LOCUS       sequence-295783-        6057 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 6057)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6057)
  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-m4133260
FEATURES             Location/Qualifiers
     source          1..6057
                     /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..1873
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    1926..2047
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1937..1956)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1991..2010
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2162..2208
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2238..2374
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2419..2490
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2716..3304)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2796..2815)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4050..4118,4119..4910)
                     /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(4268..4287)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4923..5059
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5211..5282
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5287..5333)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5351..5399
                     /note="synthetic polyadenylation signal"
     enhancer        5480..5783
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5785..5987
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5938..5958
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6056..6057,1..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 ggggaaacct atacccaatc cacttctcgg
     1861 cttagattct acctaagcta gcttgactga ctgagataca gcgtaccttc agctcacaga
     1921 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     1981 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     2041 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     2101 ggttttttaa agcaagtaaa acctctacaa atgtggtatt ggcccatctc tatcggtatc
     2161 gtagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc
     2221 cggattgcta tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
     2281 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
     2341 atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc
     2401 tctcgatttc gatcaagaca ttcctttaat ggtcttttct ggacaccact aggggtcaga
     2461 agtagttcat caaactttct tccctcccta atctcattgg ttaccttggg ctatcgaaac
     2521 ttaattaacc agtcaagtca gctacttggc gagatcgact tgtctgggtt tcgactacgc
     2581 tcagaattgc gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt ctccgattac
     2641 gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2701 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2761 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2821 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2881 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2941 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3001 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3061 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3121 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     3181 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3241 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3301 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3361 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3421 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3481 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3541 atttaaattt ccgaactctc caaggccctc gtcggaaaat cttcaaacct ttcgtccgat
     3601 ccatcttgca ggctacctct cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct
     3661 tggctattgc ttggcagcgc ctatcgccag gtattactcc aatcccgaat atccgagatc
     3721 gggatcaccc gagagaagtt caacctacat cctcaatccc gatctatccg agatccgagg
     3781 aatatcgaaa tcggggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga
     3841 cgatccatat cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc
     3901 caatcgtcag atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta
     3961 gatattgata gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc
     4021 aagagacagg atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta
     4081 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     4141 taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca
     4201 gcggtaagat ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta
     4261 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     4321 gccgcataca ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc
     4381 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     4441 ctgcggccaa cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc
     4501 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     4561 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac
     4621 tattaactgg cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg
     4681 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     4741 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     4801 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     4861 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag
     4921 gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
     4981 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
     5041 accagaaatt tatccttaag atcccgaatc gtttaaactc gactctggct ctatcgaatc
     5101 tccgtcgttt cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat
     5161 ctcgtcagct atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac
     5221 cattagctcc acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc
     5281 tcaattcaaa aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg
     5341 ttgcgttaca cacacaaaaa accaacacac atccatcttc gatggatagc gattttatta
     5401 tctaactgct gatcgagtgt agccagatct agtaatcaat tacggggtca ttagttcata
     5461 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     5521 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     5581 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     5641 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     5701 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     5761 tattagtcat cgctattacc atgctgatgc ggttttggca gtacatcaat gggcgtggat
     5821 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     5881 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     5941 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
     6001 gtcagatcag atctttgtcg atcctaccat ccactcgaca cacccgccag cggccgc
//
