LOCUS       sequence-295559-        5757 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 nsp4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5757)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5757)
  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-m4133820
FEATURES             Location/Qualifiers
     source          1..5757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1508..1528
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1529..1573
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    1626..1747
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1637..1656)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1691..1710
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1862..1908
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1938..2074
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2119..2190
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2416..3004)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2496..2515)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3750..3818,3819..4610)
                     /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(3968..3987)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4623..4759
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4911..4982
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4987..5033)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5051..5099
                     /note="synthetic polyadenylation signal"
     enhancer        5180..5483
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5485..5687
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5638..5658
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgaag attgtaaata actggttgaa acaactaata aaggttactt tagtgttcct
       61 gtttgtcgcc gcaatcttct atcttatcac cccggtgcac gtgatgagta aacatacaga
      121 cttttcttca gaaattatcg ggtacaaggc tatcgatggc ggtgtaacgc gcgacattgc
      181 ctccacagat acctgcttcg cgaacaaaca cgctgacttt gatacttggt tcagccagcg
      241 aggaggctcg tacaccaatg acaaagcctg tccactgatc gcagccgtga taactagaga
      301 ggttggattc gtcgtccctg ggctccccgg gacaattctc cgtacaacca acggcgactt
      361 tctgcatttt ctgccccggg tgttctccgc agtcggtaat atctgttata ctccatctaa
      421 gctcattgag tacacggatt ttgctaccag cgcgtgcgtg ttggccgcag aatgcaccat
      481 atttaaggac gcttctggaa aacctgtgcc ctattgttac gataccaatg tgcttgaggg
      541 ctcagttgcc tatgaaagtc tgaggccaga cactagatac gttttaatgg atggaagcat
      601 catccagttc ccaaacacgt atctggaggg gtccgtgcgg gtcgtgacca cattcgattc
      661 agaatactgc cgccacggta catgcgagag gtctgaggct ggcgtatgtg tgtcaactag
      721 cggtcgatgg gttctaaata acgattatta tcgttctctg cccggcgttt tttgtggagt
      781 ggacgccgtt aaccttttga ccaacatgtt tacacctctc attcagccca ttggtgcact
      841 ggatatctcc gcgagtattg tggctggggg aatagtcgcc attgtcgtaa cctgcttggc
      901 atactatttc atgcgtttca ggcgagcttt tggcgaatac tcacatgtcg tggccttcaa
      961 tactcttctg tttctgatga gcttcacggt actctgtctg accccagtat acagtttcct
     1021 acctggggtg tattctgtga tctacttata cctcacattt tacctgacca atgacgtgtc
     1081 ctttctggcc cacatccaat ggatggttat gttcacgccg ctcgtgcctt tctggattac
     1141 aatcgcttat ataatctgta ttagcacgaa gcatttctat tggttctttt ccaactactt
     1201 aaagcggcgc gtcgtattta atggggtatc gtttagcact tttgaggaag ccgcgctctg
     1261 cactttcttg cttaataagg agatgtattt gaaattaaga tctgacgttc tgcttccttt
     1321 aacacagtac aacagatacc tggcgttgta taacaaatac aaatatttct cgggagccat
     1381 ggatacaact tcctatagag aggctgcttg ttgccacctt gcaaaggcac tcaacgactt
     1441 cagtaacagc ggcagtgatg tgctttatca gccccctcag acttcaataa catccgccgt
     1501 cctccaagaa aacctttact ttcagtcgaa agaaaccgcg gcagctaagt tcgagaggca
     1561 gcacatggac agttaagcta gcttgactga ctgagataca gcgtaccttc agctcacaga
     1621 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     1681 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     1741 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     1801 ggttttttaa agcaagtaaa acctctacaa atgtggtatt ggcccatctc tatcggtatc
     1861 gtagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc
     1921 cggattgcta tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
     1981 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
     2041 atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc
     2101 tctcgatttc gatcaagaca ttcctttaat ggtcttttct ggacaccact aggggtcaga
     2161 agtagttcat caaactttct tccctcccta atctcattgg ttaccttggg ctatcgaaac
     2221 ttaattaacc agtcaagtca gctacttggc gagatcgact tgtctgggtt tcgactacgc
     2281 tcagaattgc gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt ctccgattac
     2341 gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2401 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2461 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2521 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2581 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2641 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2701 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2761 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2821 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2881 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2941 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3001 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3061 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3121 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3181 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3241 atttaaattt ccgaactctc caaggccctc gtcggaaaat cttcaaacct ttcgtccgat
     3301 ccatcttgca ggctacctct cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct
     3361 tggctattgc ttggcagcgc ctatcgccag gtattactcc aatcccgaat atccgagatc
     3421 gggatcaccc gagagaagtt caacctacat cctcaatccc gatctatccg agatccgagg
     3481 aatatcgaaa tcggggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga
     3541 cgatccatat cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc
     3601 caatcgtcag atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta
     3661 gatattgata gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc
     3721 aagagacagg atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta
     3781 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     3841 taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca
     3901 gcggtaagat ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta
     3961 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     4021 gccgcataca ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc
     4081 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     4141 ctgcggccaa cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc
     4201 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     4261 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac
     4321 tattaactgg cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg
     4381 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     4441 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     4501 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     4561 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag
     4621 gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
     4681 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
     4741 accagaaatt tatccttaag atcccgaatc gtttaaactc gactctggct ctatcgaatc
     4801 tccgtcgttt cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat
     4861 ctcgtcagct atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac
     4921 cattagctcc acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc
     4981 tcaattcaaa aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg
     5041 ttgcgttaca cacacaaaaa accaacacac atccatcttc gatggatagc gattttatta
     5101 tctaactgct gatcgagtgt agccagatct agtaatcaat tacggggtca ttagttcata
     5161 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     5221 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     5281 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     5341 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     5401 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     5461 tattagtcat cgctattacc atgctgatgc ggttttggca gtacatcaat gggcgtggat
     5521 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     5581 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     5641 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
     5701 gtcagatcag atctttgtcg atcctaccat ccactcgaca cacccgccag cggccgc
//
