LOCUS       sequence-301673-        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-m4252903
FEATURES             Location/Qualifiers
     source          1..6828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      48..94
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    99..170
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(322..458)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(471..1262,1263..1331))
                     /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     1094..1113
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1597..1645
                     /note="synthetic polyadenylation signal"
     CDS             complement(1672..2700)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(2704..3913)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(2848..2867)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      4413..5001
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4902..4921
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5227..5298
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5343..5479)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(5509..5555)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5670..5791
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5707..5726)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5761..5780
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(5860..5901)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(5902..5922)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(6222..6424)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6251..6271)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6426..6729
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    join(6810..6828,1..30)
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 gaagatggat gtgtgttggt tttttgtgtg tgtaacgcaa cgattgatag cataacccct
       61 tggggcctct aaacgggtct tgaggggttt tttgaattga gaggtagctg aagctgctgt
      121 tatgaccact agagggaaga agatacctgt ggagctaatg gtccaagatg tcgggagccg
      181 cgatcgctgc caaaaaggta agctgacgat agctgacgag attcgatgcc cgacgagcaa
      241 atgagcgcca cggctgttcg cgtaagctcg aaacgacgga gattcgatag agccagagtc
      301 gagtttaaac gattcgggat cttaaggata aatttctggt aaggaggaca cgtatggaag
      361 tgggcaagtt ggggaagccg tatccgttgc tgaatctggc atatgtggga gtataagacg
      421 cgcagcgtcg catcaggcat ttttttctgc gccaatgcac ctagaatcgg ttaccaatgc
      481 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      541 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      601 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      661 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctatcaac
      721 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttacgcaa ggttgttgcc
      781 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      841 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      901 ttcggtcctc caatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      961 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     1021 gaatactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     1081 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     1141 aagcgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     1201 taacccactc gcgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     1261 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1321 tgaatactca tgcgaaagcc tcctgctgat cctgtctctt gatagatgtt tgcaaaagct
     1381 aggactcgat tatacgttga ccgatcagac tatcaatatc taggtttaaa cagatcggta
     1441 cgctgccgtg accaggcttg agtacaatat ctgacgattg gacttcctgg gtcccaagct
     1501 ggattccgac tggctgactg ccaagcaacg atatggatcg tcgagactcg cactgagagg
     1561 atcgttctcg gtacaccagg cgcgcctagc gttacacaca caaaaaacca acacacatcc
     1621 atcttcgatg gatagcgatt ttattatcta agactaggct tacatactag cttattcctt
     1681 tgccctcgga cgagtgctgg ggcgtcggtt tccactatcg gcgagtactt ctacacagcc
     1741 atcggtccag acggcggcgc ttctgcgggc gatttgtgta cgcccgacag tcccggctcc
     1801 ggatcggacg attgcgtcgc atcgaccctg cgcccaagct gcatcatcga aattgccgtc
     1861 aaccaagctc tgatagagtt ggtcaagacc aatgcggagc atatacgccc ggagccgggg
     1921 cgatcctgca agctccggat gcctccgctc gaagtagcgc gtctgctgct ccatacaagc
     1981 caaccacggc ctccagaaga agatgttggc gacctcgtat tgggaatccc cgaacatcgc
     2041 ctcgctccag tcaatgaccg ctgttatgcg gccattgtcc gtcaggacat tgttggagcc
     2101 gaaatccgcg tgcacgaggt gccggacttc ggggcagtcc tcggcccaaa gcatcagctc
     2161 atcgagagcc tgcgcgacgg acgcactgac ggtgtcgtcc atcacagttt gccagtgata
     2221 cacatgggga tcagcaatcg cgcatatgaa atcacgccat gtagtgtatt gaccgattcc
     2281 ttgcggtccg aatgggccga acccgctcgt ctggctaaga tcggcggcag cgatggcatc
     2341 cattgcctcc gcgaccggct gcagaacagc gggcagttcg gtttcaggca ggtcttgcaa
     2401 cgtgacaccc tgtgcacggc gggagatgca ataggtcagg ctctcgctga actccccaat
     2461 gtcaagcact tccggaatcg ggagcgcggc ggatgcaaag tgccgataaa cataacgatc
     2521 tttgtagaaa ccatcggcgc agctatttac ccgcaggaca tatccacgcc ctcctacatc
     2581 gaagctgaaa gcacgagatt cttcgccctc cgagagctgc atcaggtcgg agacgctgtc
     2641 gaacttttcg atcagaaact tctcgacaga cgtcgcggtg agttcaggct ttttacccat
     2701 gttgtctaac aaaaaagcca aaaacggcca gaatttagcg gacaatttac aagtctaaca
     2761 ctgaaaatta catattgacc caaatgatta catttcaaaa ggtgcctaaa aaacttcaca
     2821 aaacacactc gccaaccccg agcgcatagt tcaaaaccgg agcttcagct acttaagaag
     2881 ataggtacat aaaaccgacc aaagaaactg acgcctcact tatccctccc ctcaccagag
     2941 gtccggcgcc tgtcgattca ggagagccta ccctaggccc gaaccctgcg tcctgcgacg
     3001 gagaaaagcc taccgcacac ctaccggctg gtggccccac cctgcattat aagccaacag
     3061 aacgggtgac gtcacgacac gacgagggcg cgcgctccca aaggtacggg tgcactgccc
     3121 aacggcaccg ccataactgc cgcccccgca acagacgaca aaccgagttc tccagtcagt
     3181 gacaaacttc acgtcagggt ccccagatgg tgcccagccc atctcacccg aataagagct
     3241 ttcccgcatt agcgaagggc tcaagacctt gggttcttgc cgcccaccat gccccccacc
     3301 ttgtttcaac gacctcacag cccgcctcac aagcgtcttc cattcaagac tcgggaacag
     3361 ccgccatttt gctgcgctcc ccccaacccc cagttcaggg caaccttgct cgcggaccca
     3421 gactacagcc cttggcggtc tctccacacg cttccgtccc accgagcggc ccggcggcca
     3481 cgaaagcccc ggccagccca gcagcccgct actcaccaag tgacgatcac agcgatccac
     3541 aaacaagaac cgcgacccaa atcccggctg cgacggaact agctgtgcca cacccggcgc
     3601 gtccttatat aatcatcggc gttcaccgcc ccacggagat ccctccgcag aatcgccgag
     3661 aagggactac ttttcctcgc ctgttccgct ctctggaaag aaaaccagtg ccctagagtc
     3721 acccaagtcc cgtcctaaaa tgtccttctg ctgatactgg ggttctaagg ccgagtctta
     3781 tgagcagcgg gccgctgtcc tgagcgtccg ggcggaagga tcaggacgct cctgcgccct
     3841 tcgtctgacg tggcagcgct cgccgtgacg aggggggcgc ccgcgggagg cgccaaaacc
     3901 cggcgcggag gccaggcgcg ccccgatttc gatattcctc ggatctcgga tagatcggga
     3961 ttgaggatgt aggttgaact tctctcgggt gatcccgatc tcggatattc gggattggag
     4021 taatacctgg cgataggcgc tgccaagcaa tagccaaggc gcaaggccgg ccaagagact
     4081 tgcgatagtt cgttcgagag gtagcctgca agatggatcg gacgaaaggt ttgaagattt
     4141 tccgacgagg gccttggaga gttcggaaat ttaaatgcaa ctatggatga acgaaataga
     4201 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     4261 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     4321 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4381 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4441 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4501 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     4561 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4621 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4681 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4741 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4801 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4861 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4921 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4981 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5041 tgctggcctt ttgctcacat gatttaaatg aaactcgtaa tcggagaacg ggtgcaatag
     5101 caaggaccag atcgaacttg actgacgcaa ttctgagcgt agtcgaaacc cagacaagtc
     5161 gatctcgcca agtagctgac ttgactggtt aattaagttt cgatagccca aggtaaccaa
     5221 tgagattagg gagggaagaa agtttgatga actacttctg acccctagtg gtgtccagaa
     5281 aagaccatta aaggaatgtc ttgatcgaaa tcgagagatc gcctgcagga tagctgacga
     5341 ccttaaggat aaatttctgg taaggaggac acgtatggaa gtgggcaagt tggggaagcc
     5401 gtatccgttg ctgaatctgg catatgtggg agtataagac gcgcagcgtc gcatcaggca
     5461 tttttttctg cgccaatgcc ggtagatagc aatccggccc gaggggcaca aaaaacccct
     5521 caagacccgt ttagaggccc caaggggtta tgctacgata ccgatagaga tgggccaata
     5581 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     5641 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     5701 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5761 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tgagctgaag gtacgctgta
     5821 tctcagtcag tcaagctagc agaggcctgc ggatccttag gtagaatcta agccgagaag
     5881 tggattgggt ataggtttcc ccgactgaaa gtaaaggttt tcgacaagaa gatcaggcac
     5941 acgggagctg tttagatttt tcacgcggga ataaacgtag aacgagggct tcacgagact
     6001 aacgttgaca atgttacaac aataagcgca cagccggagg gccgtcagta tcgctagggt
     6061 cactaaaagg aagaccacaa atgccaggaa cagcaacact gaattaacga tgagagtgcc
     6121 tgtctcttca gatacaaagc tgtacatggt ggcggccgct ggcgggtgtg tcgagtggat
     6181 ggtaggatcg acaaagatct gatctgacgg ttcactaaac cagctctgct tatatagacc
     6241 tcccaccgta cacgcctacc gcccatttgc gtcaatgggg cggagttgtt acgacatttt
     6301 ggaaagtccc gttgattttg gtgccaaaac aaactcccat tgacgtcaat ggggtggaga
     6361 cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc
     6421 atcagcatgg taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat
     6481 aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg
     6541 ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac
     6601 tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt
     6661 attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt
     6721 tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat tgattactag
     6781 atctggctac actcgatcag cagttagata ataaaatcgc tatccatc
//
