LOCUS       sequence-297382-        6383 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 6383)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6383)
  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-m4252828
FEATURES             Location/Qualifiers
     source          1..6383
                     /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             251..292
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    345..466
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(356..375)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     410..429
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      581..627
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      657..793
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    838..909
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1135..1723)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1215..1234)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2223..3432
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3269..3288
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3436..4035
                     /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(3436..3455)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3932..3952
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4062..4110
                     /note="synthetic polyadenylation signal"
     CDS             join(4376..4444,4445..5236)
                     /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(4594..4613)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5249..5385
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5537..5608
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5613..5659)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5677..5725
                     /note="synthetic polyadenylation signal"
     enhancer        5806..6109
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6111..6313
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6264..6284
                     /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 gggaaaccta tacccaatcc acttctcggc ttagattcta cctaagctag
      301 cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat acattgatga
      361 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
      421 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
      481 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
      541 cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc ccttggggcc
      601 tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat ctaccggcat
      661 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
      721 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
      781 aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg atcaagacat
      841 tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc aaactttctt
      901 ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca gtcaagtcag
      961 ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg tcagtcaagt
     1021 tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt aaatcatgtg
     1081 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     1141 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     1201 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1261 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1321 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     1381 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     1441 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     1501 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     1561 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     1621 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     1681 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     1741 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     1801 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     1861 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     1921 tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc cgaactctcc
     1981 aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag gctacctctc
     2041 gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct tggcagcgcc
     2101 tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg agagaagttc
     2161 aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat cggggcgcgc
     2221 ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc cccctcgtca cggcgagcgc
     2281 tgccacgtca gacgaagggc gcaggagcgt cctgatcctt ccgcccggac gctcaggaca
     2341 gcggcccgct gctcataaga ctcggcctta gaaccccagt atcagcagaa ggacatttta
     2401 ggacgggact tgggtgactc tagggcactg gttttctttc cagagagcgg aacaggcgag
     2461 gaaaagtagt cccttctcgg cgattctgcg gagggatctc cgtggggcgg tgaacgccga
     2521 tgattatata aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc gggatttggg
     2581 tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc
     2641 tggccggggc tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg
     2701 ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg aactgggggt tggggggagc
     2761 gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg
     2821 aggtcgttga aacaaggtgg ggggcatggt gggcggcaag aacccaaggt cttgagccct
     2881 tcgctaatgc gggaaagctc ttattcgggt gagatgggct gggcaccatc tggggaccct
     2941 gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg gggcggcagt
     3001 tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc
     3061 gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccaccagcc ggtaggtgtg
     3121 cggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc tctcctgaat
     3181 cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt tctttggtcg
     3241 gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat gcgctcgggg
     3301 ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca
     3361 atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc gtttttggct
     3421 tttttgttag acaacatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     3481 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     3541 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     3601 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     3661 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     3721 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggagggcct cctggcgccg
     3781 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     3841 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggccgccga gcgcgccggg
     3901 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     3961 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     4021 aagcccggtg cctaggctag tatgtaagcc tagtcttaga taataaaatc gctatccatc
     4081 gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta ggcgcgcctg gtgtaccgag
     4141 aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt cagccagtcg
     4201 gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag cctggtcacg
     4261 gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa cgtataatcg
     4321 agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct ttcgcatgag
     4381 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4441 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt
     4501 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4561 acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4621 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4681 gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4741 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgattggagg
     4801 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4861 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     4921 agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc tagcttcccg
     4981 gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5041 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5101 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5161 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5221 gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac
     5281 gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact
     5341 tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcc cgaatcgttt
     5401 aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc
     5461 gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc tttttggcag
     5521 cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt ccctctagtg
     5581 gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag
     5641 gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca acacacatcc
     5701 atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc agatctagta
     5761 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     5821 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     5881 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     5941 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     6001 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     6061 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc tgatgcggtt
     6121 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     6181 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     6241 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     6301 tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc taccatccac
     6361 tcgacacacc cgccagcggc cgc
//
