LOCUS       sequence-297410-        6410 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 6410)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6410)
  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-m4252785
FEATURES             Location/Qualifiers
     source          1..6410
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             230..250
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             254..319
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    372..493
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(383..402)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     437..456
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      608..654
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      684..820
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    865..936
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1162..1750)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1242..1261)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2250..3459
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3296..3315
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3463..4062
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3463..3482)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3959..3979
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4089..4137
                     /note="synthetic polyadenylation signal"
     CDS             join(4403..4471,4472..5263)
                     /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(4621..4640)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5276..5412
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5564..5635
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5640..5686)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5704..5752
                     /note="synthetic polyadenylation signal"
     enhancer        5833..6136
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6138..6340
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6291..6311
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgtac tcattcgttt cggaagagac aggtacgtta atagttaata gcgtacttct
       61 ttttcttgct ttcgtggtat tcttgctagt tacactagcc atccttactg cgcttcgatt
      121 gtgtgcgtac tgctgcaata ttgttaacgt gagtcttgta aaaccttctt tttacgttta
      181 ctctcgtgtt aaaaatctga attcttctag agttcctgat cttctggtcg aaaaccttta
      241 ctttcagtcg atggattata aggaccacga tggagactac aaagatcatg acattgatta
      301 caaggacgac gatgacaaat aagctagctt gactgactga gatacagcgt accttcagct
      361 cacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
      421 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
      481 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
      541 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtattggcc catctctatc
      601 ggtatcgtag cataacccct tggggcctct aaacgggtct tgaggggttt tttgtgcccc
      661 tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc
      721 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc
      781 acttccatac gtgtcctcct taccagaaat ttatccttaa ggtcgtcagc tatcctgcag
      841 gcgatctctc gatttcgatc aagacattcc tttaatggtc ttttctggac accactaggg
      901 gtcagaagta gttcatcaaa ctttcttccc tccctaatct cattggttac cttgggctat
      961 cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga tcgacttgtc tgggtttcga
     1021 ctacgctcag aattgcgtca gtcaagttcg atctggtcct tgctattgca cccgttctcc
     1081 gattacgagt ttcatttaaa tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     1141 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     1201 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     1261 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     1321 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     1381 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     1441 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     1501 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     1561 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     1621 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     1681 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     1741 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     1801 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     1861 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     1921 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     1981 agttgcattt aaatttccga actctccaag gccctcgtcg gaaaatcttc aaacctttcg
     2041 tccgatccat cttgcaggct acctctcgaa cgaactatcg caagtctctt ggccggcctt
     2101 gcgccttggc tattgcttgg cagcgcctat cgccaggtat tactccaatc ccgaatatcc
     2161 gagatcggga tcacccgaga gaagttcaac ctacatcctc aatcccgatc tatccgagat
     2221 ccgaggaata tcgaaatcgg ggcgcgcctg gcctccgcgc cgggttttgg cgcctcccgc
     2281 gggcgccccc ctcgtcacgg cgagcgctgc cacgtcagac gaagggcgca ggagcgtcct
     2341 gatccttccg cccggacgct caggacagcg gcccgctgct cataagactc ggccttagaa
     2401 ccccagtatc agcagaagga cattttagga cgggacttgg gtgactctag ggcactggtt
     2461 ttctttccag agagcggaac aggcgaggaa aagtagtccc ttctcggcga ttctgcggag
     2521 ggatctccgt ggggcggtga acgccgatga ttatataagg acgcgccggg tgtggcacag
     2581 ctagttccgt cgcagccggg atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt
     2641 cacttggtga gtagcgggct gctgggctgg ccggggcttt cgtggccgcc gggccgctcg
     2701 gtgggacgga agcgtgtgga gagaccgcca agggctgtag tctgggtccg cgagcaaggt
     2761 tgccctgaac tgggggttgg ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa
     2821 tggaagacgc ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg
     2881 cggcaagaac ccaaggtctt gagcccttcg ctaatgcggg aaagctctta ttcgggtgag
     2941 atgggctggg caccatctgg ggaccctgac gtgaagtttg tcactgactg gagaactcgg
     3001 tttgtcgtct gttgcggggg cggcagttat ggcggtgccg ttgggcagtg cacccgtacc
     3061 tttgggagcg cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca
     3121 gggtggggcc accagccggt aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt
     3181 tcgggcctag ggtaggctct cctgaatcga caggcgccgg acctctggtg aggggaggga
     3241 taagtgaggc gtcagtttct ttggtcggtt ttatgtacct atcttcttaa gtagctgaag
     3301 ctccggtttt gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca
     3361 ccttttgaaa tgtaatcatt tgggtcaata tgtaattttc agtgttagac ttgtaaattg
     3421 tccgctaaat tctggccgtt tttggctttt ttgttagaca acatgaccga gtacaagccc
     3481 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     3541 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     3601 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     3661 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     3721 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     3781 caacagatgg agggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     3841 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     3901 ggagtggagg ccgccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     3961 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     4021 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct aggctagtat gtaagcctag
     4081 tcttagataa taaaatcgct atccatcgaa gatggatgtg tgttggtttt ttgtgtgtgt
     4141 aacgctaggc gcgcctggtg taccgagaac gatcctctca gtgcgagtct cgacgatcca
     4201 tatcgttgct tggcagtcag ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt
     4261 cagatattgt actcaagcct ggtcacggca gcgtaccgat ctgtttaaac ctagatattg
     4321 atagtctgat cggtcaacgt ataatcgagt cctagctttt gcaaacatct atcaagagac
     4381 aggatcagca ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4441 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4501 tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa ctggatctca acagcggtaa
     4561 gatccttgag agttttcgcc ccgaagaacg ctttccaatg atgagcactt ttaaagttct
     4621 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4681 acactattct cagaatgact tggttgagta ttcaccagtc acagaaaagc atcttacgga
     4741 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4801 caacttactt ctgacaacga ttggaggacc gaaggagcta accgcttttt tgcacaacat
     4861 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4921 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca accttgcgta aactattaac
     4981 tggcgaacta cttactctag cttcccggca acagttgata gactggatgg aggcggataa
     5041 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5101 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5161 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5221 acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc taggtgcatt
     5281 ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat
     5341 tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa
     5401 atttatcctt aagatcccga atcgtttaaa ctcgactctg gctctatcga atctccgtcg
     5461 tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgggcatcg aatctcgtca
     5521 gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg gaccattagc
     5581 tccacaggta tcttcttccc tctagtggtc ataacagcag cttcagctac ctctcaattc
     5641 aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt
     5701 acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta ttatctaact
     5761 gctgatcgag tgtagccaga tctagtaatc aattacgggg tcattagttc atagcccata
     5821 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     5881 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     5941 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     6001 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     6061 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     6121 catcgctatt accatgctga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     6181 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     6241 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     6301 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
     6361 cagatctttg tcgatcctac catccactcg acacacccgc cagcggccgc
//
