LOCUS       sequence-300166-        6831 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 6831)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6831)
  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-m4252959
FEATURES             Location/Qualifiers
     source          1..6831
                     /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..295
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    364..485
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(375..394)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     429..448
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      600..646
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      676..812
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    857..928
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1154..1742)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1234..1253)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2242..3451
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3288..3307
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3455..4483
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4510..4558
                     /note="synthetic polyadenylation signal"
     CDS             join(4824..4892,4893..5684)
                     /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(5042..5061)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5697..5833
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5985..6056
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6061..6107)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6125..6173
                     /note="synthetic polyadenylation signal"
     enhancer        6254..6557
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6559..6761
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6712..6732
                     /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 aaagaaaccg cggcagctaa gttcgagagg cagcacatgg acagttaagg
      301 atccgcaggc ctctgctagc ttgactgact gagatacagc gtaccttcag ctcacagaca
      361 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
      421 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
      481 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
      541 ttttttaaag caagtaaaac ctctacaaat gtggtattgg cccatctcta tcggtatcgt
      601 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgtgcc cctcgggccg
      661 gattgctatc taccggcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta
      721 tactcccaca tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat
      781 acgtgtcctc cttaccagaa atttatcctt aaggtcgtca gctatcctgc aggcgatctc
      841 tcgatttcga tcaagacatt cctttaatgg tcttttctgg acaccactag gggtcagaag
      901 tagttcatca aactttcttc cctccctaat ctcattggtt accttgggct atcgaaactt
      961 aattaaccag tcaagtcagc tacttggcga gatcgacttg tctgggtttc gactacgctc
     1021 agaattgcgt cagtcaagtt cgatctggtc cttgctattg cacccgttct ccgattacga
     1081 gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1141 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1201 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     1261 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     1321 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     1381 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     1441 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     1501 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     1561 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     1621 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     1681 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     1741 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     1801 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     1861 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     1921 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcat
     1981 ttaaatttcc gaactctcca aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc
     2041 atcttgcagg ctacctctcg aacgaactat cgcaagtctc ttggccggcc ttgcgccttg
     2101 gctattgctt ggcagcgcct atcgccaggt attactccaa tcccgaatat ccgagatcgg
     2161 gatcacccga gagaagttca acctacatcc tcaatcccga tctatccgag atccgaggaa
     2221 tatcgaaatc ggggcgcgcc tggcctccgc gccgggtttt ggcgcctccc gcgggcgccc
     2281 ccctcgtcac ggcgagcgct gccacgtcag acgaagggcg caggagcgtc ctgatccttc
     2341 cgcccggacg ctcaggacag cggcccgctg ctcataagac tcggccttag aaccccagta
     2401 tcagcagaag gacattttag gacgggactt gggtgactct agggcactgg ttttctttcc
     2461 agagagcgga acaggcgagg aaaagtagtc ccttctcggc gattctgcgg agggatctcc
     2521 gtggggcggt gaacgccgat gattatataa ggacgcgccg ggtgtggcac agctagttcc
     2581 gtcgcagccg ggatttgggt cgcggttctt gtttgtggat cgctgtgatc gtcacttggt
     2641 gagtagcggg ctgctgggct ggccggggct ttcgtggccg ccgggccgct cggtgggacg
     2701 gaagcgtgtg gagagaccgc caagggctgt agtctgggtc cgcgagcaag gttgccctga
     2761 actgggggtt ggggggagcg cagcaaaatg gcggctgttc ccgagtcttg aatggaagac
     2821 gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga
     2881 acccaaggtc ttgagccctt cgctaatgcg ggaaagctct tattcgggtg agatgggctg
     2941 ggcaccatct ggggaccctg acgtgaagtt tgtcactgac tggagaactc ggtttgtcgt
     3001 ctgttgcggg ggcggcagtt atggcggtgc cgttgggcag tgcacccgta cctttgggag
     3061 cgcgcgccct cgtcgtgtcg tgacgtcacc cgttctgttg gcttataatg cagggtgggg
     3121 ccaccagccg gtaggtgtgc ggtaggcttt tctccgtcgc aggacgcagg gttcgggcct
     3181 agggtaggct ctcctgaatc gacaggcgcc ggacctctgg tgaggggagg gataagtgag
     3241 gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt aagtagctga agctccggtt
     3301 ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa gttttttagg caccttttga
     3361 aatgtaatca tttgggtcaa tatgtaattt tcagtgttag acttgtaaat tgtccgctaa
     3421 attctggccg tttttggctt ttttgttaga caacatgggt aaaaagcctg aactcaccgc
     3481 gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct
     3541 ctcggagggc gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct
     3601 gcgggtaaat agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc
     3661 atccgccgcg ctcccgattc cggaagtgct tgacattggg gagttcagcg agagcctgac
     3721 ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact
     3781 gcccgctgtt ctgcagccgg tcgcggaggc aatggatgcc atcgctgccg ccgatcttag
     3841 ccagacgagc gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg
     3901 tgatttcata tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga
     3961 caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg
     4021 ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa
     4081 tggccgcata acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga
     4141 ggtcgccaac atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta
     4201 cttcgagcgg aggcatccgg agcttgcagg atcgccccgg ctccgggcgt atatgctccg
     4261 cattggtctt gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg
     4321 ggcgcagggt cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca
     4381 aatcgcccgc agaagcgccg ccgtctggac cgatggctgt gtagaagtac tcgccgatag
     4441 tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa taagctagta tgtaagccta
     4501 gtcttagata ataaaatcgc tatccatcga agatggatgt gtgttggttt tttgtgtgtg
     4561 taacgctagg cgcgcctggt gtaccgagaa cgatcctctc agtgcgagtc tcgacgatcc
     4621 atatcgttgc ttggcagtca gccagtcgga atccagcttg ggacccagga agtccaatcg
     4681 tcagatattg tactcaagcc tggtcacggc agcgtaccga tctgtttaaa cctagatatt
     4741 gatagtctga tcggtcaacg tataatcgag tcctagcttt tgcaaacatc tatcaagaga
     4801 caggatcagc aggaggcttt cgcatgagta ttcaacattt ccgtgtcgcc cttattccct
     4861 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4921 atgctgaaga tcagttgggt gcgcgagtgg gttacatcga actggatctc aacagcggta
     4981 agatccttga gagttttcgc cccgaagaac gctttccaat gatgagcact tttaaagttc
     5041 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     5101 tacactattc tcagaatgac ttggttgagt attcaccagt cacagaaaag catcttacgg
     5161 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5221 ccaacttact tctgacaacg attggaggac cgaaggagct aaccgctttt ttgcacaaca
     5281 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5341 acgacgagcg tgacaccacg atgcctgtag caatggcaac aaccttgcgt aaactattaa
     5401 ctggcgaact acttactcta gcttcccggc aacagttgat agactggatg gaggcggata
     5461 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5521 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5581 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5641 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaaccgatt ctaggtgcat
     5701 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
     5761 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
     5821 aatttatcct taagatcccg aatcgtttaa actcgactct ggctctatcg aatctccgtc
     5881 gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc gaatctcgtc
     5941 agctatcgtc agcttacctt tttggcagcg atcgcggctc ccgacatctt ggaccattag
     6001 ctccacaggt atcttcttcc ctctagtggt cataacagca gcttcagcta cctctcaatt
     6061 caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctatca atcgttgcgt
     6121 tacacacaca aaaaaccaac acacatccat cttcgatgga tagcgatttt attatctaac
     6181 tgctgatcga gtgtagccag atctagtaat caattacggg gtcattagtt catagcccat
     6241 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     6301 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     6361 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     6421 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     6481 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     6541 tcatcgctat taccatgctg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     6601 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     6661 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     6721 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga
     6781 tcagatcttt gtcgatccta ccatccactc gacacacccg ccagcggccg c
//
