LOCUS       sequence-297477-        6804 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 6804)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6804)
  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-m4252945
FEATURES             Location/Qualifiers
     source          1..6804
                     /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..268
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    337..458
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(348..367)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     402..421
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      573..619
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      649..785
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    830..901
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1127..1715)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1207..1226)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2215..3424
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3261..3280
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3428..4456
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4483..4531
                     /note="synthetic polyadenylation signal"
     CDS             join(4797..4865,4866..5657)
                     /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(5015..5034)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5670..5806
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5958..6029
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6034..6080)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6098..6146
                     /note="synthetic polyadenylation signal"
     enhancer        6227..6530
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6532..6734
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6685..6705
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgtac agctttgtat ctgaagagac aggcactctc atcgttaatt cagtgttgct
       61 gttcctggca tttgtggtct tccttttagt gaccctagcg atactgacgg ccctccggct
      121 gtgcgcttat tgttgtaaca ttgtcaacgt tagtctcgtg aagccctcgt tctacgttta
      181 ttcccgcgtg aaaaatctaa acagctcccg tgtgcctgat cttcttgtcg aaaaccttta
      241 ctttcagtcg caccatcacc atcaccatta aggatccgca ggcctctgct agcttgactg
      301 actgagatac agcgtacctt cagctcacag acatgataag atacattgat gagtttggac
      361 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
      421 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
      481 ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca
      541 aatgtggtat tggcccatct ctatcggtat cgtagcataa ccccttgggg cctctaaacg
      601 ggtcttgagg ggttttttgt gcccctcggg ccggattgct atctaccggc attggcgcag
      661 aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa
      721 cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc
      781 cttaaggtcg tcagctatcc tgcaggcgat ctctcgattt cgatcaagac attcctttaa
      841 tggtcttttc tggacaccac taggggtcag aagtagttca tcaaactttc ttccctccct
      901 aatctcattg gttaccttgg gctatcgaaa cttaattaac cagtcaagtc agctacttgg
      961 cgagatcgac ttgtctgggt ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg
     1021 gtccttgcta ttgcacccgt tctccgatta cgagtttcat ttaaatcatg tgagcaaaag
     1081 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     1141 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     1201 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     1261 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     1321 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     1381 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     1441 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     1501 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     1561 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     1621 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     1681 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     1741 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     1801 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     1861 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     1921 cgatctgtct atttcgttca tccatagttg catttaaatt tccgaactct ccaaggccct
     1981 cgtcggaaaa tcttcaaacc tttcgtccga tccatcttgc aggctacctc tcgaacgaac
     2041 tatcgcaagt ctcttggccg gccttgcgcc ttggctattg cttggcagcg cctatcgcca
     2101 ggtattactc caatcccgaa tatccgagat cgggatcacc cgagagaagt tcaacctaca
     2161 tcctcaatcc cgatctatcc gagatccgag gaatatcgaa atcggggcgc gcctggcctc
     2221 cgcgccgggt tttggcgcct cccgcgggcg cccccctcgt cacggcgagc gctgccacgt
     2281 cagacgaagg gcgcaggagc gtcctgatcc ttccgcccgg acgctcagga cagcggcccg
     2341 ctgctcataa gactcggcct tagaacccca gtatcagcag aaggacattt taggacggga
     2401 cttgggtgac tctagggcac tggttttctt tccagagagc ggaacaggcg aggaaaagta
     2461 gtcccttctc ggcgattctg cggagggatc tccgtggggc ggtgaacgcc gatgattata
     2521 taaggacgcg ccgggtgtgg cacagctagt tccgtcgcag ccgggatttg ggtcgcggtt
     2581 cttgtttgtg gatcgctgtg atcgtcactt ggtgagtagc gggctgctgg gctggccggg
     2641 gctttcgtgg ccgccgggcc gctcggtggg acggaagcgt gtggagagac cgccaagggc
     2701 tgtagtctgg gtccgcgagc aaggttgccc tgaactgggg gttgggggga gcgcagcaaa
     2761 atggcggctg ttcccgagtc ttgaatggaa gacgcttgtg aggcgggctg tgaggtcgtt
     2821 gaaacaaggt ggggggcatg gtgggcggca agaacccaag gtcttgagcc cttcgctaat
     2881 gcgggaaagc tcttattcgg gtgagatggg ctgggcacca tctggggacc ctgacgtgaa
     2941 gtttgtcact gactggagaa ctcggtttgt cgtctgttgc gggggcggca gttatggcgg
     3001 tgccgttggg cagtgcaccc gtacctttgg gagcgcgcgc cctcgtcgtg tcgtgacgtc
     3061 acccgttctg ttggcttata atgcagggtg gggccaccag ccggtaggtg tgcggtaggc
     3121 ttttctccgt cgcaggacgc agggttcggg cctagggtag gctctcctga atcgacaggc
     3181 gccggacctc tggtgagggg agggataagt gaggcgtcag tttctttggt cggttttatg
     3241 tacctatctt cttaagtagc tgaagctccg gttttgaact atgcgctcgg ggttggcgag
     3301 tgtgttttgt gaagtttttt aggcaccttt tgaaatgtaa tcatttgggt caatatgtaa
     3361 ttttcagtgt tagacttgta aattgtccgc taaattctgg ccgtttttgg cttttttgtt
     3421 agacaacatg ggtaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga
     3481 aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt
     3541 cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt
     3601 ctacaaagat cgttatgttt atcggcactt tgcatccgcc gcgctcccga ttccggaagt
     3661 gcttgacatt ggggagttca gcgagagcct gacctattgc atctcccgcc gtgcacaggg
     3721 tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga
     3781 ggcaatggat gccatcgctg ccgccgatct tagccagacg agcgggttcg gcccattcgg
     3841 accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc
     3901 ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc
     3961 tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc
     4021 ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg
     4081 gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc
     4141 gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc
     4201 aggatcgccc cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag
     4261 cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt
     4321 ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg ccgccgtctg
     4381 gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc
     4441 gagggcaaag gaataagcta gtatgtaagc ctagtcttag ataataaaat cgctatccat
     4501 cgaagatgga tgtgtgttgg ttttttgtgt gtgtaacgct aggcgcgcct ggtgtaccga
     4561 gaacgatcct ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc
     4621 ggaatccagc ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac
     4681 ggcagcgtac cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc
     4741 gagtcctagc ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga
     4801 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     4861 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag
     4921 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     4981 aacgctttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     5041 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     5101 agtattcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     5161 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgattggag
     5221 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     5281 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     5341 tagcaatggc aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc
     5401 ggcaacagtt gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     5461 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     5521 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     5581 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     5641 tgattaagca ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga
     5701 cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac
     5761 ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaagatc ccgaatcgtt
     5821 taaactcgac tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg
     5881 cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca
     5941 gcgatcgcgg ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt
     6001 ggtcataaca gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga
     6061 ggccccaagg ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc
     6121 catcttcgat ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt
     6181 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     6241 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     6301 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     6361 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     6421 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     6481 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg ctgatgcggt
     6541 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     6601 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     6661 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     6721 atataagcag agctggttta gtgaaccgtc agatcagatc tttgtcgatc ctaccatcca
     6781 ctcgacacac ccgccagcgg ccgc
//
