LOCUS       sequence-295033-        8405 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 8405)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8405)
  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-m4133416
FEATURES             Location/Qualifiers
     source          1..8405
                     /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..1876
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    1945..2066
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1956..1975)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2010..2029
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2181..2227
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2257..2393
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2438..2509
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2735..3323)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2815..2834)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3823..5032
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4869..4888
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5036..6064
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6091..6139
                     /note="synthetic polyadenylation signal"
     CDS             join(6405..6473,6474..7265)
                     /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(6623..6642)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7278..7414
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7559..7630
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7635..7681)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7699..7747
                     /note="synthetic polyadenylation signal"
     enhancer        7828..8131
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8133..8335
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8286..8306
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(8404..8405,1..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 gaaagaaacc gcggcagcta agttcgagag
     1861 gcagcacatg gacagttaag gatccgcagg cctctgctag cttgactgac tgagatacag
     1921 cgtaccttca gctcacagac atgataagat acattgatga gtttggacaa accacaacta
     1981 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     2041 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     2101 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg
     2161 gcccatctct atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg
     2221 ttttttgtgc ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct
     2281 gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt
     2341 ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc
     2401 agctatcctg caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg
     2461 gacaccacta ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt
     2521 taccttgggc tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt
     2581 gtctgggttt cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt
     2641 gcacccgttc tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg
     2701 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2761 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2821 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2881 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2941 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3001 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3061 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3121 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3181 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3241 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3301 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3361 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     3421 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     3481 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3541 ttcgttcatc catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc
     3601 ttcaaacctt tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct
     3661 cttggccggc cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca
     3721 atcccgaata tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg
     3781 atctatccga gatccgagga atatcgaaat cggggcgcgc ctggcctccg cgccgggttt
     3841 tggcgcctcc cgcgggcgcc cccctcgtca cggcgagcgc tgccacgtca gacgaagggc
     3901 gcaggagcgt cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga
     3961 ctcggcctta gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc
     4021 tagggcactg gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg
     4081 cgattctgcg gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc
     4141 gggtgtggca cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga
     4201 tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc
     4261 gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt
     4321 ccgcgagcaa ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt
     4381 cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg
     4441 ggggcatggt gggcggcaag aacccaaggt cttgagccct tcgctaatgc gggaaagctc
     4501 ttattcgggt gagatgggct gggcaccatc tggggaccct gacgtgaagt ttgtcactga
     4561 ctggagaact cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca
     4621 gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt
     4681 ggcttataat gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg
     4741 caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg
     4801 gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct
     4861 taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga
     4921 agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta
     4981 gacttgtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag acaacatggg
     5041 taaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag
     5101 cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt
     5161 aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg
     5221 ttatgtttat cggcactttg catccgccgc gctcccgatt ccggaagtgc ttgacattgg
     5281 ggagttcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca
     5341 agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg caatggatgc
     5401 catcgctgcc gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat
     5461 cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca
     5521 ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct
     5581 gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc
     5641 caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat
     5701 gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg
     5761 tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccccg
     5821 gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg
     5881 caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc
     5941 cgggactgtc gggcgtacac aaatcgcccg cagaagcgcc gccgtctgga ccgatggctg
     6001 tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga
     6061 ataagctagt atgtaagcct agtcttagat aataaaatcg ctatccatcg aagatggatg
     6121 tgtgttggtt ttttgtgtgt gtaacgctag gcgcgcctgg tgtaccgaga acgatcctct
     6181 cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt
     6241 gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg
     6301 atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt
     6361 ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt
     6421 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     6481 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg
     6541 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa
     6601 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     6661 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag
     6721 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     6781 ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc
     6841 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     6901 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     6961 caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg caacagttga
     7021 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     7081 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     7141 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     7201 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     7261 ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
     7321 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
     7381 atacgtgtcc tccttaccag aaatttatcc ttaagatcgt ttaaactcga ctctggctct
     7441 atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt gctcgtcggg
     7501 catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg gctcccgaca
     7561 tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac agcagcttca
     7621 gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct
     7681 atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga tggatagcga
     7741 ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta cggggtcatt
     7801 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     7861 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     7921 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     7981 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     8041 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     8101 catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt acatcaatgg
     8161 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     8221 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     8281 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
     8341 agtgaaccgt cagatcagat ctttgtcgat cctaccatcc actcgacaca cccgccagcg
     8401 gccgc
//
