LOCUS       sequence-295190-        7334 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 7334)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7334)
  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-m4133293
FEATURES             Location/Qualifiers
     source          1..7334
                     /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             284..361
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             389..409
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    874..995
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(885..904)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     939..958
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1110..1156
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1186..1322
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1367..1438
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1664..2252)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1744..1763)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2752..3961
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3798..3817
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3965..4993
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5020..5068
                     /note="synthetic polyadenylation signal"
     CDS             join(5334..5402,5403..6194)
                     /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(5552..5571)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6207..6343
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6488..6559
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6564..6610)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6628..6676
                     /note="synthetic polyadenylation signal"
     enhancer        6757..7060
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7062..7264
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7215..7235
                     /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 ggacggcgaa tcccggggct gataaatccc tggaaaagga gatggaagaa
      301 gaactttatt gcagtgtctg cggctaaccg gttcaaaaag atcagttcaa gcggcgccct
      361 agattacgac attcctacga ccgcctccga gaatctctat ttccagggtg aatttggctt
      421 ggcacaacat gacgaggccg tcgataacaa gtttaataaa gaacagcaga acgctttcta
      481 tgagatcctt cacttaccaa atctgaacga agagcaacgc aacgctttca ttcagagcct
      541 gaaagacgat ccctcgcagt ctgcaaatct cctggccgaa gccaagaagc tgaatgacgc
      601 acaggctcca aaagttgata acaagtttaa caaggagcag cagaatgctt tctacgagat
      661 attgcacctt cctaatctga atgaggaaca aagaaatgca tttatccagt ccctaaaaga
      721 cgatccatca cagagtgcca acctgcttgc ggaagccaaa aagctcaatg atgcccaggc
      781 cccgaaagta gacgcgaacc atcaataagg atccgcaggc ctctgctagc ttgactgact
      841 gagatacagc gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa
      901 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
      961 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     1021 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
     1081 gtggtattgg cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt
     1141 cttgaggggt tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa
     1201 aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg
     1261 atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt
     1321 aaggtcgtca gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg
     1381 tcttttctgg acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat
     1441 ctcattggtt accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga
     1501 gatcgacttg tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc
     1561 cttgctattg cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc
     1621 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     1681 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     1741 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     1801 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     1861 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     1921 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     1981 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     2041 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     2101 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     2161 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     2221 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     2281 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     2341 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     2401 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     2461 tctgtctatt tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt
     2521 cggaaaatct tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat
     2581 cgcaagtctc ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt
     2641 attactccaa tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc
     2701 tcaatcccga tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggcctccgc
     2761 gccgggtttt ggcgcctccc gcgggcgccc ccctcgtcac ggcgagcgct gccacgtcag
     2821 acgaagggcg caggagcgtc ctgatccttc cgcccggacg ctcaggacag cggcccgctg
     2881 ctcataagac tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt
     2941 gggtgactct agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc
     3001 ccttctcggc gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatataa
     3061 ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt
     3121 gtttgtggat cgctgtgatc gtcacttggt gagtagcggg ctgctgggct ggccggggct
     3181 ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc caagggctgt
     3241 agtctgggtc cgcgagcaag gttgccctga actgggggtt ggggggagcg cagcaaaatg
     3301 gcggctgttc ccgagtcttg aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa
     3361 acaaggtggg gggcatggtg ggcggcaaga acccaaggtc ttgagccctt cgctaatgcg
     3421 ggaaagctct tattcgggtg agatgggctg ggcaccatct ggggaccctg acgtgaagtt
     3481 tgtcactgac tggagaactc ggtttgtcgt ctgttgcggg ggcggcagtt atggcggtgc
     3541 cgttgggcag tgcacccgta cctttgggag cgcgcgccct cgtcgtgtcg tgacgtcacc
     3601 cgttctgttg gcttataatg cagggtgggg ccacctgccg gtaggtgtgc ggtaggcttt
     3661 tctccgtcgc aggacgcagg gttcgggcct agggtaggct ctcctgaatc gacaggcgcc
     3721 ggacctctgg tgaggggagg gataagtgag gcgtcagttt ctttggtcgg ttttatgtac
     3781 ctatcttctt aagtagctga agctccggtt ttgaactatg cgctcggggt tggcgagtgt
     3841 gttttgtgaa gttttttagg caccttttga aatgtaatca tttgggtcaa tatgtaattt
     3901 tcagtgttag acttgtaaat tgtccgctaa attctggccg tttttggctt ttttgttaga
     3961 caacatgggt aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa
     4021 gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag
     4081 cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta
     4141 caaagatcgt tatgtttatc ggcactttgc atccgccgcg ctcccgattc cggaagtgct
     4201 tgacattggg gagttcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt
     4261 cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc
     4321 aatggatgcc atcgctgccg ccgatcttag ccagacgagc gggttcggcc cattcggacc
     4381 gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca
     4441 tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct
     4501 cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga
     4561 tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag
     4621 cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg
     4681 gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg
     4741 atcgccccgg ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt
     4801 ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg
     4861 atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgccg ccgtctggac
     4921 cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag
     4981 ggcaaaggaa taagctagta tgtaagccta gtcttagata ataaaatcgc tatccatcga
     5041 agatggatgt gtgttggttt tttgtgtgtg taacgctagg cgcgcctggt gtaccgagaa
     5101 cgatcctctc agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga
     5161 atccagcttg ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc
     5221 agcgtaccga tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag
     5281 tcctagcttt tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta
     5341 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     5401 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg
     5461 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5521 gctttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5581 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5641 attcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5701 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac
     5761 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5821 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5881 caatggcaac aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc
     5941 aacagttgat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     6001 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     6061 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     6121 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     6181 ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc
     6241 tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg
     6301 cccacttcca tacgtgtcct ccttaccaga aatttatcct taagatcgtt taaactcgac
     6361 tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg cgctcatttg
     6421 ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca gcgatcgcgg
     6481 ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt ggtcataaca
     6541 gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga ggccccaagg
     6601 ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc catcttcgat
     6661 ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt aatcaattac
     6721 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     6781 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     6841 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     6901 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     6961 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     7021 ttggcagtac atctacgtat tagtcatcgc tattaccatg ctgatgcggt tttggcagta
     7081 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     7141 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     7201 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     7261 agctggttta gtgaaccgtc agatcagatc tttgtcgatc ctaccatcca ctcgacacac
     7321 ccgccagcgg ccgc
//
