LOCUS       sequence-301672-        7176 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 7176)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7176)
  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-m4252968
FEATURES             Location/Qualifiers
     source          1..7176
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    47..168
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(58..77)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     112..131
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      283..329
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      359..495
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    540..611
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(837..1425)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(917..936)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1925..3134
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     2971..2990
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3138..4166
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4193..4241
                     /note="synthetic polyadenylation signal"
     CDS             join(4507..4575,4576..5367)
                     /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(4725..4744)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5380..5516
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5668..5739
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5744..5790)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5808..5856
                     /note="synthetic polyadenylation signal"
     enhancer        5937..6240
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6242..6444
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6395..6415
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             6744..6764
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             7113..7130
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 agcttgactg actgagatac agcgtacctt cagctcacag acatgataag atacattgat
       61 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
      121 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
      181 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
      241 aacctctaca aatgtggtat tggcccatct ctatcggtat cgtagcataa ccccttgggg
      301 cctctaaacg ggtcttgagg ggttttttgt gcccctcggg ccggattgct atctaccggc
      361 attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca
      421 gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca
      481 gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat ctctcgattt cgatcaagac
      541 attcctttaa tggtcttttc tggacaccac taggggtcag aagtagttca tcaaactttc
      601 ttccctccct aatctcattg gttaccttgg gctatcgaaa cttaattaac cagtcaagtc
      661 agctacttgg cgagatcgac ttgtctgggt ttcgactacg ctcagaattg cgtcagtcaa
      721 gttcgatctg gtccttgcta ttgcacccgt tctccgatta cgagtttcat ttaaatcatg
      781 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
      841 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      901 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      961 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     1021 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     1081 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     1141 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     1201 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     1261 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     1321 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     1381 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     1441 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     1501 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     1561 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     1621 cctatctcag cgatctgtct atttcgttca tccatagttg catttaaatt tccgaactct
     1681 ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga tccatcttgc aggctacctc
     1741 tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc ttggctattg cttggcagcg
     1801 cctatcgcca ggtattactc caatcccgaa tatccgagat cgggatcacc cgagagaagt
     1861 tcaacctaca tcctcaatcc cgatctatcc gagatccgag gaatatcgaa atcggggcgc
     1921 gcctggcctc cgcgccgggt tttggcgcct cccgcgggcg cccccctcgt cacggcgagc
     1981 gctgccacgt cagacgaagg gcgcaggagc gtcctgatcc ttccgcccgg acgctcagga
     2041 cagcggcccg ctgctcataa gactcggcct tagaacccca gtatcagcag aaggacattt
     2101 taggacggga cttgggtgac tctagggcac tggttttctt tccagagagc ggaacaggcg
     2161 aggaaaagta gtcccttctc ggcgattctg cggagggatc tccgtggggc ggtgaacgcc
     2221 gatgattata taaggacgcg ccgggtgtgg cacagctagt tccgtcgcag ccgggatttg
     2281 ggtcgcggtt cttgtttgtg gatcgctgtg atcgtcactt ggtgagtagc gggctgctgg
     2341 gctggccggg gctttcgtgg ccgccgggcc gctcggtggg acggaagcgt gtggagagac
     2401 cgccaagggc tgtagtctgg gtccgcgagc aaggttgccc tgaactgggg gttgggggga
     2461 gcgcagcaaa atggcggctg ttcccgagtc ttgaatggaa gacgcttgtg aggcgggctg
     2521 tgaggtcgtt gaaacaaggt ggggggcatg gtgggcggca agaacccaag gtcttgagcc
     2581 cttcgctaat gcgggaaagc tcttattcgg gtgagatggg ctgggcacca tctggggacc
     2641 ctgacgtgaa gtttgtcact gactggagaa ctcggtttgt cgtctgttgc gggggcggca
     2701 gttatggcgg tgccgttggg cagtgcaccc gtacctttgg gagcgcgcgc cctcgtcgtg
     2761 tcgtgacgtc acccgttctg ttggcttata atgcagggtg gggccaccag ccggtaggtg
     2821 tgcggtaggc ttttctccgt cgcaggacgc agggttcggg cctagggtag gctctcctga
     2881 atcgacaggc gccggacctc tggtgagggg agggataagt gaggcgtcag tttctttggt
     2941 cggttttatg tacctatctt cttaagtagc tgaagctccg gttttgaact atgcgctcgg
     3001 ggttggcgag tgtgttttgt gaagtttttt aggcaccttt tgaaatgtaa tcatttgggt
     3061 caatatgtaa ttttcagtgt tagacttgta aattgtccgc taaattctgg ccgtttttgg
     3121 cttttttgtt agacaacatg ggtaaaaagc ctgaactcac cgcgacgtct gtcgagaagt
     3181 ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat
     3241 ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg
     3301 ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatccgcc gcgctcccga
     3361 ttccggaagt gcttgacatt ggggagttca gcgagagcct gacctattgc atctcccgcc
     3421 gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc
     3481 cggtcgcgga ggcaatggat gccatcgctg ccgccgatct tagccagacg agcgggttcg
     3541 gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga
     3601 ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg
     3661 tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc
     3721 tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg
     3781 tcattgactg gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct
     3841 tctggaggcc gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc
     3901 cggagcttgc aggatcgccc cggctccggg cgtatatgct ccgcattggt cttgaccaac
     3961 tctatcagag cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg
     4021 acgcaatcgt ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg
     4081 ccgccgtctg gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca
     4141 gcactcgtcc gagggcaaag gaataagcta gtatgtaagc ctagtcttag ataataaaat
     4201 cgctatccat cgaagatgga tgtgtgttgg ttttttgtgt gtgtaacgct aggcgcgcct
     4261 ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga tccatatcgt tgcttggcag
     4321 tcagccagtc ggaatccagc ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa
     4381 gcctggtcac ggcagcgtac cgatctgttt aaacctagat attgatagtc tgatcggtca
     4441 acgtataatc gagtcctagc ttttgcaaac atctatcaag agacaggatc agcaggaggc
     4501 tttcgcatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     4561 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     4621 ggtgcgcgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     4681 cgccccgaag aacgctttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     4741 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     4801 gacttggttg agtattcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     4861 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     4921 acgattggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     4981 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5041 acgatgcctg tagcaatggc aacaaccttg cgtaaactat taactggcga actacttact
     5101 ctagcttccc ggcaacagtt gatagactgg atggaggcgg ataaagttgc aggaccactt
     5161 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     5221 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     5281 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     5341 ggtgcctcac tgattaagca ttggtaaccg attctaggtg cattggcgca gaaaaaaatg
     5401 cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg
     5461 cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaagatc
     5521 ccgaatcgtt taaactcgac tctggctcta tcgaatctcc gtcgtttcga gcttacgcga
     5581 acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac
     5641 ctttttggca gcgatcgcgg ctcccgacat cttggaccat tagctccaca ggtatcttct
     5701 tccctctagt ggtcataaca gcagcttcag ctacctctca attcaaaaaa cccctcaaga
     5761 cccgtttaga ggccccaagg ggttatgcta tcaatcgttg cgttacacac acaaaaaacc
     5821 aacacacatc catcttcgat ggatagcgat tttattatct aactgctgat cgagtgtagc
     5881 cagatctagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     5941 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     6001 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     6061 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     6121 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     6181 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     6241 ctgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     6301 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     6361 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     6421 tgggaggtct atataagcag agctggttta gtgaaccgtc agatcagatc tttgtcgatc
     6481 ctaccatcca ctcgacacac ccgccagcgg ccgccaccat gtacagcttt gtatctgaag
     6541 agacaggcac tctcatcgtt aattcagtgt tgctgttcct ggcatttgtg gtcttccttt
     6601 tagtgaccct agcgatactg acggccctcc ggctgtgcgc ttattgttgt aacattgtca
     6661 acgttagtct cgtgaagccc tcgttctacg tttattcccg cgtgaaaaat ctaaacagct
     6721 cccgtgtgcc tgatcttctt gtcgaaaacc tttactttca gtcgatgagt gacaaaatca
     6781 ttcatctaac cgatgactcc tttgatacag acgtgttgaa ggccgatggt gcgatactgg
     6841 tagatttctg ggctgaatgg tgtggcccgt gcaagatgat cgcacccatt ctggacgaga
     6901 tcgccgacga gtatcaagga aaattaactg tcgctaagct taatatagat cagaacccag
     6961 ggacggcacc taaatacggc atcaggggaa tacctaccct cctgctcttt aagaatgggg
     7021 aagttgccgc gacaaaagtg ggagccctgt ctaagggcca gctaaaagag ttcttggacg
     7081 ctaacctggc aggttcaggc tcggggcaca tgcaccatca ccatcaccat agcagcgggc
     7141 tcgtgccacg cggctaagga tccgcaggcc tctgct
//
