LOCUS       sequence-295350-        8744 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 helicase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8744)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8744)
  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-m4134094
FEATURES             Location/Qualifiers
     source          1..8744
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1805..1825
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2174..2191
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2284..2405
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2295..2314)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2349..2368
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2520..2566
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2596..2732
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2777..2848
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3074..3662)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3154..3173)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4162..5371
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5208..5227
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5375..6403
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6430..6478
                     /note="synthetic polyadenylation signal"
     CDS             join(6744..6812,6813..7604)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6962..6981)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7617..7753
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7898..7969
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7974..8020)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    8038..8086
                     /note="synthetic polyadenylation signal"
     enhancer        8167..8470
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8472..8674
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8625..8645
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(8743..8744,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggtt ggggcttgtg ttctttgcaa ttcacagact tcattaagat gtggtgcttg
       61 catacgtaga ccattcttat gttgtaaatg ctgttacgac catgtcatat caacatcaca
      121 taaattagtc ttgtctgtta atccgtatgt ttgcaatgct ccaggttgtg atgtcacaga
      181 tgtgactcaa ctttacttag gaggtatgag ctattattgt aaatcacata aaccacccat
      241 tagttttcca ttgtgtgcta atggacaagt ttttggttta tataaaaata catgtgttgg
      301 tagcgataat gttactgact ttaatgcaat tgcaacatgt gactggacaa atgctggtga
      361 ttacatttta gctaacacct gtactgaaag actcaagctt tttgcagcag aaacgctcaa
      421 agctactgag gagacattta aactgtctta tggtattgct actgtacgtg aagtgctgtc
      481 tgacagagaa ttacatcttt catgggaagt tggtaaacct agaccaccac ttaaccgaaa
      541 ttatgtcttt actggttatc gtgtaactaa aaacagtaaa gtacaaatag gagagtacac
      601 ctttgaaaaa ggtgactatg gtgatgctgt tgtttaccga ggtacaacaa cttacaaatt
      661 aaatgttggt gattattttg tgctgacatc acatacagta atgccattaa gtgcacctac
      721 actagtgcca caagagcact atgttagaat tactggctta tacccaacac tcaatatctc
      781 agatgagttt tctagcaatg ttgcaaatta tcaaaaggtt ggtatgcaaa agtattctac
      841 actccaggga ccacctggta ctggtaagag tcattttgct attggcctag ctctctacta
      901 cccttctgct cgcatagtgt atacagcttg ctctcatgcc gctgttgatg cactatgtga
      961 gaaggcatta aaatatttgc ctatagataa atgtagtaga attatacctg cacgtgctcg
     1021 tgtagagtgt tttgataaat tcaaagtgaa ttcaacatta gaacagtatg tcttttgtac
     1081 tgtaaatgca ttgcctgaga cgacagcaga tatagttgtc tttgatgaaa tttcaatggc
     1141 cacaaattat gatttgagtg ttgtcaatgc cagattacgt gctaagcact atgtgtacat
     1201 tggcgaccct gctcaattac ctgcaccacg cacattgcta actaagggca cactagaacc
     1261 agaatatttc aattcagtgt gtagacttat gaaaactata ggtccagaca tgttcctcgg
     1321 aacttgtcgg cgttgtcctg ctgaaattgt tgacactgtg agtgctttgg tttatgataa
     1381 taagcttaaa gcacataaag acaaatcagc tcaatgcttt aaaatgtttt ataagggtgt
     1441 tatcacgcat gatgtttcat ctgcaattaa caggccacaa ataggcgtgg taagagaatt
     1501 ccttacacgt aaccctgctt ggagaaaagc tgtctttatt tcaccttata attcacagaa
     1561 tgctgtagcc tcaaagattt tgggactacc aactcaaact gttgattcat cacagggctc
     1621 agaatatgac tatgtcatat tcactcaaac cactgaaaca gctcactctt gtaatgtaaa
     1681 cagatttaat gttgctatta ccagagcaaa agtaggcata ctttgcataa tgtctgatag
     1741 agacctttat gacaagttgc aatttacaag tcttgaaatt ccacgtagga atgtggcaac
     1801 tttagaaaac ctttactttc agtcgatgag tgacaaaatc attcatctaa ccgatgactc
     1861 ctttgataca gacgtgttga aggccgatgg tgcgatactg gtagatttct gggctgaatg
     1921 gtgtggcccg tgcaagatga tcgcacccat tctggacgag atcgccgacg agtatcaagg
     1981 aaaattaact gtcgctaagc ttaatataga tcagaaccca gggacggcac ctaaatacgg
     2041 catcagggga atacctaccc tcctgctctt taagaatggg gaagttgccg cgacaaaagt
     2101 gggagccctg tctaagggcc agctaaaaga gttcttggac gctaacctgg caggttcagg
     2161 ctcggggcac atgcaccatc accatcacca tagcagcggg ctcgtgccac gcggctaagg
     2221 atccgcaggc ctctgctagc ttgactgact gagatacagc gtaccttcag ctcacagaca
     2281 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     2341 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     2401 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     2461 ttttttaaag caagtaaaac ctctacaaat gtggtattgg cccatctcta tcggtatcgt
     2521 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgtgcc cctcgggccg
     2581 gattgctatc taccggcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta
     2641 tactcccaca tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat
     2701 acgtgtcctc cttaccagaa atttatcctt aaggtcgtca gctatcctgc aggcgatctc
     2761 tcgatttcga tcaagacatt cctttaatgg tcttttctgg acaccactag gggtcagaag
     2821 tagttcatca aactttcttc cctccctaat ctcattggtt accttgggct atcgaaactt
     2881 aattaaccag tcaagtcagc tacttggcga gatcgacttg tctgggtttc gactacgctc
     2941 agaattgcgt cagtcaagtt cgatctggtc cttgctattg cacccgttct ccgattacga
     3001 gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3061 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3121 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3181 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3241 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3301 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3361 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     3421 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     3481 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     3541 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3601 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3661 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3721 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3781 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     3841 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcat
     3901 ttaaatttcc gaactctcca aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc
     3961 atcttgcagg ctacctctcg aacgaactat cgcaagtctc ttggccggcc ttgcgccttg
     4021 gctattgctt ggcagcgcct atcgccaggt attactccaa tcccgaatat ccgagatcgg
     4081 gatcacccga gagaagttca acctacatcc tcaatcccga tctatccgag atccgaggaa
     4141 tatcgaaatc ggggcgcgcc tggcctccgc gccgggtttt ggcgcctccc gcgggcgccc
     4201 ccctcgtcac ggcgagcgct gccacgtcag acgaagggcg caggagcgtc ctgatccttc
     4261 cgcccggacg ctcaggacag cggcccgctg ctcataagac tcggccttag aaccccagta
     4321 tcagcagaag gacattttag gacgggactt gggtgactct agggcactgg ttttctttcc
     4381 agagagcgga acaggcgagg aaaagtagtc ccttctcggc gattctgcgg agggatctcc
     4441 gtggggcggt gaacgccgat gattatataa ggacgcgccg ggtgtggcac agctagttcc
     4501 gtcgcagccg ggatttgggt cgcggttctt gtttgtggat cgctgtgatc gtcacttggt
     4561 gagtagcggg ctgctgggct ggccggggct ttcgtggccg ccgggccgct cggtgggacg
     4621 gaagcgtgtg gagagaccgc caagggctgt agtctgggtc cgcgagcaag gttgccctga
     4681 actgggggtt ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg aatggaagac
     4741 gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga
     4801 acccaaggtc ttgagccctt cgctaatgcg ggaaagctct tattcgggtg agatgggctg
     4861 ggcaccatct ggggaccctg acgtgaagtt tgtcactgac tggagaactc ggtttgtcgt
     4921 ctgttgcggg ggcggcagtt atggcggtgc cgttgggcag tgcacccgta cctttgggag
     4981 cgcgcgccct cgtcgtgtcg tgacgtcacc cgttctgttg gcttataatg cagggtgggg
     5041 ccacctgccg gtaggtgtgc ggtaggcttt tctccgtcgc aggacgcagg gttcgggcct
     5101 agggtaggct ctcctgaatc gacaggcgcc ggacctctgg tgaggggagg gataagtgag
     5161 gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt aagtagctga agctccggtt
     5221 ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa gttttttagg caccttttga
     5281 aatgtaatca tttgggtcaa tatgtaattt tcagtgttag acttgtaaat tgtccgctaa
     5341 attctggccg tttttggctt ttttgttaga caacatgggt aaaaagcctg aactcaccgc
     5401 gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct
     5461 ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct
     5521 gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc
     5581 atccgccgcg ctcccgattc cggaagtgct tgacattggg gagttcagcg agagcctgac
     5641 ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact
     5701 gcccgctgtt ctgcagccgg tcgcggaggc aatggatgcc atcgctgccg ccgatcttag
     5761 ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg
     5821 tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga
     5881 caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg
     5941 ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa
     6001 tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga
     6061 ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta
     6121 cttcgagcgg aggcatccgg agcttgcagg atcgccccgg ctccgggcgt atatgctccg
     6181 cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg
     6241 ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca
     6301 aatcgcccgc agaagcgccg ccgtctggac cgatggctgt gtagaagtac tcgccgatag
     6361 tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa taagctagta tgtaagccta
     6421 gtcttagata ataaaatcgc tatccatcga agatggatgt gtgttggttt tttgtgtgtg
     6481 taacgctagg cgcgcctggt gtaccgagaa cgatcctctc agtgcgagtc tcgacgatcc
     6541 atatcgttgc ttggcagtca gccagtcgga atccagcttg ggacccagga agtccaatcg
     6601 tcagatattg tactcaagcc tggtcacggc agcgtaccga tctgtttaaa cctagatatt
     6661 gatagtctga tcggtcaacg tataatcgag tcctagcttt tgcaaacatc tatcaagaga
     6721 caggatcagc aggaggcttt cgcatgagta ttcaacattt ccgtgtcgcc cttattccct
     6781 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     6841 atgctgaaga tcagttgggt gcgcgagtgg gttacatcga actggatctc aacagcggta
     6901 agatccttga gagttttcgc cccgaagaac gctttccaat gatgagcact tttaaagttc
     6961 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     7021 tacactattc tcagaatgac ttggttgagt attcaccagt cacagaaaag catcttacgg
     7081 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     7141 ccaacttact tctgacaacg attggaggac cgaaggagct aaccgctttt ttgcacaaca
     7201 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     7261 acgacgagcg tgacaccacg atgcctgtag caatggcaac aaccttgcgt aaactattaa
     7321 ctggcgaact acttactcta gcttcccggc aacagttgat agactggatg gaggcggata
     7381 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     7441 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     7501 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     7561 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaaccgatt ctaggtgcat
     7621 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
     7681 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
     7741 aatttatcct taagatcgtt taaactcgac tctggctcta tcgaatctcc gtcgtttcga
     7801 gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc
     7861 gtcagcttac ctttttggca gcgatcgcgg ctcccgacat cttggaccat tagctccaca
     7921 ggtatcttct tccctctagt ggtcataaca gcagcttcag ctacctctca attcaaaaaa
     7981 cccctcaaga cccgtttaga ggccccaagg ggttatgcta tcaatcgttg cgttacacac
     8041 acaaaaaacc aacacacatc catcttcgat ggatagcgat tttattatct aactgctgat
     8101 cgagtgtagc cagatctagt aatcaattac ggggtcatta gttcatagcc catatatgga
     8161 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     8221 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     8281 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     8341 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     8401 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     8461 tattaccatg ctgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     8521 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     8581 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     8641 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatcagatc
     8701 tttgtcgatc ctaccatcca ctcgacacac ccgccagcgg ccgc
//
