LOCUS       sequence-295719-        7987 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 7987)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7987)
  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-m4133921
FEATURES             Location/Qualifiers
     source          1..7987
                     /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             1826..1903
                     /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"
     polyA_signal    1956..2077
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1967..1986)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2021..2040
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2192..2238
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2268..2404
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2449..2520
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2746..3334)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2826..2845)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3834..5043
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4880..4899
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5047..5646
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5047..5066)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5543..5563
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5673..5721
                     /note="synthetic polyadenylation signal"
     CDS             join(5987..6055,6056..6847)
                     /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(6205..6224)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6860..6996
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7141..7212
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7217..7263)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7281..7329
                     /note="synthetic polyadenylation signal"
     enhancer        7410..7713
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7715..7917
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7868..7888
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(7986..7987,1..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 agtcgaagag gcgatggaaa aagaatttta tcgcggtgag
     1861 tgcagccaac agattcaaaa agatatcctc gtctggtgcc ctctaagcta gcttgactga
     1921 ctgagataca gcgtaccttc agctcacaga catgataaga tacattgatg agtttggaca
     1981 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     2041 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     2101 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     2161 atgtggtatt ggcccatctc tatcggtatc gtagcataac cccttggggc ctctaaacgg
     2221 gtcttgaggg gttttttgtg cccctcgggc cggattgcta tctaccggca ttggcgcaga
     2281 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
     2341 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
     2401 ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc gatcaagaca ttcctttaat
     2461 ggtcttttct ggacaccact aggggtcaga agtagttcat caaactttct tccctcccta
     2521 atctcattgg ttaccttggg ctatcgaaac ttaattaacc agtcaagtca gctacttggc
     2581 gagatcgact tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg
     2641 tccttgctat tgcacccgtt ctccgattac gagtttcatt taaatcatgt gagcaaaagg
     2701 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2761 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2821 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2881 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2941 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3001 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3061 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3121 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3181 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3241 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3301 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3361 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3421 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3481 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3541 gatctgtcta tttcgttcat ccatagttgc atttaaattt ccgaactctc caaggccctc
     3601 gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca ggctacctct cgaacgaact
     3661 atcgcaagtc tcttggccgg ccttgcgcct tggctattgc ttggcagcgc ctatcgccag
     3721 gtattactcc aatcccgaat atccgagatc gggatcaccc gagagaagtt caacctacat
     3781 cctcaatccc gatctatccg agatccgagg aatatcgaaa tcggggcgcg cctggcctcc
     3841 gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcgtc acggcgagcg ctgccacgtc
     3901 agacgaaggg cgcaggagcg tcctgatcct tccgcccgga cgctcaggac agcggcccgc
     3961 tgctcataag actcggcctt agaaccccag tatcagcaga aggacatttt aggacgggac
     4021 ttgggtgact ctagggcact ggttttcttt ccagagagcg gaacaggcga ggaaaagtag
     4081 tcccttctcg gcgattctgc ggagggatct ccgtggggcg gtgaacgccg atgattatat
     4141 aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc
     4201 ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg ctggccgggg
     4261 ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg tggagagacc gccaagggct
     4321 gtagtctggg tccgcgagca aggttgccct gaactggggg ttggggggag cgcagcaaaa
     4381 tggcggctgt tcccgagtct tgaatggaag acgcttgtga ggcgggctgt gaggtcgttg
     4441 aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg tcttgagccc ttcgctaatg
     4501 cgggaaagct cttattcggg tgagatgggc tgggcaccat ctggggaccc tgacgtgaag
     4561 tttgtcactg actggagaac tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt
     4621 gccgttgggc agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca
     4681 cccgttctgt tggcttataa tgcagggtgg ggccacctgc cggtaggtgt gcggtaggct
     4741 tttctccgtc gcaggacgca gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg
     4801 ccggacctct ggtgagggga gggataagtg aggcgtcagt ttctttggtc ggttttatgt
     4861 acctatcttc ttaagtagct gaagctccgg ttttgaacta tgcgctcggg gttggcgagt
     4921 gtgttttgtg aagtttttta ggcacctttt gaaatgtaat catttgggtc aatatgtaat
     4981 tttcagtgtt agacttgtaa attgtccgct aaattctggc cgtttttggc ttttttgtta
     5041 gacaacatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
     5101 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
     5161 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     5221 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     5281 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     5341 agcggttccc ggctggccgc gcagcaacag atggagggcc tcctggcgcc gcaccggccc
     5401 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     5461 ctgggcagcg ccgtcgtgct ccccggagtg gaggccgccg agcgcgccgg ggtgcccgcc
     5521 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     5581 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
     5641 gcctaggcta gtatgtaagc ctagtcttag ataataaaat cgctatccat cgaagatgga
     5701 tgtgtgttgg ttttttgtgt gtgtaacgct aggcgcgcct ggtgtaccga gaacgatcct
     5761 ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc ggaatccagc
     5821 ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac
     5881 cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc gagtcctagc
     5941 ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga gtattcaaca
     6001 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     6061 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag tgggttacat
     6121 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgctttcc
     6181 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     6241 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtattcacc
     6301 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     6361 aaccatgagt gataacactg cggccaactt acttctgaca acgattggag gaccgaagga
     6421 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     6481 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     6541 aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc ggcaacagtt
     6601 gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     6661 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     6721 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     6781 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     6841 ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt
     6901 cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt
     6961 ccatacgtgt cctccttacc agaaatttat ccttaagatc gtttaaactc gactctggct
     7021 ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg
     7081 ggcatcgaat ctcgtcagct atcgtcagct tacctttttg gcagcgatcg cggctcccga
     7141 catcttggac cattagctcc acaggtatct tcttccctct agtggtcata acagcagctt
     7201 cagctacctc tcaattcaaa aaacccctca agacccgttt agaggcccca aggggttatg
     7261 ctatcaatcg ttgcgttaca cacacaaaaa accaacacac atccatcttc gatggatagc
     7321 gattttatta tctaactgct gatcgagtgt agccagatct agtaatcaat tacggggtca
     7381 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     7441 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     7501 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     7561 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     7621 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     7681 tacatctacg tattagtcat cgctattacc atgctgatgc ggttttggca gtacatcaat
     7741 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     7801 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     7861 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt
     7921 ttagtgaacc gtcagatcag atctttgtcg atcctaccat ccactcgaca cacccgccag
     7981 cggccgc
//
