LOCUS       sequence-295525-        8387 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 8387)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8387)
  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-m4134248
FEATURES             Location/Qualifiers
     source          1..8387
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1811..1831
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1832..1858
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1927..2048
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1938..1957)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1992..2011
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2163..2209
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2239..2375
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2420..2491
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2717..3305)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2797..2816)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3805..5014
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4851..4870
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5018..6046
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6073..6121
                     /note="synthetic polyadenylation signal"
     CDS             join(6387..6455,6456..7247)
                     /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(6605..6624)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7260..7396
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7541..7612
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7617..7663)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7681..7729
                     /note="synthetic polyadenylation signal"
     enhancer        7810..8113
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8115..8317
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8268..8288
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(8386..8387,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggcg gtgggggctt gcgtactatg taacagccag acgtcattgc gctgcggtgc
       61 atgtataaga aggccgtttc tgtgttgcaa atgctgctac gatcatgtta tttcgacttc
      121 tcacaagctt gtgctcagtg tcaatcctta tgtgtgtaat gcccccggat gcgacgtcac
      181 agatgttacc caactgtacc tgggcggcat gtcctattat tgtaagagcc acaaaccacc
      241 aatctccttc cccctgtgcg ccaacggaca ggtgtttggg ctctacaaaa acacatgtgt
      301 gggatctgac aatgtcaccg acttcaacgc tatcgccaca tgcgattgga ctaatgcagg
      361 cgactacatt ttagccaaca cctgtaccga acggctgaag ttatttgcgg ctgagactct
      421 aaaggcaact gaggaaacct tcaaactttc atatggtatc gccacggtac gtgaggtgtt
      481 gtctgatcga gaactgcatc tcagttggga ggtcgggaag cctagacctc cccttaatcg
      541 aaattacgtg ttcacaggtt atagggttac caagaacagc aaagtccaga tcggcgaata
      601 tacttttgag aagggagatt acggtgacgc cgtagtgtat cgtggcacaa caacatacaa
      661 actgaatgtt ggggattact tcgtgctgac gtcccacacc gttatgccgc tctcagctcc
      721 aactttggtt ccccaggaac attacgtgcg gattactgga ctctatccaa cgctaaacat
      781 aagcgacgag ttttccagca acgtggctaa ttaccaaaag gtcggcatgc agaagtactc
      841 cacccttcag ggtcctcctg ggactggcaa aagtcacttc gccatcggac tggcacttta
      901 ctatcccagt gcgcgcattg tatataccgc ctgctcacat gcggcagtgg acgccctgtg
      961 tgagaaggct ttaaagtatc tgccaataga caaatgctct cgcattatcc ctgccagggc
     1021 cagagtggag tgctttgata agtttaaagt gaactcgacc ctcgaacagt atgtcttctg
     1081 cacagttaac gcattgcccg aaacgaccgc cgatatagtt gtgttcgacg aaatctcaat
     1141 ggctaccaat tatgatctct ccgtcgtcaa tgcacggctg cgggccaaac actacgtcta
     1201 catcggggac cccgcgcagt taccagcacc gcgaacccta ctgactaagg gcacacttga
     1261 gccagagtac ttcaacagcg tatgtagact gatgaaaacc attggccctg atatgttcct
     1321 gggaacatgt cggaggtgcc ccgctgaaat agtggacacc gtttcagcac ttgtgtacga
     1381 caataagctc aaagcacaca aggataaatc cgcacaatgt tttaagatgt tttacaaagg
     1441 ggttattacc catgatgtgt ctagcgctat caatcgccct cagattggcg tggtgagaga
     1501 gtttttgacg cgaaatcccg cctggcgaaa ggccgtattc atcagtcctt acaactcgca
     1561 aaatgccgtc gctagtaaga ttttaggact ccctactcag acagttgata gctctcaagg
     1621 ttctgaatac gattatgtga tttttactca gactaccgaa acggcacatt cttgtaacgt
     1681 caaccgtttt aacgtggcca ttacaagggc caaggttggc atcttgtgta tcatgtcaga
     1741 ccgggacctg tatgataaac tgcaattcac atctttggag ataccgcgcc gcaatgtcgc
     1801 tacccttcag gaaaaccttt actttcagtc gtacccctat gacgtaccag attatgcgta
     1861 aggatccgca ggcctctgct agcttgactg actgagatac agcgtacctt cagctcacag
     1921 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     1981 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     2041 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     2101 aggtttttta aagcaagtaa aacctctaca aatgtggtat tggcccatct ctatcggtat
     2161 cgtagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgt gcccctcggg
     2221 ccggattgct atctaccggc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc
     2281 ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc
     2341 catacgtgtc ctccttacca gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat
     2401 ctctcgattt cgatcaagac attcctttaa tggtcttttc tggacaccac taggggtcag
     2461 aagtagttca tcaaactttc ttccctccct aatctcattg gttaccttgg gctatcgaaa
     2521 cttaattaac cagtcaagtc agctacttgg cgagatcgac ttgtctgggt ttcgactacg
     2581 ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt tctccgatta
     2641 cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2701 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2761 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2821 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2881 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2941 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3001 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     3061 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     3121 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     3181 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3241 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3301 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3361 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3421 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3481 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3541 catttaaatt tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga
     3601 tccatcttgc aggctacctc tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc
     3661 ttggctattg cttggcagcg cctatcgcca ggtattactc caatcccgaa tatccgagat
     3721 cgggatcacc cgagagaagt tcaacctaca tcctcaatcc cgatctatcc gagatccgag
     3781 gaatatcgaa atcggggcgc gcctggcctc cgcgccgggt tttggcgcct cccgcgggcg
     3841 cccccctcgt cacggcgagc gctgccacgt cagacgaagg gcgcaggagc gtcctgatcc
     3901 ttccgcccgg acgctcagga cagcggcccg ctgctcataa gactcggcct tagaacccca
     3961 gtatcagcag aaggacattt taggacggga cttgggtgac tctagggcac tggttttctt
     4021 tccagagagc ggaacaggcg aggaaaagta gtcccttctc ggcgattctg cggagggatc
     4081 tccgtggggc ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg cacagctagt
     4141 tccgtcgcag ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg atcgtcactt
     4201 ggtgagtagc gggctgctgg gctggccggg gctttcgtgg ccgccgggcc gctcggtggg
     4261 acggaagcgt gtggagagac cgccaagggc tgtagtctgg gtccgcgagc aaggttgccc
     4321 tgaactgggg gttgggggga gcgcagcaaa atggcggctg ttcccgagtc ttgaatggaa
     4381 gacgcttgtg aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg gtgggcggca
     4441 agaacccaag gtcttgagcc cttcgctaat gcgggaaagc tcttattcgg gtgagatggg
     4501 ctgggcacca tctggggacc ctgacgtgaa gtttgtcact gactggagaa ctcggtttgt
     4561 cgtctgttgc gggggcggca gttatggcgg tgccgttggg cagtgcaccc gtacctttgg
     4621 gagcgcgcgc cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata atgcagggtg
     4681 gggccacctg ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc agggttcggg
     4741 cctagggtag gctctcctga atcgacaggc gccggacctc tggtgagggg agggataagt
     4801 gaggcgtcag tttctttggt cggttttatg tacctatctt cttaagtagc tgaagctccg
     4861 gttttgaact atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt aggcaccttt
     4921 tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt tagacttgta aattgtccgc
     4981 taaattctgg ccgtttttgg cttttttgtt agacaacatg ggtaaaaagc ctgaactcac
     5041 cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca
     5101 gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt
     5161 cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt
     5221 tgcatccgcc gcgctcccga ttccggaagt gcttgacatt ggggagttca gcgagagcct
     5281 gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga
     5341 actgcccgct gttctgcagc cggtcgcgga ggcaatggat gccatcgctg ccgccgatct
     5401 tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg
     5461 gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga
     5521 cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga
     5581 ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga
     5641 caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata
     5701 cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg
     5761 ctacttcgag cggaggcatc cggagcttgc aggatcgccc cggctccggg cgtatatgct
     5821 ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc
     5881 ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac
     5941 acaaatcgcc cgcagaagcg ccgccgtctg gaccgatggc tgtgtagaag tactcgccga
     6001 tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaataagcta gtatgtaagc
     6061 ctagtcttag ataataaaat cgctatccat cgaagatgga tgtgtgttgg ttttttgtgt
     6121 gtgtaacgct aggcgcgcct ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga
     6181 tccatatcgt tgcttggcag tcagccagtc ggaatccagc ttgggaccca ggaagtccaa
     6241 tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac cgatctgttt aaacctagat
     6301 attgatagtc tgatcggtca acgtataatc gagtcctagc ttttgcaaac atctatcaag
     6361 agacaggatc agcaggaggc tttcgcatga gtattcaaca tttccgtgtc gcccttattc
     6421 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     6481 aagatgctga agatcagttg ggtgcgcgag tgggttacat cgaactggat ctcaacagcg
     6541 gtaagatcct tgagagtttt cgccccgaag aacgctttcc aatgatgagc acttttaaag
     6601 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     6661 gcatacacta ttctcagaat gacttggttg agtattcacc agtcacagaa aagcatctta
     6721 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     6781 cggccaactt acttctgaca acgattggag gaccgaagga gctaaccgct tttttgcaca
     6841 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     6901 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaaccttg cgtaaactat
     6961 taactggcga actacttact ctagcttccc ggcaacagtt gatagactgg atggaggcgg
     7021 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     7081 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     7141 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     7201 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaaccg attctaggtg
     7261 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
     7321 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
     7381 agaaatttat ccttaagatc gtttaaactc gactctggct ctatcgaatc tccgtcgttt
     7441 cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct
     7501 atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac cattagctcc
     7561 acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc tcaattcaaa
     7621 aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg ttgcgttaca
     7681 cacacaaaaa accaacacac atccatcttc gatggatagc gattttatta tctaactgct
     7741 gatcgagtgt agccagatct agtaatcaat tacggggtca ttagttcata gcccatatat
     7801 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     7861 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     7921 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     7981 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     8041 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     8101 cgctattacc atgctgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     8161 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     8221 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     8281 taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatcag
     8341 atctttgtcg atcctaccat ccactcgaca cacccgccag cggccgc
//
