LOCUS       sequence-297386-        6828 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 6828)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6828)
  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-m4252873
FEATURES             Location/Qualifiers
     source          1..6828
                     /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..292
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    361..482
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(372..391)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     426..445
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      597..643
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      673..809
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    854..925
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1151..1739)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1231..1250)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2239..3448
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3285..3304
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3452..4480
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4507..4555
                     /note="synthetic polyadenylation signal"
     CDS             join(4821..4889,4890..5681)
                     /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(5039..5058)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5694..5830
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5982..6053
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6058..6104)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6122..6170
                     /note="synthetic polyadenylation signal"
     enhancer        6251..6554
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6556..6758
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6709..6729
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgtac tcattcgttt cggaagagac aggtacgtta atagttaata gcgtacttct
       61 ttttcttgct ttcgtggtat tcttgctagt tacactagcc atccttactg cgcttcgatt
      121 gtgtgcgtac tgctgcaata ttgttaacgt gagtcttgta aaaccttctt tttacgttta
      181 ctctcgtgtt aaaaatctga attcttctag agttcctgat cttctggtcg aaaaccttta
      241 ctttcagtcg gggaaaccta tacccaatcc acttctcggc ttagattcta cctaaggatc
      301 cgcaggcctc tgctagcttg actgactgag atacagcgta ccttcagctc acagacatga
      361 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
      421 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
      481 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
      541 tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc
      601 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat
      661 tgctatctac cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac
      721 tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg
      781 tgtcctcctt accagaaatt tatccttaag gtcgtcagct atcctgcagg cgatctctcg
      841 atttcgatca agacattcct ttaatggtct tttctggaca ccactagggg tcagaagtag
      901 ttcatcaaac tttcttccct ccctaatctc attggttacc ttgggctatc gaaacttaat
      961 taaccagtca agtcagctac ttggcgagat cgacttgtct gggtttcgac tacgctcaga
     1021 attgcgtcag tcaagttcga tctggtcctt gctattgcac ccgttctccg attacgagtt
     1081 tcatttaaat catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     1141 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     1201 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     1261 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     1321 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     1381 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     1441 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     1501 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     1561 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     1621 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     1681 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     1741 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     1801 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     1861 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     1921 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta
     1981 aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc
     2041 ttgcaggcta cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct
     2101 attgcttggc agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat
     2161 cacccgagag aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat
     2221 cgaaatcggg gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc
     2281 tcgtcacggc gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg atccttccgc
     2341 ccggacgctc aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca
     2401 gcagaaggac attttaggac gggacttggg tgactctagg gcactggttt tctttccaga
     2461 gagcggaaca ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg
     2521 gggcggtgaa cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc
     2581 gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag
     2641 tagcgggctg ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa
     2701 gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact
     2761 gggggttggg gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct
     2821 tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc
     2881 caaggtcttg agcccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggc
     2941 accatctggg gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg
     3001 ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc
     3061 gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct tataatgcag ggtggggcca
     3121 ccagccggta ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt cgggcctagg
     3181 gtaggctctc ctgaatcgac aggcgccgga cctctggtga ggggagggat aagtgaggcg
     3241 tcagtttctt tggtcggttt tatgtaccta tcttcttaag tagctgaagc tccggttttg
     3301 aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat
     3361 gtaatcattt gggtcaatat gtaattttca gtgttagact tgtaaattgt ccgctaaatt
     3421 ctggccgttt ttggcttttt tgttagacaa catgggtaaa aagcctgaac tcaccgcgac
     3481 gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc
     3541 ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg
     3601 ggtaaatagc tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc
     3661 cgccgcgctc ccgattccgg aagtgcttga cattggggag ttcagcgaga gcctgaccta
     3721 ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc
     3781 cgctgttctg cagccggtcg cggaggcaat ggatgccatc gctgccgccg atcttagcca
     3841 gacgagcggg ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga
     3901 tttcatatgc gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac
     3961 cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc
     4021 cgaagtccgg cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg
     4081 ccgcataaca gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt
     4141 cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt
     4201 cgagcggagg catccggagc ttgcaggatc gccccggctc cgggcgtata tgctccgcat
     4261 tggtcttgac caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc
     4321 gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat
     4381 cgcccgcaga agcgccgccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg
     4441 aaaccgacgc cccagcactc gtccgagggc aaaggaataa gctagtatgt aagcctagtc
     4501 ttagataata aaatcgctat ccatcgaaga tggatgtgtg ttggtttttt gtgtgtgtaa
     4561 cgctaggcgc gcctggtgta ccgagaacga tcctctcagt gcgagtctcg acgatccata
     4621 tcgttgcttg gcagtcagcc agtcggaatc cagcttggga cccaggaagt ccaatcgtca
     4681 gatattgtac tcaagcctgg tcacggcagc gtaccgatct gtttaaacct agatattgat
     4741 agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc aaacatctat caagagacag
     4801 gatcagcagg aggctttcgc atgagtattc aacatttccg tgtcgccctt attccctttt
     4861 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     4921 ctgaagatca gttgggtgcg cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     4981 tccttgagag ttttcgcccc gaagaacgct ttccaatgat gagcactttt aaagttctgc
     5041 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     5101 actattctca gaatgacttg gttgagtatt caccagtcac agaaaagcat cttacggatg
     5161 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     5221 acttacttct gacaacgatt ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5281 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5341 acgagcgtga caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa ctattaactg
     5401 gcgaactact tactctagct tcccggcaac agttgataga ctggatggag gcggataaag
     5461 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5521 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5581 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5641 agatcgctga gataggtgcc tcactgatta agcattggta accgattcta ggtgcattgg
     5701 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
     5761 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
     5821 ttatccttaa gatcccgaat cgtttaaact cgactctggc tctatcgaat ctccgtcgtt
     5881 tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc
     5941 tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg acatcttgga ccattagctc
     6001 cacaggtatc ttcttccctc tagtggtcat aacagcagct tcagctacct ctcaattcaa
     6061 aaaacccctc aagacccgtt tagaggcccc aaggggttat gctatcaatc gttgcgttac
     6121 acacacaaaa aaccaacaca catccatctt cgatggatag cgattttatt atctaactgc
     6181 tgatcgagtg tagccagatc tagtaatcaa ttacggggtc attagttcat agcccatata
     6241 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     6301 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     6361 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     6421 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     6481 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     6541 tcgctattac catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     6601 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
     6661 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
     6721 gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca
     6781 gatctttgtc gatcctacca tccactcgac acacccgcca gcggccgc
//
