LOCUS       sequence-295581-        6399 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 6399)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6399)
  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-m4133787
FEATURES             Location/Qualifiers
     source          1..6399
                     /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             2174..2191
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2268..2389
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2279..2298)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2333..2352
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2504..2550
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2580..2716
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2761..2832
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3058..3646)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3138..3157)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4392..4460,4461..5252)
                     /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(4610..4629)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5265..5401
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5553..5624
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5629..5675)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5693..5741
                     /note="synthetic polyadenylation signal"
     enhancer        5822..6125
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6127..6329
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6280..6300
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6398..6399,1..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 agtcgatgag tgacaaaatc attcatctaa ccgatgactc
     1861 ctttgataca gacgtgttga aggccgatgg tgcgatactg gtagatttct gggctgaatg
     1921 gtgtggcccg tgcaagatga tcgcacccat tctggacgag atcgccgacg agtatcaagg
     1981 aaaattaact gtcgctaagc ttaatataga tcagaaccca gggacggcac ctaaatacgg
     2041 catcagggga atacctaccc tcctgctctt taagaatggg gaagttgccg cgacaaaagt
     2101 gggagccctg tctaagggcc agctaaaaga gttcttggac gctaacctgg caggttcagg
     2161 ctcggggcac atgcaccatc accatcacca tagcagcggg ctcgtgccac gcggctaagc
     2221 tagcttgact gactgagata cagcgtacct tcagctcaca gacatgataa gatacattga
     2281 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     2341 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     2401 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     2461 aaacctctac aaatgtggta ttggcccatc tctatcggta tcgtagcata accccttggg
     2521 gcctctaaac gggtcttgag gggttttttg tgcccctcgg gccggattgc tatctaccgg
     2581 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
     2641 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
     2701 agaaatttat ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga
     2761 cattccttta atggtctttt ctggacacca ctaggggtca gaagtagttc atcaaacttt
     2821 cttccctccc taatctcatt ggttaccttg ggctatcgaa acttaattaa ccagtcaagt
     2881 cagctacttg gcgagatcga cttgtctggg tttcgactac gctcagaatt gcgtcagtca
     2941 agttcgatct ggtccttgct attgcacccg ttctccgatt acgagtttca tttaaatcat
     3001 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3061 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3121 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3181 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3241 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3301 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3361 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3421 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3481 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3541 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3601 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3661 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3721 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3781 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     3841 acctatctca gcgatctgtc tatttcgttc atccatagtt gcatttaaat ttccgaactc
     3901 tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg caggctacct
     3961 ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc cttggctatt gcttggcagc
     4021 gcctatcgcc aggtattact ccaatcccga atatccgaga tcgggatcac ccgagagaag
     4081 ttcaacctac atcctcaatc ccgatctatc cgagatccga ggaatatcga aatcggggcg
     4141 cgcctggtgt accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt
     4201 ggcagtcagc cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta
     4261 ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc
     4321 ggtcaacgta taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag
     4381 gaggctttcg catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     4441 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     4501 agttgggtgc gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     4561 gttttcgccc cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4621 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4681 agaatgactt ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4741 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4801 tgacaacgat tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4861 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4921 acaccacgat gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac
     4981 ttactctagc ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac
     5041 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5101 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5161 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5221 agataggtgc ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa
     5281 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     5341 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     5401 agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta
     5461 cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag
     5521 cttacctttt tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat
     5581 cttcttccct ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct
     5641 caagacccgt ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa
     5701 aaaccaacac acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt
     5761 gtagccagat ctagtaatca attacggggt cattagttca tagcccatat atggagttcc
     5821 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     5881 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     5941 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     6001 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     6061 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     6121 ccatgctgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     6181 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     6241 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     6301 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc agatctttgt
     6361 cgatcctacc atccactcga cacacccgcc agcggccgc
//
