LOCUS       sequence-295419-        8750 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 8750)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8750)
  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-m4133203
FEATURES             Location/Qualifiers
     source          1..8750
                     /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             2180..2197
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2290..2411
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2301..2320)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2355..2374
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2526..2572
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2602..2738
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2783..2854
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3080..3668)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3160..3179)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4168..5377
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5214..5233
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5381..6409
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6436..6484
                     /note="synthetic polyadenylation signal"
     CDS             join(6750..6818,6819..7610)
                     /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(6968..6987)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7623..7759
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7904..7975
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7980..8026)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    8044..8092
                     /note="synthetic polyadenylation signal"
     enhancer        8173..8476
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8478..8680
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8631..8651
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(8749..8750,1..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 gatgagtgac aaaatcattc atctaaccga
     1861 tgactccttt gatacagacg tgttgaaggc cgatggtgcg atactggtag atttctgggc
     1921 tgaatggtgt ggcccgtgca agatgatcgc acccattctg gacgagatcg ccgacgagta
     1981 tcaaggaaaa ttaactgtcg ctaagcttaa tatagatcag aacccaggga cggcacctaa
     2041 atacggcatc aggggaatac ctaccctcct gctctttaag aatggggaag ttgccgcgac
     2101 aaaagtggga gccctgtcta agggccagct aaaagagttc ttggacgcta acctggcagg
     2161 ttcaggctcg gggcacatgc accatcacca tcaccatagc agcgggctcg tgccacgcgg
     2221 ctaaggatcc gcaggcctct gctagcttga ctgactgaga tacagcgtac cttcagctca
     2281 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     2341 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     2401 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     2461 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tattggccca tctctatcgg
     2521 tatcgtagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgtgcccctc
     2581 gggccggatt gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc
     2641 gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac
     2701 ttccatacgt gtcctcctta ccagaaattt atccttaagg tcgtcagcta tcctgcaggc
     2761 gatctctcga tttcgatcaa gacattcctt taatggtctt ttctggacac cactaggggt
     2821 cagaagtagt tcatcaaact ttcttccctc cctaatctca ttggttacct tgggctatcg
     2881 aaacttaatt aaccagtcaa gtcagctact tggcgagatc gacttgtctg ggtttcgact
     2941 acgctcagaa ttgcgtcagt caagttcgat ctggtccttg ctattgcacc cgttctccga
     3001 ttacgagttt catttaaatc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3061 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3121 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3181 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3241 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3301 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3361 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3421 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3481 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3541 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3601 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3661 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3721 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3781 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3841 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3901 ttgcatttaa atttccgaac tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc
     3961 cgatccatct tgcaggctac ctctcgaacg aactatcgca agtctcttgg ccggccttgc
     4021 gccttggcta ttgcttggca gcgcctatcg ccaggtatta ctccaatccc gaatatccga
     4081 gatcgggatc acccgagaga agttcaacct acatcctcaa tcccgatcta tccgagatcc
     4141 gaggaatatc gaaatcgggg cgcgcctggc ctccgcgccg ggttttggcg cctcccgcgg
     4201 gcgcccccct cgtcacggcg agcgctgcca cgtcagacga agggcgcagg agcgtcctga
     4261 tccttccgcc cggacgctca ggacagcggc ccgctgctca taagactcgg ccttagaacc
     4321 ccagtatcag cagaaggaca ttttaggacg ggacttgggt gactctaggg cactggtttt
     4381 ctttccagag agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg
     4441 atctccgtgg ggcggtgaac gccgatgatt atataaggac gcgccgggtg tggcacagct
     4501 agttccgtcg cagccgggat ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca
     4561 cttggtgagt agcgggctgc tgggctggcc ggggctttcg tggccgccgg gccgctcggt
     4621 gggacggaag cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg
     4681 ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg
     4741 gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg
     4801 gcaagaaccc aaggtcttga gcccttcgct aatgcgggaa agctcttatt cgggtgagat
     4861 gggctgggca ccatctgggg accctgacgt gaagtttgtc actgactgga gaactcggtt
     4921 tgtcgtctgt tgcgggggcg gcagttatgg cggtgccgtt gggcagtgca cccgtacctt
     4981 tgggagcgcg cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg
     5041 gtggggccac ctgccggtag gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc
     5101 gggcctaggg taggctctcc tgaatcgaca ggcgccggac ctctggtgag gggagggata
     5161 agtgaggcgt cagtttcttt ggtcggtttt atgtacctat cttcttaagt agctgaagct
     5221 ccggttttga actatgcgct cggggttggc gagtgtgttt tgtgaagttt tttaggcacc
     5281 ttttgaaatg taatcatttg ggtcaatatg taattttcag tgttagactt gtaaattgtc
     5341 cgctaaattc tggccgtttt tggctttttt gttagacaac atgggtaaaa agcctgaact
     5401 caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat
     5461 gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata
     5521 tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca
     5581 ctttgcatcc gccgcgctcc cgattccgga agtgcttgac attggggagt tcagcgagag
     5641 cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac
     5701 cgaactgccc gctgttctgc agccggtcgc ggaggcaatg gatgccatcg ctgccgccga
     5761 tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac
     5821 atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat
     5881 ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga
     5941 ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac
     6001 ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca
     6061 atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac
     6121 gcgctacttc gagcggaggc atccggagct tgcaggatcg ccccggctcc gggcgtatat
     6181 gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc
     6241 agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg
     6301 tacacaaatc gcccgcagaa gcgccgccgt ctggaccgat ggctgtgtag aagtactcgc
     6361 cgatagtgga aaccgacgcc ccagcactcg tccgagggca aaggaataag ctagtatgta
     6421 agcctagtct tagataataa aatcgctatc catcgaagat ggatgtgtgt tggttttttg
     6481 tgtgtgtaac gctaggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga
     6541 cgatccatat cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc
     6601 caatcgtcag atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta
     6661 gatattgata gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc
     6721 aagagacagg atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta
     6781 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     6841 taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca
     6901 gcggtaagat ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta
     6961 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     7021 gccgcataca ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc
     7081 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     7141 ctgcggccaa cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc
     7201 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     7261 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac
     7321 tattaactgg cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg
     7381 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     7441 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     7501 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     7561 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag
     7621 gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
     7681 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
     7741 accagaaatt tatccttaag atcgtttaaa ctcgactctg gctctatcga atctccgtcg
     7801 tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgggcatcg aatctcgtca
     7861 gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg gaccattagc
     7921 tccacaggta tcttcttccc tctagtggtc ataacagcag cttcagctac ctctcaattc
     7981 aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt
     8041 acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta ttatctaact
     8101 gctgatcgag tgtagccaga tctagtaatc aattacgggg tcattagttc atagcccata
     8161 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     8221 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     8281 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     8341 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     8401 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     8461 catcgctatt accatgctga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     8521 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     8581 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     8641 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
     8701 cagatctttg tcgatcctac catccactcg acacacccgc cagcggccgc
//
