LOCUS       sequence-295851-        8909 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 8909)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8909)
  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-m4133459
FEATURES             Location/Qualifiers
     source          1..8909
                     /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             1859..1936
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             1964..1984
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    2449..2570
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2460..2479)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2514..2533
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2685..2731
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2761..2897
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2942..3013
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3239..3827)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3319..3338)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4327..5536
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5373..5392
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5540..6568
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6595..6643
                     /note="synthetic polyadenylation signal"
     CDS             join(6909..6977,6978..7769)
                     /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(7127..7146)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7782..7918
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    8063..8134
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(8139..8185)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    8203..8251
                     /note="synthetic polyadenylation signal"
     enhancer        8332..8635
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8637..8839
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8790..8810
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(8908..8909,1..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 agtcgggacg gcgaatcccg gggctgataa atccctggaa
     1861 aaggagatgg aagaagaact ttattgcagt gtctgcggct aaccggttca aaaagatcag
     1921 ttcaagcggc gccctagatt acgacattcc tacgaccgcc tccgagaatc tctatttcca
     1981 gggtgaattt ggcttggcac aacatgacga ggccgtcgat aacaagttta ataaagaaca
     2041 gcagaacgct ttctatgaga tccttcactt accaaatctg aacgaagagc aacgcaacgc
     2101 tttcattcag agcctgaaag acgatccctc gcagtctgca aatctcctgg ccgaagccaa
     2161 gaagctgaat gacgcacagg ctccaaaagt tgataacaag tttaacaagg agcagcagaa
     2221 tgctttctac gagatattgc accttcctaa tctgaatgag gaacaaagaa atgcatttat
     2281 ccagtcccta aaagacgatc catcacagag tgccaacctg cttgcggaag ccaaaaagct
     2341 caatgatgcc caggccccga aagtagacgc gaaccatcaa taaggatccg caggcctctg
     2401 ctagcttgac tgactgagat acagcgtacc ttcagctcac agacatgata agatacattg
     2461 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     2521 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     2581 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     2641 aaaacctcta caaatgtggt attggcccat ctctatcggt atcgtagcat aaccccttgg
     2701 ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg ggccggattg ctatctaccg
     2761 gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc
     2821 cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac
     2881 cagaaattta tccttaaggt cgtcagctat cctgcaggcg atctctcgat ttcgatcaag
     2941 acattccttt aatggtcttt tctggacacc actaggggtc agaagtagtt catcaaactt
     3001 tcttccctcc ctaatctcat tggttacctt gggctatcga aacttaatta accagtcaag
     3061 tcagctactt ggcgagatcg acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc
     3121 aagttcgatc tggtccttgc tattgcaccc gttctccgat tacgagtttc atttaaatca
     3181 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3241 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3301 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3361 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     3421 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     3481 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     3541 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3601 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3661 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3721 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3781 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3841 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3901 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3961 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4021 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcatttaaa tttccgaact
     4081 ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt gcaggctacc
     4141 tctcgaacga actatcgcaa gtctcttggc cggccttgcg ccttggctat tgcttggcag
     4201 cgcctatcgc caggtattac tccaatcccg aatatccgag atcgggatca cccgagagaa
     4261 gttcaaccta catcctcaat cccgatctat ccgagatccg aggaatatcg aaatcggggc
     4321 gcgcctggcc tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc gtcacggcga
     4381 gcgctgccac gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc ggacgctcag
     4441 gacagcggcc cgctgctcat aagactcggc cttagaaccc cagtatcagc agaaggacat
     4501 tttaggacgg gacttgggtg actctagggc actggttttc tttccagaga gcggaacagg
     4561 cgaggaaaag tagtcccttc tcggcgattc tgcggaggga tctccgtggg gcggtgaacg
     4621 ccgatgatta tataaggacg cgccgggtgt ggcacagcta gttccgtcgc agccgggatt
     4681 tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta gcgggctgct
     4741 gggctggccg gggctttcgt ggccgccggg ccgctcggtg ggacggaagc gtgtggagag
     4801 accgccaagg gctgtagtct gggtccgcga gcaaggttgc cctgaactgg gggttggggg
     4861 gagcgcagca aaatggcggc tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc
     4921 tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg caagaaccca aggtcttgag
     4981 cccttcgcta atgcgggaaa gctcttattc gggtgagatg ggctgggcac catctgggga
     5041 ccctgacgtg aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg
     5101 cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg
     5161 tgtcgtgacg tcacccgttc tgttggctta taatgcaggg tggggccacc tgccggtagg
     5221 tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct
     5281 gaatcgacag gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg
     5341 gtcggtttta tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc
     5401 ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg
     5461 gtcaatatgt aattttcagt gttagacttg taaattgtcc gctaaattct ggccgttttt
     5521 ggcttttttg ttagacaaca tgggtaaaaa gcctgaactc accgcgacgt ctgtcgagaa
     5581 gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctcgg agggcgaaga
     5641 atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg taaatagctg
     5701 cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatccg ccgcgctccc
     5761 gattccggaa gtgcttgaca ttggggagtt cagcgagagc ctgacctatt gcatctcccg
     5821 ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg ctgttctgca
     5881 gccggtcgcg gaggcaatgg atgccatcgc tgccgccgat cttagccaga cgagcgggtt
     5941 cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt tcatatgcgc
     6001 gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg tcagtgcgtc
     6061 cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg aagtccggca
     6121 cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc gcataacagc
     6181 ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg ccaacatctt
     6241 cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg agcggaggca
     6301 tccggagctt gcaggatcgc cccggctccg ggcgtatatg ctccgcattg gtcttgacca
     6361 actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc agggtcgatg
     6421 cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg cccgcagaag
     6481 cgccgccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa accgacgccc
     6541 cagcactcgt ccgagggcaa aggaataagc tagtatgtaa gcctagtctt agataataaa
     6601 atcgctatcc atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg ctaggcgcgc
     6661 ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc
     6721 agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc
     6781 aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt
     6841 caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag
     6901 gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     6961 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     7021 tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     7081 ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     7141 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     7201 atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     7261 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     7321 caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     7381 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     7441 ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta
     7501 ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac
     7561 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     7621 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     7681 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     7741 taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa
     7801 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     7861 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga
     7921 tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta cgcgaacagc
     7981 cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag cttacctttt
     8041 tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat cttcttccct
     8101 ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct caagacccgt
     8161 ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa aaaccaacac
     8221 acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt gtagccagat
     8281 ctagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     8341 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     8401 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     8461 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     8521 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     8581 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatgctgat
     8641 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     8701 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     8761 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     8821 ggtctatata agcagagctg gtttagtgaa ccgtcagatc agatctttgt cgatcctacc
     8881 atccactcga cacacccgcc agcggccgc
//
