LOCUS       sequence-295857-        6564 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 6564)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6564)
  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-m4134394
FEATURES             Location/Qualifiers
     source          1..6564
                     /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             1859..1936
                     /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"
     CDS             1964..1984
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    2433..2554
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2444..2463)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2498..2517
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2669..2715
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2745..2881
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2926..2997
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3223..3811)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3303..3322)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4557..4625,4626..5417)
                     /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(4775..4794)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5430..5566
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5718..5789
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5794..5840)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5858..5906
                     /note="synthetic polyadenylation signal"
     enhancer        5987..6290
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6292..6494
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6445..6465
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6563..6564,1..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 agtcgggacg gcgaatcccg gggctgataa atccctggaa
     1861 aaggagatgg aagaagaact ttattgcagt gtctgcggct aaccggttca aaaagatcag
     1921 ttcaagcggc gccctagatt acgacattcc tacgaccgcc tccgagaatc tctatttcca
     1981 gggtgaattt ggcttggcac aacatgacga ggccgtcgat aacaagttta ataaagaaca
     2041 gcagaacgct ttctatgaga tccttcactt accaaatctg aacgaagagc aacgcaacgc
     2101 tttcattcag agcctgaaag acgatccctc gcagtctgca aatctcctgg ccgaagccaa
     2161 gaagctgaat gacgcacagg ctccaaaagt tgataacaag tttaacaagg agcagcagaa
     2221 tgctttctac gagatattgc accttcctaa tctgaatgag gaacaaagaa atgcatttat
     2281 ccagtcccta aaagacgatc catcacagag tgccaacctg cttgcggaag ccaaaaagct
     2341 caatgatgcc caggccccga aagtagacgc gaaccatcaa taagctagct tgactgactg
     2401 agatacagcg taccttcagc tcacagacat gataagatac attgatgagt ttggacaaac
     2461 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     2521 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     2581 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     2641 tggtattggc ccatctctat cggtatcgta gcataacccc ttggggcctc taaacgggtc
     2701 ttgaggggtt ttttgtgccc ctcgggccgg attgctatct accggcattg gcgcagaaaa
     2761 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     2821 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     2881 aggtcgtcag ctatcctgca ggcgatctct cgatttcgat caagacattc ctttaatggt
     2941 cttttctgga caccactagg ggtcagaagt agttcatcaa actttcttcc ctccctaatc
     3001 tcattggtta ccttgggcta tcgaaactta attaaccagt caagtcagct acttggcgag
     3061 atcgacttgt ctgggtttcg actacgctca gaattgcgtc agtcaagttc gatctggtcc
     3121 ttgctattgc acccgttctc cgattacgag tttcatttaa atcatgtgag caaaaggcca
     3181 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3241 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3301 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3361 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3421 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3481 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3541 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3601 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3661 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3721 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3781 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3841 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3901 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3961 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4021 ctgtctattt cgttcatcca tagttgcatt taaatttccg aactctccaa ggccctcgtc
     4081 ggaaaatctt caaacctttc gtccgatcca tcttgcaggc tacctctcga acgaactatc
     4141 gcaagtctct tggccggcct tgcgccttgg ctattgcttg gcagcgccta tcgccaggta
     4201 ttactccaat cccgaatatc cgagatcggg atcacccgag agaagttcaa cctacatcct
     4261 caatcccgat ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct ggtgtaccga
     4321 gaacgatcct ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc
     4381 ggaatccagc ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac
     4441 ggcagcgtac cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc
     4501 gagtcctagc ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga
     4561 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     4621 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag
     4681 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     4741 aacgctttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     4801 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     4861 agtattcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     4921 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgattggag
     4981 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     5041 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     5101 tagcaatggc aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc
     5161 ggcaacagtt gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     5221 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     5281 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     5341 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     5401 tgattaagca ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga
     5461 cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac
     5521 ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaagatc ccgaatcgtt
     5581 taaactcgac tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg
     5641 cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca
     5701 gcgatcgcgg ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt
     5761 ggtcataaca gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga
     5821 ggccccaagg ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc
     5881 catcttcgat ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt
     5941 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     6001 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     6061 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     6121 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     6181 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     6241 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg ctgatgcggt
     6301 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     6361 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     6421 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     6481 atataagcag agctggttta gtgaaccgtc agatcagatc tttgtcgatc ctaccatcca
     6541 ctcgacacac ccgccagcgg ccgc
//
