LOCUS       sequence-295291-        6051 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 6051)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6051)
  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-m4134462
FEATURES             Location/Qualifiers
     source          1..6051
                     /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..1867
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    1920..2041
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1931..1950)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1985..2004
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2156..2202
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2232..2368
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2413..2484
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2710..3298)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2790..2809)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4044..4112,4113..4904)
                     /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(4262..4281)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4917..5053
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5205..5276
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5281..5327)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5345..5393
                     /note="synthetic polyadenylation signal"
     enhancer        5474..5777
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5779..5981
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5932..5952
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6050..6051,1..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 agtcggggaa acctataccc aatccacttc tcggcttaga
     1861 ttctacctaa gctagcttga ctgactgaga tacagcgtac cttcagctca cagacatgat
     1921 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     1981 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     2041 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     2101 ttaaagcaag taaaacctct acaaatgtgg tattggccca tctctatcgg tatcgtagca
     2161 taaccccttg gggcctctaa acgggtcttg aggggttttt tgtgcccctc gggccggatt
     2221 gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
     2281 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
     2341 gtcctcctta ccagaaattt atccttaagg tcgtcagcta tcctgcaggc gatctctcga
     2401 tttcgatcaa gacattcctt taatggtctt ttctggacac cactaggggt cagaagtagt
     2461 tcatcaaact ttcttccctc cctaatctca ttggttacct tgggctatcg aaacttaatt
     2521 aaccagtcaa gtcagctact tggcgagatc gacttgtctg ggtttcgact acgctcagaa
     2581 ttgcgtcagt caagttcgat ctggtccttg ctattgcacc cgttctccga ttacgagttt
     2641 catttaaatc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2701 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2761 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2821 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2881 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2941 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3001 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3061 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3121 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3181 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3241 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3301 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3361 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3421 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3481 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcatttaa
     3541 atttccgaac tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc cgatccatct
     3601 tgcaggctac ctctcgaacg aactatcgca agtctcttgg ccggccttgc gccttggcta
     3661 ttgcttggca gcgcctatcg ccaggtatta ctccaatccc gaatatccga gatcgggatc
     3721 acccgagaga agttcaacct acatcctcaa tcccgatcta tccgagatcc gaggaatatc
     3781 gaaatcgggg cgcgcctggt gtaccgagaa cgatcctctc agtgcgagtc tcgacgatcc
     3841 atatcgttgc ttggcagtca gccagtcgga atccagcttg ggacccagga agtccaatcg
     3901 tcagatattg tactcaagcc tggtcacggc agcgtaccga tctgtttaaa cctagatatt
     3961 gatagtctga tcggtcaacg tataatcgag tcctagcttt tgcaaacatc tatcaagaga
     4021 caggatcagc aggaggcttt cgcatgagta ttcaacattt ccgtgtcgcc cttattccct
     4081 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4141 atgctgaaga tcagttgggt gcgcgagtgg gttacatcga actggatctc aacagcggta
     4201 agatccttga gagttttcgc cccgaagaac gctttccaat gatgagcact tttaaagttc
     4261 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     4321 tacactattc tcagaatgac ttggttgagt attcaccagt cacagaaaag catcttacgg
     4381 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     4441 ccaacttact tctgacaacg attggaggac cgaaggagct aaccgctttt ttgcacaaca
     4501 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     4561 acgacgagcg tgacaccacg atgcctgtag caatggcaac aaccttgcgt aaactattaa
     4621 ctggcgaact acttactcta gcttcccggc aacagttgat agactggatg gaggcggata
     4681 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4741 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     4801 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     4861 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaaccgatt ctaggtgcat
     4921 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
     4981 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
     5041 aatttatcct taagatcccg aatcgtttaa actcgactct ggctctatcg aatctccgtc
     5101 gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc gaatctcgtc
     5161 agctatcgtc agcttacctt tttggcagcg atcgcggctc ccgacatctt ggaccattag
     5221 ctccacaggt atcttcttcc ctctagtggt cataacagca gcttcagcta cctctcaatt
     5281 caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctatca atcgttgcgt
     5341 tacacacaca aaaaaccaac acacatccat cttcgatgga tagcgatttt attatctaac
     5401 tgctgatcga gtgtagccag atctagtaat caattacggg gtcattagtt catagcccat
     5461 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     5521 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     5581 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     5641 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     5701 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     5761 tcatcgctat taccatgctg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     5821 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     5881 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     5941 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
     6001 tcagatcttt gtcgatccta ccatccactc gacacacccg ccagcggccg c
//
