LOCUS       sequence-297951-        6084 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 6084)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6084)
  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-m4133221
FEATURES             Location/Qualifiers
     source          1..6084
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    26..74
                     /note="synthetic polyadenylation signal"
     enhancer        155..458
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        460..662
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     613..633
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      731..740
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2543..2563
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2567..2632
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    2685..2806
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2696..2715)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2750..2769
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2921..2967
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2997..3133
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3178..3249
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3475..4063)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3555..3574)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4809..4877,4878..5669)
                     /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(5027..5046)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5682..5818
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5970..6041
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(join(6046..6084,1..8))
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa cacacatcca tcttcgatgg
       61 atagcgattt tattatctaa ctgctgatcg agtgtagcca gatctagtaa tcaattacgg
      121 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      181 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      241 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      301 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      361 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      421 ggcagtacat ctacgtatta gtcatcgcta ttaccatgct gatgcggttt tggcagtaca
      481 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      541 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      601 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      661 ctggtttagt gaaccgtcag atcagatctt tgtcgatcct accatccact cgacacaccc
      721 gccagcggcc gccaccatgg cggtgggggc ttgcgtacta tgtaacagcc agacgtcatt
      781 gcgctgcggt gcatgtataa gaaggccgtt tctgtgttgc aaatgctgct acgatcatgt
      841 tatttcgact tctcacaagc ttgtgctcag tgtcaatcct tatgtgtgta atgcccccgg
      901 atgcgacgtc acagatgtta cccaactgta cctgggcggc atgtcctatt attgtaagag
      961 ccacaaacca ccaatctcct tccccctgtg cgccaacgga caggtgtttg ggctctacaa
     1021 aaacacatgt gtgggatctg acaatgtcac cgacttcaac gctatcgcca catgcgattg
     1081 gactaatgca ggcgactaca ttttagccaa cacctgtacc gaacggctga agttatttgc
     1141 ggctgagact ctaaaggcaa ctgaggaaac cttcaaactt tcatatggta tcgccacggt
     1201 acgtgaggtg ttgtctgatc gagaactgca tctcagttgg gaggtcggga agcctagacc
     1261 tccccttaat cgaaattacg tgttcacagg ttatagggtt accaagaaca gcaaagtcca
     1321 gatcggcgaa tatacttttg agaagggaga ttacggtgac gccgtagtgt atcgtggcac
     1381 aacaacatac aaactgaatg ttggggatta cttcgtgctg acgtcccaca ccgttatgcc
     1441 gctctcagct ccaactttgg ttccccagga acattacgtg cggattactg gactctatcc
     1501 aacgctaaac ataagcgacg agttttccag caacgtggct aattaccaaa aggtcggcat
     1561 gcagaagtac tccacccttc agggtcctcc tgggactggc aaaagtcact tcgccatcgg
     1621 actggcactt tactatccca gtgcgcgcat tgtatatacc gcctgctcac atgcggcagt
     1681 ggacgccctg tgtgagaagg ctttaaagta tctgccaata gacaaatgct ctcgcattat
     1741 ccctgccagg gccagagtgg agtgctttga taagtttaaa gtgaactcga ccctcgaaca
     1801 gtatgtcttc tgcacagtta acgcattgcc cgaaacgacc gccgatatag ttgtgttcga
     1861 cgaaatctca atggctacca attatgatct ctccgtcgtc aatgcacggc tgcgggccaa
     1921 acactacgtc tacatcgggg accccgcgca gttaccagca ccgcgaaccc tactgactaa
     1981 gggcacactt gagccagagt acttcaacag cgtatgtaga ctgatgaaaa ccattggccc
     2041 tgatatgttc ctgggaacat gtcggaggtg ccccgctgaa atagtggaca ccgtttcagc
     2101 acttgtgtac gacaataagc tcaaagcaca caaggataaa tccgcacaat gttttaagat
     2161 gttttacaaa ggggttatta cccatgatgt gtctagcgct atcaatcgcc ctcagattgg
     2221 cgtggtgaga gagtttttga cgcgaaatcc cgcctggcga aaggccgtat tcatcagtcc
     2281 ttacaactcg caaaatgccg tcgctagtaa gattttagga ctccctactc agacagttga
     2341 tagctctcaa ggttctgaat acgattatgt gatttttact cagactaccg aaacggcaca
     2401 ttcttgtaac gtcaaccgtt ttaacgtggc cattacaagg gccaaggttg gcatcttgtg
     2461 tatcatgtca gaccgggacc tgtatgataa actgcaattc acatctttgg agataccgcg
     2521 ccgcaatgtc gctacccttc aggaaaacct ttactttcag tcgatggatt ataaggacca
     2581 cgatggagac tacaaagatc atgacattga ttacaaggac gacgatgaca aataagctag
     2641 cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat acattgatga
     2701 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     2761 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     2821 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     2881 cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc ccttggggcc
     2941 tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat ctaccggcat
     3001 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
     3061 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
     3121 aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg atcaagacat
     3181 tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc aaactttctt
     3241 ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca gtcaagtcag
     3301 ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg tcagtcaagt
     3361 tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt aaatcatgtg
     3421 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     3481 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     3541 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3601 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3661 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3721 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3781 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3841 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3901 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3961 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4021 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4081 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4141 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     4201 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     4261 tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc cgaactctcc
     4321 aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag gctacctctc
     4381 gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct tggcagcgcc
     4441 tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg agagaagttc
     4501 aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat cggggcgcgc
     4561 ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc
     4621 agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc
     4681 aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt
     4741 caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag
     4801 gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4861 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4921 tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4981 ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     5041 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     5101 atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     5161 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     5221 caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     5281 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     5341 ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta
     5401 ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac
     5461 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     5521 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     5581 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     5641 taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa
     5701 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     5761 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga
     5821 tcccgaatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc
     5881 gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt
     5941 acctttttgg cagcgatcgc ggctcccgac atcttggacc attagctcca caggtatctt
     6001 cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa
     6061 gacccgttta gaggccccaa gggg
//
