LOCUS       sequence-295427-        8305 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 8305)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 8305)
  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-m4133394
FEATURES             Location/Qualifiers
     source          1..8305
                     /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    2274..2395
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2285..2304)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2339..2358
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2510..2556
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2586..2722
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2767..2838
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3064..3652)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3144..3163)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4152..5361
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     5198..5217
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5365..5964
                     /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(5365..5384)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5861..5881
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5991..6039
                     /note="synthetic polyadenylation signal"
     CDS             join(6305..6373,6374..7165)
                     /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(6523..6542)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7178..7314
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7459..7530
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7535..7581)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7599..7647
                     /note="synthetic polyadenylation signal"
     enhancer        7728..8031
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8033..8235
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8186..8206
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(8304..8305,1..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 ctaagctagc ttgactgact gagatacagc gtaccttcag ctcacagaca tgataagata
     2281 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     2341 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     2401 caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag
     2461 caagtaaaac ctctacaaat gtggtattgg cccatctcta tcggtatcgt agcataaccc
     2521 cttggggcct ctaaacgggt cttgaggggt tttttgtgcc cctcgggccg gattgctatc
     2581 taccggcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca
     2641 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc
     2701 cttaccagaa atttatcctt aaggtcgtca gctatcctgc aggcgatctc tcgatttcga
     2761 tcaagacatt cctttaatgg tcttttctgg acaccactag gggtcagaag tagttcatca
     2821 aactttcttc cctccctaat ctcattggtt accttgggct atcgaaactt aattaaccag
     2881 tcaagtcagc tacttggcga gatcgacttg tctgggtttc gactacgctc agaattgcgt
     2941 cagtcaagtt cgatctggtc cttgctattg cacccgttct ccgattacga gtttcattta
     3001 aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3061 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3121 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3181 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3241 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3301 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3361 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3421 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3481 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3541 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3601 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3661 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3721 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3781 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3841 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcat ttaaatttcc
     3901 gaactctcca aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc atcttgcagg
     3961 ctacctctcg aacgaactat cgcaagtctc ttggccggcc ttgcgccttg gctattgctt
     4021 ggcagcgcct atcgccaggt attactccaa tcccgaatat ccgagatcgg gatcacccga
     4081 gagaagttca acctacatcc tcaatcccga tctatccgag atccgaggaa tatcgaaatc
     4141 ggggcgcgcc tggcctccgc gccgggtttt ggcgcctccc gcgggcgccc ccctcgtcac
     4201 ggcgagcgct gccacgtcag acgaagggcg caggagcgtc ctgatccttc cgcccggacg
     4261 ctcaggacag cggcccgctg ctcataagac tcggccttag aaccccagta tcagcagaag
     4321 gacattttag gacgggactt gggtgactct agggcactgg ttttctttcc agagagcgga
     4381 acaggcgagg aaaagtagtc ccttctcggc gattctgcgg agggatctcc gtggggcggt
     4441 gaacgccgat gattatataa ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg
     4501 ggatttgggt cgcggttctt gtttgtggat cgctgtgatc gtcacttggt gagtagcggg
     4561 ctgctgggct ggccggggct ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg
     4621 gagagaccgc caagggctgt agtctgggtc cgcgagcaag gttgccctga actgggggtt
     4681 ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg aatggaagac gcttgtgagg
     4741 cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga acccaaggtc
     4801 ttgagccctt cgctaatgcg ggaaagctct tattcgggtg agatgggctg ggcaccatct
     4861 ggggaccctg acgtgaagtt tgtcactgac tggagaactc ggtttgtcgt ctgttgcggg
     4921 ggcggcagtt atggcggtgc cgttgggcag tgcacccgta cctttgggag cgcgcgccct
     4981 cgtcgtgtcg tgacgtcacc cgttctgttg gcttataatg cagggtgggg ccacctgccg
     5041 gtaggtgtgc ggtaggcttt tctccgtcgc aggacgcagg gttcgggcct agggtaggct
     5101 ctcctgaatc gacaggcgcc ggacctctgg tgaggggagg gataagtgag gcgtcagttt
     5161 ctttggtcgg ttttatgtac ctatcttctt aagtagctga agctccggtt ttgaactatg
     5221 cgctcggggt tggcgagtgt gttttgtgaa gttttttagg caccttttga aatgtaatca
     5281 tttgggtcaa tatgtaattt tcagtgttag acttgtaaat tgtccgctaa attctggccg
     5341 tttttggctt ttttgttaga caacatgacc gagtacaagc ccacggtgcg cctcgccacc
     5401 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     5461 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     5521 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     5581 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     5641 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggagggcctc
     5701 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
     5761 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggccgccgag
     5821 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
     5881 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     5941 atgacccgca agcccggtgc ctaggctagt atgtaagcct agtcttagat aataaaatcg
     6001 ctatccatcg aagatggatg tgtgttggtt ttttgtgtgt gtaacgctag gcgcgcctgg
     6061 tgtaccgaga acgatcctct cagtgcgagt ctcgacgatc catatcgttg cttggcagtc
     6121 agccagtcgg aatccagctt gggacccagg aagtccaatc gtcagatatt gtactcaagc
     6181 ctggtcacgg cagcgtaccg atctgtttaa acctagatat tgatagtctg atcggtcaac
     6241 gtataatcga gtcctagctt ttgcaaacat ctatcaagag acaggatcag caggaggctt
     6301 tcgcatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     6361 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     6421 tgcgcgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     6481 ccccgaagaa cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     6541 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     6601 cttggttgag tattcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     6661 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     6721 gattggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     6781 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     6841 gatgcctgta gcaatggcaa caaccttgcg taaactatta actggcgaac tacttactct
     6901 agcttcccgg caacagttga tagactggat ggaggcggat aaagttgcag gaccacttct
     6961 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     7021 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     7081 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     7141 tgcctcactg attaagcatt ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc
     7201 tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct
     7261 tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaagatcgt
     7321 ttaaactcga ctctggctct atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg
     7381 gcgctcattt gctcgtcggg catcgaatct cgtcagctat cgtcagctta cctttttggc
     7441 agcgatcgcg gctcccgaca tcttggacca ttagctccac aggtatcttc ttccctctag
     7501 tggtcataac agcagcttca gctacctctc aattcaaaaa acccctcaag acccgtttag
     7561 aggccccaag gggttatgct atcaatcgtt gcgttacaca cacaaaaaac caacacacat
     7621 ccatcttcga tggatagcga ttttattatc taactgctga tcgagtgtag ccagatctag
     7681 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     7741 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     7801 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     7861 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     7921 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     7981 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat gctgatgcgg
     8041 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     8101 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     8161 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     8221 tatataagca gagctggttt agtgaaccgt cagatcagat ctttgtcgat cctaccatcc
     8281 actcgacaca cccgccagcg gccgc
//
