LOCUS       sequence-295727-        6102 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 6102)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6102)
  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-m4134468
FEATURES             Location/Qualifiers
     source          1..6102
                     /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             1877..1894
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1971..2092
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1982..2001)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2036..2055
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2207..2253
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2283..2419
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2464..2535
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2761..3349)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2841..2860)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4095..4163,4164..4955)
                     /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(4313..4332)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4968..5104
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5256..5327
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5332..5378)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5396..5444
                     /note="synthetic polyadenylation signal"
     enhancer        5525..5828
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5830..6032
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5983..6003
                     /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 ttcagtcgat gagtgacaaa atcattcatc taaccgatga
     1561 ctcctttgat acagacgtgt tgaaggccga tggtgcgata ctggtagatt tctgggctga
     1621 atggtgtggc ccgtgcaaga tgatcgcacc cattctggac gagatcgccg acgagtatca
     1681 aggaaaatta actgtcgcta agcttaatat agatcagaac ccagggacgg cacctaaata
     1741 cggcatcagg ggaataccta ccctcctgct ctttaagaat ggggaagttg ccgcgacaaa
     1801 agtgggagcc ctgtctaagg gccagctaaa agagttcttg gacgctaacc tggcaggttc
     1861 aggctcgggg cacatgcacc atcaccatca ccatagcagc gggctcgtgc cacgcggcta
     1921 agctagcttg actgactgag atacagcgta ccttcagctc acagacatga taagatacat
     1981 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     2041 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     2101 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaagcaa
     2161 gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc ataacccctt
     2221 ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat tgctatctac
     2281 cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
     2341 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
     2401 accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg atttcgatca
     2461 agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag ttcatcaaac
     2521 tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat taaccagtca
     2581 agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga attgcgtcag
     2641 tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt tcatttaaat
     2701 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2761 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2821 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2881 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2941 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3001 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3061 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3121 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3181 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3241 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3301 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3361 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3421 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3481 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3541 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta aatttccgaa
     3601 ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc ttgcaggcta
     3661 cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct attgcttggc
     3721 agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat cacccgagag
     3781 aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat cgaaatcggg
     3841 gcgcgcctgg tgtaccgaga acgatcctct cagtgcgagt ctcgacgatc catatcgttg
     3901 cttggcagtc agccagtcgg aatccagctt gggacccagg aagtccaatc gtcagatatt
     3961 gtactcaagc ctggtcacgg cagcgtaccg atctgtttaa acctagatat tgatagtctg
     4021 atcggtcaac gtataatcga gtcctagctt ttgcaaacat ctatcaagag acaggatcag
     4081 caggaggctt tcgcatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4141 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     4201 atcagttggg tgcgcgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     4261 agagttttcg ccccgaagaa cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg
     4321 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     4381 ctcagaatga cttggttgag tattcaccag tcacagaaaa gcatcttacg gatggcatga
     4441 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     4501 ttctgacaac gattggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     4561 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     4621 gtgacaccac gatgcctgta gcaatggcaa caaccttgcg taaactatta actggcgaac
     4681 tacttactct agcttcccgg caacagttga tagactggat ggaggcggat aaagttgcag
     4741 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     4801 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     4861 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     4921 ctgagatagg tgcctcactg attaagcatt ggtaaccgat tctaggtgca ttggcgcaga
     4981 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
     5041 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
     5101 ttaagatccc gaatcgttta aactcgactc tggctctatc gaatctccgt cgtttcgagc
     5161 ttacgcgaac agccgtggcg ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt
     5221 cagcttacct ttttggcagc gatcgcggct cccgacatct tggaccatta gctccacagg
     5281 tatcttcttc cctctagtgg tcataacagc agcttcagct acctctcaat tcaaaaaacc
     5341 cctcaagacc cgtttagagg ccccaagggg ttatgctatc aatcgttgcg ttacacacac
     5401 aaaaaaccaa cacacatcca tcttcgatgg atagcgattt tattatctaa ctgctgatcg
     5461 agtgtagcca gatctagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     5521 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     5581 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     5641 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     5701 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     5761 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     5821 ttaccatgct gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     5881 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     5941 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     6001 gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag atcagatctt
     6061 tgtcgatcct accatccact cgacacaccc gccagcggcc gc
//
