LOCUS       sequence-295444-        7954 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 7954)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7954)
  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-m4134090
FEATURES             Location/Qualifiers
     source          1..7954
                     /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..1870
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    1923..2044
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1934..1953)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1988..2007
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2159..2205
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2235..2371
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2416..2487
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2713..3301)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2793..2812)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3801..5010
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4847..4866
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5014..5613
                     /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(5014..5033)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5510..5530
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5640..5688
                     /note="synthetic polyadenylation signal"
     CDS             join(5954..6022,6023..6814)
                     /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(6172..6191)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6827..6963
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7108..7179
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7184..7230)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7248..7296
                     /note="synthetic polyadenylation signal"
     enhancer        7377..7680
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7682..7884
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7835..7855
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(7953..7954,1..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 agtcgaaaga aaccgcggca gctaagttcg agaggcagca
     1861 catggacagt taagctagct tgactgactg agatacagcg taccttcagc tcacagacat
     1921 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     1981 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     2041 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     2101 tttttaaagc aagtaaaacc tctacaaatg tggtattggc ccatctctat cggtatcgta
     2161 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc ctcgggccgg
     2221 attgctatct accggcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat
     2281 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata
     2341 cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag ctatcctgca ggcgatctct
     2401 cgatttcgat caagacattc ctttaatggt cttttctgga caccactagg ggtcagaagt
     2461 agttcatcaa actttcttcc ctccctaatc tcattggtta ccttgggcta tcgaaactta
     2521 attaaccagt caagtcagct acttggcgag atcgacttgt ctgggtttcg actacgctca
     2581 gaattgcgtc agtcaagttc gatctggtcc ttgctattgc acccgttctc cgattacgag
     2641 tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2701 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2761 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2821 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2881 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2941 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3001 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3061 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3121 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3181 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3241 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3301 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3361 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3421 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3481 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcatt
     3541 taaatttccg aactctccaa ggccctcgtc ggaaaatctt caaacctttc gtccgatcca
     3601 tcttgcaggc tacctctcga acgaactatc gcaagtctct tggccggcct tgcgccttgg
     3661 ctattgcttg gcagcgccta tcgccaggta ttactccaat cccgaatatc cgagatcggg
     3721 atcacccgag agaagttcaa cctacatcct caatcccgat ctatccgaga tccgaggaat
     3781 atcgaaatcg gggcgcgcct ggcctccgcg ccgggttttg gcgcctcccg cgggcgcccc
     3841 cctcgtcacg gcgagcgctg ccacgtcaga cgaagggcgc aggagcgtcc tgatccttcc
     3901 gcccggacgc tcaggacagc ggcccgctgc tcataagact cggccttaga accccagtat
     3961 cagcagaagg acattttagg acgggacttg ggtgactcta gggcactggt tttctttcca
     4021 gagagcggaa caggcgagga aaagtagtcc cttctcggcg attctgcgga gggatctccg
     4081 tggggcggtg aacgccgatg attatataag gacgcgccgg gtgtggcaca gctagttccg
     4141 tcgcagccgg gatttgggtc gcggttcttg tttgtggatc gctgtgatcg tcacttggtg
     4201 agtagcgggc tgctgggctg gccggggctt tcgtggccgc cgggccgctc ggtgggacgg
     4261 aagcgtgtgg agagaccgcc aagggctgta gtctgggtcc gcgagcaagg ttgccctgaa
     4321 ctgggggttg gggggagcgc agcaaaatgg cggctgttcc cgagtcttga atggaagacg
     4381 cttgtgaggc gggctgtgag gtcgttgaaa caaggtgggg ggcatggtgg gcggcaagaa
     4441 cccaaggtct tgagcccttc gctaatgcgg gaaagctctt attcgggtga gatgggctgg
     4501 gcaccatctg gggaccctga cgtgaagttt gtcactgact ggagaactcg gtttgtcgtc
     4561 tgttgcgggg gcggcagtta tggcggtgcc gttgggcagt gcacccgtac ctttgggagc
     4621 gcgcgccctc gtcgtgtcgt gacgtcaccc gttctgttgg cttataatgc agggtggggc
     4681 cacctgccgg taggtgtgcg gtaggctttt ctccgtcgca ggacgcaggg ttcgggccta
     4741 gggtaggctc tcctgaatcg acaggcgccg gacctctggt gaggggaggg ataagtgagg
     4801 cgtcagtttc tttggtcggt tttatgtacc tatcttctta agtagctgaa gctccggttt
     4861 tgaactatgc gctcggggtt ggcgagtgtg ttttgtgaag ttttttaggc accttttgaa
     4921 atgtaatcat ttgggtcaat atgtaatttt cagtgttaga cttgtaaatt gtccgctaaa
     4981 ttctggccgt ttttggcttt tttgttagac aacatgaccg agtacaagcc cacggtgcgc
     5041 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
     5101 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
     5161 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     5221 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     5281 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     5341 gagggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     5401 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     5461 gccgccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     5521 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     5581 acctggtgca tgacccgcaa gcccggtgcc taggctagta tgtaagccta gtcttagata
     5641 ataaaatcgc tatccatcga agatggatgt gtgttggttt tttgtgtgtg taacgctagg
     5701 cgcgcctggt gtaccgagaa cgatcctctc agtgcgagtc tcgacgatcc atatcgttgc
     5761 ttggcagtca gccagtcgga atccagcttg ggacccagga agtccaatcg tcagatattg
     5821 tactcaagcc tggtcacggc agcgtaccga tctgtttaaa cctagatatt gatagtctga
     5881 tcggtcaacg tataatcgag tcctagcttt tgcaaacatc tatcaagaga caggatcagc
     5941 aggaggcttt cgcatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     6001 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     6061 tcagttgggt gcgcgagtgg gttacatcga actggatctc aacagcggta agatccttga
     6121 gagttttcgc cccgaagaac gctttccaat gatgagcact tttaaagttc tgctatgtgg
     6181 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     6241 tcagaatgac ttggttgagt attcaccagt cacagaaaag catcttacgg atggcatgac
     6301 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     6361 tctgacaacg attggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     6421 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     6481 tgacaccacg atgcctgtag caatggcaac aaccttgcgt aaactattaa ctggcgaact
     6541 acttactcta gcttcccggc aacagttgat agactggatg gaggcggata aagttgcagg
     6601 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     6661 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     6721 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     6781 tgagataggt gcctcactga ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa
     6841 aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg
     6901 gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct
     6961 taagatcgtt taaactcgac tctggctcta tcgaatctcc gtcgtttcga gcttacgcga
     7021 acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac
     7081 ctttttggca gcgatcgcgg ctcccgacat cttggaccat tagctccaca ggtatcttct
     7141 tccctctagt ggtcataaca gcagcttcag ctacctctca attcaaaaaa cccctcaaga
     7201 cccgtttaga ggccccaagg ggttatgcta tcaatcgttg cgttacacac acaaaaaacc
     7261 aacacacatc catcttcgat ggatagcgat tttattatct aactgctgat cgagtgtagc
     7321 cagatctagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     7381 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     7441 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     7501 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     7561 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     7621 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     7681 ctgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     7741 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     7801 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     7861 tgggaggtct atataagcag agctggttta gtgaaccgtc agatcagatc tttgtcgatc
     7921 ctaccatcca ctcgacacac ccgccagcgg ccgc
//
