LOCUS       sequence-300224-        6036 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 6036)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6036)
  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-m4134333
FEATURES             Location/Qualifiers
     source          1..6036
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      257..393
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    545..616
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(621..667)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    685..733
                     /note="synthetic polyadenylation signal"
     enhancer        814..1117
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1119..1321
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1272..1292
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1390..1399
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3196..3216
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3217..3243
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3296..3417
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3307..3326)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3361..3380
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      3532..3578
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3608..3744
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3789..3860
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4086..4674)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4166..4185)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(5420..5488,5489..6036,1..244)
                     /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(5638..5657)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
       61 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
      121 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
      181 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
      241 gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt
      301 atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca
      361 tacgtgtcct ccttaccaga aatttatcct taagatcccg aatcgtttaa actcgactct
      421 ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc
      481 gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg atcgcggctc
      541 ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt cataacagca
      601 gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt
      661 tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat cttcgatgga
      721 tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat caattacggg
      781 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      841 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      901 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      961 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     1021 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     1081 gcagtacatc tacgtattag tcatcgctat taccatgctg atgcggtttt ggcagtacat
     1141 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     1201 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     1261 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     1321 tggtttagtg aaccgtcaga tcagatcttt gtcgatccta ccatccactc gacacacccg
     1381 ccagcggccg ccaccatggt tggggcttgt gttctttgca attcacagac ttcattaaga
     1441 tgtggtgctt gcatacgtag accattctta tgttgtaaat gctgttacga ccatgtcata
     1501 tcaacatcac ataaattagt cttgtctgtt aatccgtatg tttgcaatgc tccaggttgt
     1561 gatgtcacag atgtgactca actttactta ggaggtatga gctattattg taaatcacat
     1621 aaaccaccca ttagttttcc attgtgtgct aatggacaag tttttggttt atataaaaat
     1681 acatgtgttg gtagcgataa tgttactgac tttaatgcaa ttgcaacatg tgactggaca
     1741 aatgctggtg attacatttt agctaacacc tgtactgaaa gactcaagct ttttgcagca
     1801 gaaacgctca aagctactga ggagacattt aaactgtctt atggtattgc tactgtacgt
     1861 gaagtgctgt ctgacagaga attacatctt tcatgggaag ttggtaaacc tagaccacca
     1921 cttaaccgaa attatgtctt tactggttat cgtgtaacta aaaacagtaa agtacaaata
     1981 ggagagtaca cctttgaaaa aggtgactat ggtgatgctg ttgtttaccg aggtacaaca
     2041 acttacaaat taaatgttgg tgattatttt gtgctgacat cacatacagt aatgccatta
     2101 agtgcaccta cactagtgcc acaagagcac tatgttagaa ttactggctt atacccaaca
     2161 ctcaatatct cagatgagtt ttctagcaat gttgcaaatt atcaaaaggt tggtatgcaa
     2221 aagtattcta cactccaggg accacctggt actggtaaga gtcattttgc tattggccta
     2281 gctctctact acccttctgc tcgcatagtg tatacagctt gctctcatgc cgctgttgat
     2341 gcactatgtg agaaggcatt aaaatatttg cctatagata aatgtagtag aattatacct
     2401 gcacgtgctc gtgtagagtg ttttgataaa ttcaaagtga attcaacatt agaacagtat
     2461 gtcttttgta ctgtaaatgc attgcctgag acgacagcag atatagttgt ctttgatgaa
     2521 atttcaatgg ccacaaatta tgatttgagt gttgtcaatg ccagattacg tgctaagcac
     2581 tatgtgtaca ttggcgaccc tgctcaatta cctgcaccac gcacattgct aactaagggc
     2641 acactagaac cagaatattt caattcagtg tgtagactta tgaaaactat aggtccagac
     2701 atgttcctcg gaacttgtcg gcgttgtcct gctgaaattg ttgacactgt gagtgctttg
     2761 gtttatgata ataagcttaa agcacataaa gacaaatcag ctcaatgctt taaaatgttt
     2821 tataagggtg ttatcacgca tgatgtttca tctgcaatta acaggccaca aataggcgtg
     2881 gtaagagaat tccttacacg taaccctgct tggagaaaag ctgtctttat ttcaccttat
     2941 aattcacaga atgctgtagc ctcaaagatt ttgggactac caactcaaac tgttgattca
     3001 tcacagggct cagaatatga ctatgtcata ttcactcaaa ccactgaaac agctcactct
     3061 tgtaatgtaa acagatttaa tgttgctatt accagagcaa aagtaggcat actttgcata
     3121 atgtctgata gagaccttta tgacaagttg caatttacaa gtcttgaaat tccacgtagg
     3181 aatgtggcaa ctttagaaaa cctttacttt cagtcgtacc cctatgacgt accagattat
     3241 gcgtaagcta gcttgactga ctgagataca gcgtaccttc agctcacaga catgataaga
     3301 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     3361 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     3421 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     3481 agcaagtaaa acctctacaa atgtggtatt ggcccatctc tatcggtatc gtagcataac
     3541 cccttggggc ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc cggattgcta
     3601 tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca
     3661 catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc
     3721 tccttaccag aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc
     3781 gatcaagaca ttcctttaat ggtcttttct ggacaccact aggggtcaga agtagttcat
     3841 caaactttct tccctcccta atctcattgg ttaccttggg ctatcgaaac ttaattaacc
     3901 agtcaagtca gctacttggc gagatcgact tgtctgggtt tcgactacgc tcagaattgc
     3961 gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt ctccgattac gagtttcatt
     4021 taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4081 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4141 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4201 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4261 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4321 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4381 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4441 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4501 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4561 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4621 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4681 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4741 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4801 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     4861 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc atttaaattt
     4921 ccgaactctc caaggccctc gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca
     4981 ggctacctct cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct tggctattgc
     5041 ttggcagcgc ctatcgccag gtattactcc aatcccgaat atccgagatc gggatcaccc
     5101 gagagaagtt caacctacat cctcaatccc gatctatccg agatccgagg aatatcgaaa
     5161 tcggggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga cgatccatat
     5221 cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc caatcgtcag
     5281 atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta gatattgata
     5341 gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc aagagacagg
     5401 atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta ttcccttttt
     5461 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     5521 tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca gcggtaagat
     5581 ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta aagttctgct
     5641 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     5701 ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc ttacggatgg
     5761 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     5821 cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5881 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     5941 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg
     6001 cgaactactt actctagctt cccggcaaca gttgat
//
