LOCUS       sequence-295747-        6087 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 6087)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6087)
  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-m4133135
FEATURES             Location/Qualifiers
     source          1..6087
                     /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..1903
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     polyA_signal    1956..2077
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1967..1986)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2021..2040
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2192..2238
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2268..2404
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2449..2520
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2746..3334)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2826..2845)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4080..4148,4149..4940)
                     /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(4298..4317)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4953..5089
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5241..5312
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5317..5363)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5381..5429
                     /note="synthetic polyadenylation signal"
     enhancer        5510..5813
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5815..6017
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5968..5988
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6086..6087,1..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 agtcgaagag gcgatggaaa aagaatttta tcgcggtgag
     1861 tgcagccaac agattcaaaa agatatcctc gtctggtgcc ctctaagcta gcttgactga
     1921 ctgagataca gcgtaccttc agctcacaga catgataaga tacattgatg agtttggaca
     1981 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     2041 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     2101 tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa
     2161 atgtggtatt ggcccatctc tatcggtatc gtagcataac cccttggggc ctctaaacgg
     2221 gtcttgaggg gttttttgtg cccctcgggc cggattgcta tctaccggca ttggcgcaga
     2281 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
     2341 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
     2401 ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc gatcaagaca ttcctttaat
     2461 ggtcttttct ggacaccact aggggtcaga agtagttcat caaactttct tccctcccta
     2521 atctcattgg ttaccttggg ctatcgaaac ttaattaacc agtcaagtca gctacttggc
     2581 gagatcgact tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg
     2641 tccttgctat tgcacccgtt ctccgattac gagtttcatt taaatcatgt gagcaaaagg
     2701 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2761 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2821 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2881 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2941 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3001 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3061 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3121 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3181 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3241 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3301 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3361 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3421 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3481 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3541 gatctgtcta tttcgttcat ccatagttgc atttaaattt ccgaactctc caaggccctc
     3601 gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca ggctacctct cgaacgaact
     3661 atcgcaagtc tcttggccgg ccttgcgcct tggctattgc ttggcagcgc ctatcgccag
     3721 gtattactcc aatcccgaat atccgagatc gggatcaccc gagagaagtt caacctacat
     3781 cctcaatccc gatctatccg agatccgagg aatatcgaaa tcggggcgcg cctggtgtac
     3841 cgagaacgat cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca
     3901 gtcggaatcc agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt
     3961 cacggcagcg taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata
     4021 atcgagtcct agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgca
     4081 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4141 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc
     4201 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4261 aagaacgctt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     4321 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     4381 ttgagtattc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     4441 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgattg
     4501 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     4561 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     4621 ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg cgaactactt actctagctt
     4681 cccggcaaca gttgatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     4741 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     4801 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     4861 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     4921 cactgattaa gcattggtaa ccgattctag gtgcattggc gcagaaaaaa atgcctgatg
     4981 cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc
     5041 aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag atcccgaatc
     5101 gtttaaactc gactctggct ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg
     5161 tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct tacctttttg
     5221 gcagcgatcg cggctcccga catcttggac cattagctcc acaggtatct tcttccctct
     5281 agtggtcata acagcagctt cagctacctc tcaattcaaa aaacccctca agacccgttt
     5341 agaggcccca aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa accaacacac
     5401 atccatcttc gatggatagc gattttatta tctaactgct gatcgagtgt agccagatct
     5461 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     5521 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     5581 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     5641 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     5701 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     5761 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atgctgatgc
     5821 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     5881 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     5941 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
     6001 tctatataag cagagctggt ttagtgaacc gtcagatcag atctttgtcg atcctaccat
     6061 ccactcgaca cacccgccag cggccgc
//
