LOCUS       sequence-307369-        6855 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 6855)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6855)
  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-m4252962
FEATURES             Location/Qualifiers
     source          1..6855
                     /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    388..509
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(399..418)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     453..472
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      624..670
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      700..836
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    881..952
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1178..1766)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1258..1277)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2266..3475
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3312..3331
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3479..4507
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4534..4582
                     /note="synthetic polyadenylation signal"
     CDS             join(4848..4916,4917..5708)
                     /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(5066..5085)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5721..5857
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6009..6080
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6085..6131)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6149..6197
                     /note="synthetic polyadenylation signal"
     enhancer        6278..6581
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6583..6785
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6736..6756
                     /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 atggattata aggaccacga tggagactac aaagatcatg acattgatta
      301 caaggacgac gatgacaaat aaggatccgc aggcctctgc tagcttgact gactgagata
      361 cagcgtacct tcagctcaca gacatgataa gatacattga tgagtttgga caaaccacaa
      421 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
      481 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
      541 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
      601 ttggcccatc tctatcggta tcgtagcata accccttggg gcctctaaac gggtcttgag
      661 gggttttttg tgcccctcgg gccggattgc tatctaccgg cattggcgca gaaaaaaatg
      721 cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg
      781 cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaaggtc
      841 gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga cattccttta atggtctttt
      901 ctggacacca ctaggggtca gaagtagttc atcaaacttt cttccctccc taatctcatt
      961 ggttaccttg ggctatcgaa acttaattaa ccagtcaagt cagctacttg gcgagatcga
     1021 cttgtctggg tttcgactac gctcagaatt gcgtcagtca agttcgatct ggtccttgct
     1081 attgcacccg ttctccgatt acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa
     1141 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     1201 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     1261 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     1321 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     1381 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     1441 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     1501 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     1561 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     1621 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     1681 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     1741 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     1801 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     1861 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     1921 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     1981 tatttcgttc atccatagtt gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa
     2041 atcttcaaac ctttcgtccg atccatcttg caggctacct ctcgaacgaa ctatcgcaag
     2101 tctcttggcc ggccttgcgc cttggctatt gcttggcagc gcctatcgcc aggtattact
     2161 ccaatcccga atatccgaga tcgggatcac ccgagagaag ttcaacctac atcctcaatc
     2221 ccgatctatc cgagatccga ggaatatcga aatcggggcg cgcctggcct ccgcgccggg
     2281 ttttggcgcc tcccgcgggc gcccccctcg tcacggcgag cgctgccacg tcagacgaag
     2341 ggcgcaggag cgtcctgatc cttccgcccg gacgctcagg acagcggccc gctgctcata
     2401 agactcggcc ttagaacccc agtatcagca gaaggacatt ttaggacggg acttgggtga
     2461 ctctagggca ctggttttct ttccagagag cggaacaggc gaggaaaagt agtcccttct
     2521 cggcgattct gcggagggat ctccgtgggg cggtgaacgc cgatgattat ataaggacgc
     2581 gccgggtgtg gcacagctag ttccgtcgca gccgggattt gggtcgcggt tcttgtttgt
     2641 ggatcgctgt gatcgtcact tggtgagtag cgggctgctg ggctggccgg ggctttcgtg
     2701 gccgccgggc cgctcggtgg gacggaagcg tgtggagaga ccgccaaggg ctgtagtctg
     2761 ggtccgcgag caaggttgcc ctgaactggg ggttgggggg agcgcagcaa aatggcggct
     2821 gttcccgagt cttgaatgga agacgcttgt gaggcgggct gtgaggtcgt tgaaacaagg
     2881 tggggggcat ggtgggcggc aagaacccaa ggtcttgagc ccttcgctaa tgcgggaaag
     2941 ctcttattcg ggtgagatgg gctgggcacc atctggggac cctgacgtga agtttgtcac
     3001 tgactggaga actcggtttg tcgtctgttg cgggggcggc agttatggcg gtgccgttgg
     3061 gcagtgcacc cgtacctttg ggagcgcgcg ccctcgtcgt gtcgtgacgt cacccgttct
     3121 gttggcttat aatgcagggt ggggccacca gccggtaggt gtgcggtagg cttttctccg
     3181 tcgcaggacg cagggttcgg gcctagggta ggctctcctg aatcgacagg cgccggacct
     3241 ctggtgaggg gagggataag tgaggcgtca gtttctttgg tcggttttat gtacctatct
     3301 tcttaagtag ctgaagctcc ggttttgaac tatgcgctcg gggttggcga gtgtgttttg
     3361 tgaagttttt taggcacctt ttgaaatgta atcatttggg tcaatatgta attttcagtg
     3421 ttagacttgt aaattgtccg ctaaattctg gccgtttttg gcttttttgt tagacaacat
     3481 gggtaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga
     3541 cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga
     3601 tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga
     3661 tcgttatgtt tatcggcact ttgcatccgc cgcgctcccg attccggaag tgcttgacat
     3721 tggggagttc agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt
     3781 gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggcaatgga
     3841 tgccatcgct gccgccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg
     3901 aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta
     3961 tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga
     4021 gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg
     4081 ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc
     4141 gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc
     4201 ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc
     4261 ccggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga
     4321 cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg
     4381 agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gccgccgtct ggaccgatgg
     4441 ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa
     4501 ggaataagct agtatgtaag cctagtctta gataataaaa tcgctatcca tcgaagatgg
     4561 atgtgtgttg gttttttgtg tgtgtaacgc taggcgcgcc tggtgtaccg agaacgatcc
     4621 tctcagtgcg agtctcgacg atccatatcg ttgcttggca gtcagccagt cggaatccag
     4681 cttgggaccc aggaagtcca atcgtcagat attgtactca agcctggtca cggcagcgta
     4741 ccgatctgtt taaacctaga tattgatagt ctgatcggtc aacgtataat cgagtcctag
     4801 cttttgcaaa catctatcaa gagacaggat cagcaggagg ctttcgcatg agtattcaac
     4861 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4921 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcgcga gtgggttaca
     4981 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgctttc
     5041 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     5101 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtattcac
     5161 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     5221 taaccatgag tgataacact gcggccaact tacttctgac aacgattgga ggaccgaagg
     5281 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     5341 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     5401 caacaacctt gcgtaaacta ttaactggcg aactacttac tctagcttcc cggcaacagt
     5461 tgatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     5521 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     5581 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     5641 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5701 attggtaacc gattctaggt gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
     5761 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
     5821 tccatacgtg tcctccttac cagaaattta tccttaagat cccgaatcgt ttaaactcga
     5881 ctctggctct atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt
     5941 gctcgtcggg catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg
     6001 gctcccgaca tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac
     6061 agcagcttca gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag
     6121 gggttatgct atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga
     6181 tggatagcga ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta
     6241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     6301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     6361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     6421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     6481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     6541 cttggcagta catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt
     6601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     6661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     6721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     6781 gagctggttt agtgaaccgt cagatcagat ctttgtcgat cctaccatcc actcgacaca
     6841 cccgccagcg gccgc
//
