LOCUS       sequence-295056-        8915 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 8915)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8915)
  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-m4134365
FEATURES             Location/Qualifiers
     source          1..8915
                     /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             1865..1942
                     /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             1970..1990
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    2455..2576
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2466..2485)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2520..2539
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2691..2737
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2767..2903
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2948..3019
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3245..3833)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3325..3344)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4333..5542
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5379..5398
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5546..6574
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    6601..6649
                     /note="synthetic polyadenylation signal"
     CDS             join(6915..6983,6984..7775)
                     /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(7133..7152)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7788..7924
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    8069..8140
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(8145..8191)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    8209..8257
                     /note="synthetic polyadenylation signal"
     enhancer        8338..8641
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8643..8845
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8796..8816
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(8914..8915,1..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 gggacggcga atcccggggc tgataaatcc
     1861 ctggaaaagg agatggaaga agaactttat tgcagtgtct gcggctaacc ggttcaaaaa
     1921 gatcagttca agcggcgccc tagattacga cattcctacg accgcctccg agaatctcta
     1981 tttccagggt gaatttggct tggcacaaca tgacgaggcc gtcgataaca agtttaataa
     2041 agaacagcag aacgctttct atgagatcct tcacttacca aatctgaacg aagagcaacg
     2101 caacgctttc attcagagcc tgaaagacga tccctcgcag tctgcaaatc tcctggccga
     2161 agccaagaag ctgaatgacg cacaggctcc aaaagttgat aacaagttta acaaggagca
     2221 gcagaatgct ttctacgaga tattgcacct tcctaatctg aatgaggaac aaagaaatgc
     2281 atttatccag tccctaaaag acgatccatc acagagtgcc aacctgcttg cggaagccaa
     2341 aaagctcaat gatgcccagg ccccgaaagt agacgcgaac catcaataag gatccgcagg
     2401 cctctgctag cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat
     2461 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     2521 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     2581 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
     2641 gcaagtaaaa cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc
     2701 ccttggggcc tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat
     2761 ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac
     2821 atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct
     2881 ccttaccaga aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg
     2941 atcaagacat tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc
     3001 aaactttctt ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca
     3061 gtcaagtcag ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg
     3121 tcagtcaagt tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt
     3181 aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3241 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3301 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3361 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3421 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3481 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3541 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     3601 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     3661 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     3721 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3781 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3841 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3901 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3961 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4021 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc
     4081 cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag
     4141 gctacctctc gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct
     4201 tggcagcgcc tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg
     4261 agagaagttc aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat
     4321 cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc cccctcgtca
     4381 cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt cctgatcctt ccgcccggac
     4441 gctcaggaca gcggcccgct gctcataaga ctcggcctta gaaccccagt atcagcagaa
     4501 ggacatttta ggacgggact tgggtgactc tagggcactg gttttctttc cagagagcgg
     4561 aacaggcgag gaaaagtagt cccttctcgg cgattctgcg gagggatctc cgtggggcgg
     4621 tgaacgccga tgattatata aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc
     4681 gggatttggg tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg
     4741 gctgctgggc tggccggggc tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt
     4801 ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg aactgggggt
     4861 tggggggagc gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag
     4921 gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt gggcggcaag aacccaaggt
     4981 cttgagccct tcgctaatgc gggaaagctc ttattcgggt gagatgggct gggcaccatc
     5041 tggggaccct gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg
     5101 gggcggcagt tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc
     5161 tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccacctgcc
     5221 ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc
     5281 tctcctgaat cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt
     5341 tctttggtcg gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat
     5401 gcgctcgggg ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc
     5461 atttgggtca atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc
     5521 gtttttggct tttttgttag acaacatggg taaaaagcct gaactcaccg cgacgtctgt
     5581 cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg
     5641 cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa
     5701 tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg catccgccgc
     5761 gctcccgatt ccggaagtgc ttgacattgg ggagttcagc gagagcctga cctattgcat
     5821 ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt
     5881 tctgcagccg gtcgcggagg caatggatgc catcgctgcc gccgatctta gccagacgag
     5941 cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat
     6001 atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag
     6061 tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt
     6121 ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat
     6181 aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa
     6241 catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg
     6301 gaggcatccg gagcttgcag gatcgccccg gctccgggcg tatatgctcc gcattggtct
     6361 tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg
     6421 tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg
     6481 cagaagcgcc gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg
     6541 acgccccagc actcgtccga gggcaaagga ataagctagt atgtaagcct agtcttagat
     6601 aataaaatcg ctatccatcg aagatggatg tgtgttggtt ttttgtgtgt gtaacgctag
     6661 gcgcgcctgg tgtaccgaga acgatcctct cagtgcgagt ctcgacgatc catatcgttg
     6721 cttggcagtc agccagtcgg aatccagctt gggacccagg aagtccaatc gtcagatatt
     6781 gtactcaagc ctggtcacgg cagcgtaccg atctgtttaa acctagatat tgatagtctg
     6841 atcggtcaac gtataatcga gtcctagctt ttgcaaacat ctatcaagag acaggatcag
     6901 caggaggctt tcgcatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     6961 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     7021 atcagttggg tgcgcgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     7081 agagttttcg ccccgaagaa cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg
     7141 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     7201 ctcagaatga cttggttgag tattcaccag tcacagaaaa gcatcttacg gatggcatga
     7261 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     7321 ttctgacaac gattggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     7381 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     7441 gtgacaccac gatgcctgta gcaatggcaa caaccttgcg taaactatta actggcgaac
     7501 tacttactct agcttcccgg caacagttga tagactggat ggaggcggat aaagttgcag
     7561 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     7621 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     7681 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     7741 ctgagatagg tgcctcactg attaagcatt ggtaaccgat tctaggtgca ttggcgcaga
     7801 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
     7861 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
     7921 ttaagatcgt ttaaactcga ctctggctct atcgaatctc cgtcgtttcg agcttacgcg
     7981 aacagccgtg gcgctcattt gctcgtcggg catcgaatct cgtcagctat cgtcagctta
     8041 cctttttggc agcgatcgcg gctcccgaca tcttggacca ttagctccac aggtatcttc
     8101 ttccctctag tggtcataac agcagcttca gctacctctc aattcaaaaa acccctcaag
     8161 acccgtttag aggccccaag gggttatgct atcaatcgtt gcgttacaca cacaaaaaac
     8221 caacacacat ccatcttcga tggatagcga ttttattatc taactgctga tcgagtgtag
     8281 ccagatctag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     8341 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     8401 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     8461 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     8521 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     8581 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     8641 gctgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     8701 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     8761 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     8821 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatcagat ctttgtcgat
     8881 cctaccatcc actcgacaca cccgccagcg gccgc
//
