttc aacactacat atgcgtatat
     1201 ataccaatct aagtctgtgc tccttccttc gttcttcctt ctgttcggag attaccgaat
     1261 caaaaaaatt tcaaggaaac cgtgtactgc agtgcactag tggttcggcg gccgctttgt
     1321 acagaaaaaa aagaaaaatt tgaaatataa ataacgttct taatactaac ataactatta
     1381 aaaaaaataa atagggacct agacttcagg ttgtctaact ccttcctttt cggttagagc
     1441 ggatgtggga ggagggcgtg aatgtaagcg tgacataact aattacatga tatcgacaaa
     1501 ggaaaagggg cctgtttata aagttgccac attcctacgt ggaatttcaa gacttgtaaa
     1561 ttgcaacttg tcataaaggt ctctatcaga cattatgcaa agtatgccta cttttgctct
     1621 ggtaatagca acattaaatc tgtttacatt acaagagtga gctgtttcag tggtttgagt
     1681 gaatatgaca tagtcatatt ctgagccctg tgatgaatca acagtttgag ttggtagtcc
     1741 caaaatcttt gaggctacag cattctgtga attataaggt gaaataaaga cagcttttct
     1801 ccaagcaggg ttacgtgtaa ggaattctct taccacgcct atttgtggcc tgttaattgc
     1861 agatgaaaca tcatgcgtga taacaccctt ataaaacatt ttaaagcatt gagctgattt
     1921 gtctttatgt gctttaagct tattatcata aaccaaagca ctcacagtgt caacaatttc
     1981 agcaggacaa cgccgacaag ttccgaggaa catgtctgga cctatagttt tcataagtct
     2041 acacactgaa ttgaaatatt ctggttctag tgtgccctta gttagcaatg tgcgtggtgc
     2101 aggtaattga gcagggtcgc caatgtacac atagtgctta gcacgtaatc tggcattgac
     2161 aacactcaaa tcataatttg tggccattga aatttcatca aagacaacta tatctgctgt
     2221 cgtctcaggc aatgcattta cagtacaaaa gacatactgt tctaatgttg aattcacttt
     2281 gaatttatca aaacactcta cacgagcacg tgcaggtata attctactac atttatctat
     2341 aggcaaatat tttaatgcct tctcacatag tgcatcaaca gcggcatgag agcaagctgt
     2401 atacactatg cgagcagaag ggtagtagag agctaggcca atagcaaaat gactcttacc
     2461 agtaccaggt ggtccctgga gtgtagaata cttttgcata ccaacctttt gataatttgc
     2521 aacattgcta gaaaactcat ctgagatatt gagtgttggg tataagccag taattctaac
     2581 atagtgctct tgtggcacta gtgtaggtgc acttaatggc attactgtat gtgatgtcag
     2641 cacaaaataa tcaccaacat ttaatttgta agttgttgta cctcggtaaa caacagcatc
     2701 accatagtca cctttttcaa aggtgtactc tcctatttgt actttactgt ttttagttac
     2761 acgataacca gtaaagacat aatttcggtt aagtggtggt ctaggtttac caacttccca
     2821 tgaaagatgt aattctctgt cagacagcac ttcacgtaca gtagcaatac cataagacag
     2881 tttaaatgtc tcctcagtag ctttgagcgt ttctgctgca aaaagcttga gtctttcagt
     2941 acaggtgtta gctaaaatgt aatcaccagc atttgtccag tcacatgttg caattgcatt
     3001 aaagtcagta acattatcgc taccaacaca tgtattttta tataaaccaa aaacttgtcc
     3061 attagcacac aatggaaaac taatgggtgg tttatgtgat ttacaataat agctcatacc
     3121 tcctaagtaa agttgagtca catctgtgac atcacaacct ggagcattgc aaacatacgg
     3181 attaacagac aagactaatt tatgtgatgt tgatatgaca tggtcgtaac agcatttaca
     3241 acataagaat ggtctacgta tgcaagcacc acatcttaat gaagtctgtg aattgcaaag
     3301 aacacaagcc ccaaccattt tgtaattaaa acttagatta gattgctatg ctttctttct
     3361 aatgagcaag aagtaaaaaa agttgtaata gaacaagaaa aatgaaactg aaacttgaga
     3421 aattgaagac cgtttattaa cttaaatatc aatgggaggt catcgaaaga gaaaaaaatc
     3481 aaaaaaaaaa attttcaaga aaaagaaacg tgataaaaat ttttattgcc tttttcgacg
     3541 aagaaaaaga aacgaggcgg tctctttttt cttttccaaa cctttagtac gggtaattaa
     3601 cgacacccta gaggaagaaa gaggggaaat ttagtatgct gtgcttgggt gttttgaagt
     3661 ggtacggcga tgcgcggagt ccgagaaaat ctggaagagt aaaaaaggag tagaaacatt
     3721 ttgaagctat ggtgtgtggg gcggccgctc taactgacca tttaaatcat acctgacctc
     3781 catagcagaa agtcaaaagc ctccgaccgg aggcttttga cttgatcggc acgtaagagg
     3841 ttccaacttt caccataatg aaataagatc actaccgggc gtattttttg agttatcgag
     3901 attttcagga gctaaggaag ctaaaatgag tattcaacat ttccgtgtcg cccttattcc
     3961 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4021 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4081 taagatcctt gagagtttac gccccgaaga acgttttcca atgatgagca cttttaaagt
     4141 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4201 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatctcac
     4261 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4321 ggccaactta cttctggcaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4381 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4441 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     4501 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4561 taaagttgca ggatcacttc tgcgctcggc cctcccggct ggctggttta ttgctgataa
     4621 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4681 gccctcccgc atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4741 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaagtga ccaaacagga
     4801 aaaaaccgcc cttaacatgg cccgctttat cagaagccag acattaacgc ttctggagaa
     4861 actcaacgag ctggacgcgg atgaacaggc agacatctgt gaatcgcttc acgaccacgc
     4921 tgatgagctt taccgcagct gcctcgcgcg tttcggtgat gacggtgaaa acctctgatg
     4981 agggcccaaa tgtaatcacc tggctcacct tcgggtgggc ctttctgcgt tgctggcgtt
     5041 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgatgctcaa gtcagaggtg
     5101 gcgaaacccg acaggactat aaagatacca ggcgtttacc cctggaagct ccctcgtgcg
     5161 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5221 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5281 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5341 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5401 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5461 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5521 ctcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5581 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5641 attttctacc gaagaaaggc ccacccgtga aggtgagcca gtgagttgat tgcagtccag
     5701 ttacgctgga gtcaagcttg taacacgcgg ccgctcggga agttcctatt ctctagctag
     5761 aaagtatagg aacttcagag cgcttttgaa aaccaaaagc gctctgatga cgcactttca
     5821 aaaaaccaaa aacgcaccgg actgtaacga gctactaaaa tattgcgaat accgcttcca
     5881 caaacattgc tcaaaagtat ctctttgcta tatatctctg tgctatatcc ctatataacc
     5941 tacccatcca cctttcgctc cttgaacttg catctaaact cgacctctac attttttatg
     6001 tttatctcta gtattactct ttagacaaaa aaattgtagt aagaactatt catagagtga
     6061 atcgaaaaca atacgaaaat gtaaacattt cctatacgta gtatatagag acaaaataga
     6121 agaaaccgtt cataattttc tgaccaatga agaatcatca acgctatcac tttctgttca
     6181 caaagtatgc gcaatccaca tcggtataga atataatcgg ggatgccttt atcttgaaaa
     6241 aatgcacccg cagcttcgct agtaatcagt aaacgcggga agtggagtca ggcttttttt
     6301 atggaagaga aaatagacac caaagtagcc ttcttctaac cttaacggac ctacagtgca
     6361 aaaagttatc aagagactgc attatagagc gcacaaagga gaaaaaaagt aatctaagat
     6421 gctttgttag aaaaatagcg ctctcgggat gcatttttgt agaacaaaaa agaagtatag
     6481 attctttgtt ggtaaaatag cgctctcgcg ttgcatttct
//
