LOCUS       sequence-295853-        7942 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 7942)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7942)
  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-m4133858
FEATURES             Location/Qualifiers
     source          1..7942
                     /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..1858
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1911..2032
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1922..1941)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1976..1995
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2147..2193
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2223..2359
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2404..2475
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2701..3289)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2781..2800)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3789..4998
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4835..4854
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             5002..5601
                     /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(5002..5021)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5498..5518
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5628..5676
                     /note="synthetic polyadenylation signal"
     CDS             join(5942..6010,6011..6802)
                     /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(6160..6179)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6815..6951
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7096..7167
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7172..7218)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7236..7284
                     /note="synthetic polyadenylation signal"
     enhancer        7365..7668
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7670..7872
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7823..7843
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(7941..7942,1..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 gtacccctat gacgtaccag attatgcgta
     1861 agctagcttg actgactgag atacagcgta ccttcagctc acagacatga taagatacat
     1921 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     1981 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     2041 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     2101 gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc ataacccctt
     2161 ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat tgctatctac
     2221 cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
     2281 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
     2341 accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg atttcgatca
     2401 agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag ttcatcaaac
     2461 tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat taaccagtca
     2521 agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga attgcgtcag
     2581 tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt tcatttaaat
     2641 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2701 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2761 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2821 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2881 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2941 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3001 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3061 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3121 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3181 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3241 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3301 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3361 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3421 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3481 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta aatttccgaa
     3541 ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc ttgcaggcta
     3601 cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct attgcttggc
     3661 agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat cacccgagag
     3721 aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat cgaaatcggg
     3781 gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc tcgtcacggc
     3841 gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg atccttccgc ccggacgctc
     3901 aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca gcagaaggac
     3961 attttaggac gggacttggg tgactctagg gcactggttt tctttccaga gagcggaaca
     4021 ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg gggcggtgaa
     4081 cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc gcagccggga
     4141 tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag tagcgggctg
     4201 ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa gcgtgtggag
     4261 agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact gggggttggg
     4321 gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct tgtgaggcgg
     4381 gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc caaggtcttg
     4441 agcccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggc accatctggg
     4501 gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg ttgcgggggc
     4561 ggcagttatg gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc gcgccctcgt
     4621 cgtgtcgtga cgtcacccgt tctgttggct tataatgcag ggtggggcca cctgccggta
     4681 ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt cgggcctagg gtaggctctc
     4741 ctgaatcgac aggcgccgga cctctggtga ggggagggat aagtgaggcg tcagtttctt
     4801 tggtcggttt tatgtaccta tcttcttaag tagctgaagc tccggttttg aactatgcgc
     4861 tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat gtaatcattt
     4921 gggtcaatat gtaattttca gtgttagact tgtaaattgt ccgctaaatt ctggccgttt
     4981 ttggcttttt tgttagacaa catgaccgag tacaagccca cggtgcgcct cgccacccgc
     5041 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     5101 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     5161 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     5221 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     5281 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga gggcctcctg
     5341 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     5401 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc cgccgagcgc
     5461 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     5521 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     5581 acccgcaagc ccggtgccta ggctagtatg taagcctagt cttagataat aaaatcgcta
     5641 tccatcgaag atggatgtgt gttggttttt tgtgtgtgta acgctaggcg cgcctggtgt
     5701 accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt ggcagtcagc
     5761 cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta ctcaagcctg
     5821 gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc ggtcaacgta
     5881 taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag gaggctttcg
     5941 catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     6001 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     6061 gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     6121 cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     6181 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     6241 ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     6301 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     6361 tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     6421 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     6481 gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac ttactctagc
     6541 ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     6601 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     6661 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     6721 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     6781 ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa aaatgcctga
     6841 tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc
     6901 ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta agatcgttta
     6961 aactcgactc tggctctatc gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg
     7021 ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt cagcttacct ttttggcagc
     7081 gatcgcggct cccgacatct tggaccatta gctccacagg tatcttcttc cctctagtgg
     7141 tcataacagc agcttcagct acctctcaat tcaaaaaacc cctcaagacc cgtttagagg
     7201 ccccaagggg ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa cacacatcca
     7261 tcttcgatgg atagcgattt tattatctaa ctgctgatcg agtgtagcca gatctagtaa
     7321 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     7381 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     7441 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     7501 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     7561 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     7621 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatgct gatgcggttt
     7681 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     7741 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     7801 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     7861 ataagcagag ctggtttagt gaaccgtcag atcagatctt tgtcgatcct accatccact
     7921 cgacacaccc gccagcggcc gc
//
