LOCUS       sequence-295498-        5754 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 5754)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5754)
  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-m4133973
FEATURES             Location/Qualifiers
     source          1..5754
                     /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..1570
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    1623..1744
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1634..1653)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1688..1707
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1859..1905
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1935..2071
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2116..2187
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2413..3001)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2493..2512)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3747..3815,3816..4607)
                     /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(3965..3984)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4620..4756
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4908..4979
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4984..5030)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5048..5096
                     /note="synthetic polyadenylation signal"
     enhancer        5177..5480
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5482..5684
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5635..5655
                     /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 ttcagtcggg gaaacctata cccaatccac ttctcggctt
     1561 agattctacc taagctagct tgactgactg agatacagcg taccttcagc tcacagacat
     1621 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     1681 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     1741 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     1801 tttttaaagc aagtaaaacc tctacaaatg tggtattggc ccatctctat cggtatcgta
     1861 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc ctcgggccgg
     1921 attgctatct accggcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat
     1981 actcccacat atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata
     2041 cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag ctatcctgca ggcgatctct
     2101 cgatttcgat caagacattc ctttaatggt cttttctgga caccactagg ggtcagaagt
     2161 agttcatcaa actttcttcc ctccctaatc tcattggtta ccttgggcta tcgaaactta
     2221 attaaccagt caagtcagct acttggcgag atcgacttgt ctgggtttcg actacgctca
     2281 gaattgcgtc agtcaagttc gatctggtcc ttgctattgc acccgttctc cgattacgag
     2341 tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2401 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2461 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2521 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2581 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2641 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2701 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2761 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2821 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     2881 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2941 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3001 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3061 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3121 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3181 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcatt
     3241 taaatttccg aactctccaa ggccctcgtc ggaaaatctt caaacctttc gtccgatcca
     3301 tcttgcaggc tacctctcga acgaactatc gcaagtctct tggccggcct tgcgccttgg
     3361 ctattgcttg gcagcgccta tcgccaggta ttactccaat cccgaatatc cgagatcggg
     3421 atcacccgag agaagttcaa cctacatcct caatcccgat ctatccgaga tccgaggaat
     3481 atcgaaatcg gggcgcgcct ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga
     3541 tccatatcgt tgcttggcag tcagccagtc ggaatccagc ttgggaccca ggaagtccaa
     3601 tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac cgatctgttt aaacctagat
     3661 attgatagtc tgatcggtca acgtataatc gagtcctagc ttttgcaaac atctatcaag
     3721 agacaggatc agcaggaggc tttcgcatga gtattcaaca tttccgtgtc gcccttattc
     3781 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     3841 aagatgctga agatcagttg ggtgcgcgag tgggttacat cgaactggat ctcaacagcg
     3901 gtaagatcct tgagagtttt cgccccgaag aacgctttcc aatgatgagc acttttaaag
     3961 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     4021 gcatacacta ttctcagaat gacttggttg agtattcacc agtcacagaa aagcatctta
     4081 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     4141 cggccaactt acttctgaca acgattggag gaccgaagga gctaaccgct tttttgcaca
     4201 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     4261 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaaccttg cgtaaactat
     4321 taactggcga actacttact ctagcttccc ggcaacagtt gatagactgg atggaggcgg
     4381 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     4441 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     4501 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     4561 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaaccg attctaggtg
     4621 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
     4681 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
     4741 agaaatttat ccttaagatc ccgaatcgtt taaactcgac tctggctcta tcgaatctcc
     4801 gtcgtttcga gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc
     4861 gtcagctatc gtcagcttac ctttttggca gcgatcgcgg ctcccgacat cttggaccat
     4921 tagctccaca ggtatcttct tccctctagt ggtcataaca gcagcttcag ctacctctca
     4981 attcaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta tcaatcgttg
     5041 cgttacacac acaaaaaacc aacacacatc catcttcgat ggatagcgat tttattatct
     5101 aactgctgat cgagtgtagc cagatctagt aatcaattac ggggtcatta gttcatagcc
     5161 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     5221 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     5281 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     5341 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     5401 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     5461 tagtcatcgc tattaccatg ctgatgcggt tttggcagta catcaatggg cgtggatagc
     5521 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     5581 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     5641 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
     5701 agatcagatc tttgtcgatc ctaccatcca ctcgacacac ccgccagcgg ccgc
//
