LOCUS       sequence-307376-        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"
     primer_bind     259..278
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             426..1025
                     /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(426..445)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     922..942
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    1052..1100
                     /note="synthetic polyadenylation signal"
     CDS             join(1366..1434,1435..2226)
                     /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(1584..1603)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      2239..2375
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2527..2598
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2603..2649)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    2667..2715
                     /note="synthetic polyadenylation signal"
     enhancer        2796..3099
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3101..3303
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3254..3274
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3603..3623
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3627..3692
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    3745..3866
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3756..3775)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3810..3829
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      3981..4027
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4057..4193
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4238..4309
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(4535..5123)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4615..4634)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(5623..6410,1..422)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
ORIGIN
        1 ccgttgggca gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac
       61 ccgttctgtt ggcttataat gcagggtggg gccaccagcc ggtaggtgtg cggtaggctt
      121 ttctccgtcg caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc
      181 cggacctctg gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta
      241 cctatcttct taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg
      301 tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt
      361 ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag
      421 acaacatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg
      481 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc
      541 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
      601 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
      661 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
      721 gcggttcccg gctggccgcg cagcaacaga tggagggcct cctggcgccg caccggccca
      781 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
      841 tgggcagcgc cgtcgtgctc cccggagtgg aggccgccga gcgcgccggg gtgcccgcct
      901 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
      961 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
     1021 cctaggctag tatgtaagcc tagtcttaga taataaaatc gctatccatc gaagatggat
     1081 gtgtgttggt tttttgtgtg tgtaacgcta ggcgcgcctg gtgtaccgag aacgatcctc
     1141 tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt cagccagtcg gaatccagct
     1201 tgggacccag gaagtccaat cgtcagatat tgtactcaag cctggtcacg gcagcgtacc
     1261 gatctgttta aacctagata ttgatagtct gatcggtcaa cgtataatcg agtcctagct
     1321 tttgcaaaca tctatcaaga gacaggatca gcaggaggct ttcgcatgag tattcaacat
     1381 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     1441 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt gggttacatc
     1501 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgctttcca
     1561 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     1621 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtattcacca
     1681 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     1741 accatgagtg ataacactgc ggccaactta cttctgacaa cgattggagg accgaaggag
     1801 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     1861 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     1921 acaaccttgc gtaaactatt aactggcgaa ctacttactc tagcttcccg gcaacagttg
     1981 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     2041 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     2101 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     2161 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     2221 tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc
     2281 ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc
     2341 catacgtgtc ctccttacca gaaatttatc cttaagatcc cgaatcgttt aaactcgact
     2401 ctggctctat cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc
     2461 tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc tttttggcag cgatcgcggc
     2521 tcccgacatc ttggaccatt agctccacag gtatcttctt ccctctagtg gtcataacag
     2581 cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag gccccaaggg
     2641 gttatgctat caatcgttgc gttacacaca caaaaaacca acacacatcc atcttcgatg
     2701 gatagcgatt ttattatcta actgctgatc gagtgtagcc agatctagta atcaattacg
     2761 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     2821 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     2881 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     2941 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     3001 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     3061 tggcagtaca tctacgtatt agtcatcgct attaccatgc tgatgcggtt ttggcagtac
     3121 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
     3181 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
     3241 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
     3301 gctggtttag tgaaccgtca gatcagatct ttgtcgatcc taccatccac tcgacacacc
     3361 cgccagcggc cgccaccatg tactcattcg tttcggaaga gacaggtacg ttaatagtta
     3421 atagcgtact tctttttctt gctttcgtgg tattcttgct agttacacta gccatcctta
     3481 ctgcgcttcg attgtgtgcg tactgctgca atattgttaa cgtgagtctt gtaaaacctt
     3541 ctttttacgt ttactctcgt gttaaaaatc tgaattcttc tagagttcct gatcttctgg
     3601 tcgaaaacct ttactttcag tcgatggatt ataaggacca cgatggagac tacaaagatc
     3661 atgacattga ttacaaggac gacgatgaca aataagctag cttgactgac tgagatacag
     3721 cgtaccttca gctcacagac atgataagat acattgatga gtttggacaa accacaacta
     3781 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     3841 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     3901 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg
     3961 gcccatctct atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg
     4021 ttttttgtgc ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct
     4081 gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt
     4141 ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc
     4201 agctatcctg caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg
     4261 gacaccacta ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt
     4321 taccttgggc tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt
     4381 gtctgggttt cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt
     4441 gcacccgttc tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg
     4501 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4561 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4621 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4681 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4741 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4801 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4861 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4921 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     4981 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5041 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5101 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5161 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5221 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5281 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5341 ttcgttcatc catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc
     5401 ttcaaacctt tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct
     5461 cttggccggc cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca
     5521 atcccgaata tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg
     5581 atctatccga gatccgagga atatcgaaat cggggcgcgc ctggcctccg cgccgggttt
     5641 tggcgcctcc cgcgggcgcc cccctcgtca cggcgagcgc tgccacgtca gacgaagggc
     5701 gcaggagcgt cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga
     5761 ctcggcctta gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc
     5821 tagggcactg gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg
     5881 cgattctgcg gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc
     5941 gggtgtggca cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga
     6001 tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc
     6061 gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt
     6121 ccgcgagcaa ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt
     6181 cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg
     6241 ggggcatggt gggcggcaag aacccaaggt cttgagccct tcgctaatgc gggaaagctc
     6301 ttattcgggt gagatgggct gggcaccatc tggggaccct gacgtgaagt ttgtcactga
     6361 ctggagaact cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg
//
