LOCUS       sequence-295683-        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"
     CDS             1805..1825
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1826..1852
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1905..2026
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1916..1935)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1970..1989
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2141..2187
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2217..2353
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2398..2469
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2695..3283)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2775..2794)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4029..4097,4098..4889)
                     /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(4247..4266)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4902..5038
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5190..5261
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5266..5312)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5330..5378
                     /note="synthetic polyadenylation signal"
     enhancer        5459..5762
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5764..5966
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5917..5937
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6035..6036,1..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 agtcgtaccc ctatgacgta ccagattatg cgtaagctag
     1861 cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat acattgatga
     1921 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     1981 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     2041 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     2101 cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc ccttggggcc
     2161 tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat ctaccggcat
     2221 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
     2281 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
     2341 aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg atcaagacat
     2401 tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc aaactttctt
     2461 ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca gtcaagtcag
     2521 ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg tcagtcaagt
     2581 tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt aaatcatgtg
     2641 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2701 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2761 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2821 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2881 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2941 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3001 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3061 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3121 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3181 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3241 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3301 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3361 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3421 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3481 tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc cgaactctcc
     3541 aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag gctacctctc
     3601 gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct tggcagcgcc
     3661 tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg agagaagttc
     3721 aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat cggggcgcgc
     3781 ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc
     3841 agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc
     3901 aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt
     3961 caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag
     4021 gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4081 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4141 tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4201 ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     4261 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     4321 atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     4381 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     4441 caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     4501 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     4561 ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta
     4621 ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac
     4681 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     4741 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     4801 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     4861 taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa
     4921 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
     4981 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga
     5041 tcccgaatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc
     5101 gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt
     5161 acctttttgg cagcgatcgc ggctcccgac atcttggacc attagctcca caggtatctt
     5221 cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa
     5281 gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa
     5341 ccaacacaca tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta
     5401 gccagatcta gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
     5461 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
     5521 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
     5581 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
     5641 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
     5701 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
     5761 tgctgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
     5821 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
     5881 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
     5941 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatcaga tctttgtcga
     6001 tcctaccatc cactcgacac acccgccagc ggccgc
//
