LOCUS       sequence-295392-        8378 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 8378)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8378)
  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-m4133183
FEATURES             Location/Qualifiers
     source          1..8378
                     /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..1849
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1918..2039
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1929..1948)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1983..2002
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2154..2200
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2230..2366
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2411..2482
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2708..3296)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2788..2807)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3796..5005
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4842..4861
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5009..6037
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6064..6112
                     /note="synthetic polyadenylation signal"
     CDS             join(6378..6446,6447..7238)
                     /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(6596..6615)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7251..7387
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7532..7603
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7608..7654)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7672..7720
                     /note="synthetic polyadenylation signal"
     enhancer        7801..8104
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8106..8308
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8259..8279
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(8377..8378,1..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 gcaccatcac catcaccatt aaggatccgc
     1861 aggcctctgc tagcttgact gactgagata cagcgtacct tcagctcaca gacatgataa
     1921 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     1981 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
     2041 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
     2101 aaagcaagta aaacctctac aaatgtggta ttggcccatc tctatcggta tcgtagcata
     2161 accccttggg gcctctaaac gggtcttgag gggttttttg tgcccctcgg gccggattgc
     2221 tatctaccgg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc
     2281 cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt
     2341 cctccttacc agaaatttat ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt
     2401 tcgatcaaga cattccttta atggtctttt ctggacacca ctaggggtca gaagtagttc
     2461 atcaaacttt cttccctccc taatctcatt ggttaccttg ggctatcgaa acttaattaa
     2521 ccagtcaagt cagctacttg gcgagatcga cttgtctggg tttcgactac gctcagaatt
     2581 gcgtcagtca agttcgatct ggtccttgct attgcacccg ttctccgatt acgagtttca
     2641 tttaaatcat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2701 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     2761 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2821 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     2881 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     2941 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3001 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3061 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3121 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3181 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3241 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3301 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3361 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3421 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3481 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcatttaaat
     3541 ttccgaactc tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg
     3601 caggctacct ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc cttggctatt
     3661 gcttggcagc gcctatcgcc aggtattact ccaatcccga atatccgaga tcgggatcac
     3721 ccgagagaag ttcaacctac atcctcaatc ccgatctatc cgagatccga ggaatatcga
     3781 aatcggggcg cgcctggcct ccgcgccggg ttttggcgcc tcccgcgggc gcccccctcg
     3841 tcacggcgag cgctgccacg tcagacgaag ggcgcaggag cgtcctgatc cttccgcccg
     3901 gacgctcagg acagcggccc gctgctcata agactcggcc ttagaacccc agtatcagca
     3961 gaaggacatt ttaggacggg acttgggtga ctctagggca ctggttttct ttccagagag
     4021 cggaacaggc gaggaaaagt agtcccttct cggcgattct gcggagggat ctccgtgggg
     4081 cggtgaacgc cgatgattat ataaggacgc gccgggtgtg gcacagctag ttccgtcgca
     4141 gccgggattt gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag
     4201 cgggctgctg ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg gacggaagcg
     4261 tgtggagaga ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc ctgaactggg
     4321 ggttgggggg agcgcagcaa aatggcggct gttcccgagt cttgaatgga agacgcttgt
     4381 gaggcgggct gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc aagaacccaa
     4441 ggtcttgagc ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg gctgggcacc
     4501 atctggggac cctgacgtga agtttgtcac tgactggaga actcggtttg tcgtctgttg
     4561 cgggggcggc agttatggcg gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg
     4621 ccctcgtcgt gtcgtgacgt cacccgttct gttggcttat aatgcagggt ggggccacct
     4681 gccggtaggt gtgcggtagg cttttctccg tcgcaggacg cagggttcgg gcctagggta
     4741 ggctctcctg aatcgacagg cgccggacct ctggtgaggg gagggataag tgaggcgtca
     4801 gtttctttgg tcggttttat gtacctatct tcttaagtag ctgaagctcc ggttttgaac
     4861 tatgcgctcg gggttggcga gtgtgttttg tgaagttttt taggcacctt ttgaaatgta
     4921 atcatttggg tcaatatgta attttcagtg ttagacttgt aaattgtccg ctaaattctg
     4981 gccgtttttg gcttttttgt tagacaacat gggtaaaaag cctgaactca ccgcgacgtc
     5041 tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga
     5101 gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt
     5161 aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatccgc
     5221 cgcgctcccg attccggaag tgcttgacat tggggagttc agcgagagcc tgacctattg
     5281 catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc
     5341 tgttctgcag ccggtcgcgg aggcaatgga tgccatcgct gccgccgatc ttagccagac
     5401 gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt
     5461 catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt
     5521 cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga
     5581 agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg
     5641 cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc
     5701 caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga
     5761 gcggaggcat ccggagcttg caggatcgcc ccggctccgg gcgtatatgc tccgcattgg
     5821 tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca
     5881 gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc
     5941 ccgcagaagc gccgccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa
     6001 ccgacgcccc agcactcgtc cgagggcaaa ggaataagct agtatgtaag cctagtctta
     6061 gataataaaa tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg tgtgtaacgc
     6121 taggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg
     6181 ttgcttggca gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat
     6241 attgtactca agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt
     6301 ctgatcggtc aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat
     6361 cagcaggagg ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     6421 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     6481 aagatcagtt gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     6541 ttgagagttt tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat
     6601 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     6661 attctcagaa tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca
     6721 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     6781 tacttctgac aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     6841 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     6901 agcgtgacac cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg
     6961 aactacttac tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg
     7021 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     7081 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     7141 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     7201 tcgctgagat aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc
     7261 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
     7321 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
     7381 tccttaagat cgtttaaact cgactctggc tctatcgaat ctccgtcgtt tcgagcttac
     7441 gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc
     7501 ttaccttttt ggcagcgatc gcggctcccg acatcttgga ccattagctc cacaggtatc
     7561 ttcttccctc tagtggtcat aacagcagct tcagctacct ctcaattcaa aaaacccctc
     7621 aagacccgtt tagaggcccc aaggggttat gctatcaatc gttgcgttac acacacaaaa
     7681 aaccaacaca catccatctt cgatggatag cgattttatt atctaactgc tgatcgagtg
     7741 tagccagatc tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     7801 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     7861 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     7921 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     7981 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     8041 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     8101 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     8161 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     8221 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     8281 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
     8341 gatcctacca tccactcgac acacccgcca gcggccgc
//
