LOCUS       sequence-300728-        4476 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 E (envelope).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4476)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4476)
  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-m4133220
FEATURES             Location/Qualifiers
     source          1..4476
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    138..186
                     /note="synthetic polyadenylation signal"
     terminator      204..250
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    255..326
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(478..614)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(627..1418,1419..1487))
                     /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     1250..1269
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2233..2821
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2722..2741
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    3047..3118
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3163..3299)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(3329..3375)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3490..3611
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3527..3546)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3581..3600
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(3664..3705)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(3706..3726)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(4026..4228)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4055..4075)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        join(4230..4476,1..57)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg
       61 aactccatat atgggctatg aactaatgac cccgtaattg attactagat ctggctacac
      121 tcgatcagca gttagataat aaaatcgcta tccatcgaag atggatgtgt gttggttttt
      181 tgtgtgtgta acgcaacgat tgatagcata accccttggg gcctctaaac gggtcttgag
      241 gggttttttg aattgagagg tagctgaagc tgctgttatg accactagag ggaagaagat
      301 acctgtggag ctaatggtcc aagatgtcgg gagccgcgat cgctgccaaa aaggtaagct
      361 gacgatagct gacgagattc gatgcccgac gagcaaatga gcgccacggc tgttcgcgta
      421 agctcgaaac gacggagatt cgatagagcc agagtcgagt ttaaacgatt cgggatctta
      481 aggataaatt tctggtaagg aggacacgta tggaagtggg caagttgggg aagccgtatc
      541 cgttgctgaa tctggcatat gtgggagtat aagacgcgca gcgtcgcatc aggcattttt
      601 ttctgcgcca atgcacctag aatcggttac caatgcttaa tcagtgaggc acctatctca
      661 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
      721 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
      781 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
      841 cctgcaactt tatccgcctc catccagtct atcaactgtt gccgggaagc tagagtaagt
      901 agttcgccag ttaatagttt acgcaaggtt gttgccattg ctacaggcat cgtggtgtca
      961 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     1021 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccaat cgttgtcaga
     1081 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     1141 gtcatgccat ccgtaagatg cttttctgtg actggtgaat actcaaccaa gtcattctga
     1201 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     1261 ccacatagca gaactttaaa agtgctcatc attggaaagc gttcttcggg gcgaaaactc
     1321 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgcgc acccaactga
     1381 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     1441 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatgcg aaagcctcct
     1501 gctgatcctg tctcttgata gatgtttgca aaagctagga ctcgattata cgttgaccga
     1561 tcagactatc aatatctagg tttaaacaga tcggtacgct gccgtgacca ggcttgagta
     1621 caatatctga cgattggact tcctgggtcc caagctggat tccgactggc tgactgccaa
     1681 gcaacgatat ggatcgtcga gactcgcact gagaggatcg ttctcggtac accaggcgcg
     1741 ccccgatttc gatattcctc ggatctcgga tagatcggga ttgaggatgt aggttgaact
     1801 tctctcgggt gatcccgatc tcggatattc gggattggag taatacctgg cgataggcgc
     1861 tgccaagcaa tagccaaggc gcaaggccgg ccaagagact tgcgatagtt cgttcgagag
     1921 gtagcctgca agatggatcg gacgaaaggt ttgaagattt tccgacgagg gccttggaga
     1981 gttcggaaat ttaaatgcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     2041 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     2101 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     2161 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     2221 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2281 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2341 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     2401 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2461 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     2521 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     2581 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     2641 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     2701 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     2761 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     2821 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     2881 gatttaaatg aaactcgtaa tcggagaacg ggtgcaatag caaggaccag atcgaacttg
     2941 actgacgcaa ttctgagcgt agtcgaaacc cagacaagtc gatctcgcca agtagctgac
     3001 ttgactggtt aattaagttt cgatagccca aggtaaccaa tgagattagg gagggaagaa
     3061 agtttgatga actacttctg acccctagtg gtgtccagaa aagaccatta aaggaatgtc
     3121 ttgatcgaaa tcgagagatc gcctgcagga tagctgacga ccttaaggat aaatttctgg
     3181 taaggaggac acgtatggaa gtgggcaagt tggggaagcc gtatccgttg ctgaatctgg
     3241 catatgtggg agtataagac gcgcagcgtc gcatcaggca tttttttctg cgccaatgcc
     3301 ggtagatagc aatccggccc gaggggcaca aaaaacccct caagacccgt ttagaggccc
     3361 caaggggtta tgctacgata ccgatagaga tgggccaata ccacatttgt agaggtttta
     3421 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     3481 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3541 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3601 aatgtatctt atcatgtctg tgagctgaag gtacgctgta tctcagtcag tcaagctagc
     3661 ttaggtagaa tctaagccga gaagtggatt gggtataggt ttccccgact gaaagtaaag
     3721 gttttcgacc agaagatcag gaactctaga agaattcaga tttttaacac gagagtaaac
     3781 gtaaaaagaa ggttttacaa gactcacgtt aacaatattg cagcagtacg cacacaatcg
     3841 aagcgcagta aggatggcta gtgtaactag caagaatacc acgaaagcaa gaaaaagaag
     3901 tacgctatta actattaacg tacctgtctc ttccgaaacg aatgagtaca tggtggcggc
     3961 cgctggcggg tgtgtcgagt ggatggtagg atcgacaaag atctgatctg acggttcact
     4021 aaaccagctc tgcttatata gacctcccac cgtacacgcc taccgcccat ttgcgtcaat
     4081 ggggcggagt tgttacgaca ttttggaaag tcccgttgat tttggtgcca aaacaaactc
     4141 ccattgacgt caatggggtg gagacttgga aatccccgtg agtcaaaccg ctatccacgc
     4201 ccattgatgt actgccaaaa ccgcatcagc atggtaatag cgatgactaa tacgtagatg
     4261 tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat
     4321 ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc
     4381 aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc
     4441 gttactatgg gaacatacgt cattattgac gtcaat
//
