LOCUS       sequence-300709-        6804 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 6804)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6804)
  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-m4252912
FEATURES             Location/Qualifiers
     source          1..6804
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(344..1553)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(488..507)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      2053..2641
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2542..2561
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2867..2938
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2983..3119)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(3149..3195)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3310..3431
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3347..3366)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3401..3420
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(3500..3517)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(3518..3538)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(3838..4040)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3867..3887)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4042..4345
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    4426..4474
                     /note="synthetic polyadenylation signal"
     terminator      4492..4538
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    4543..4614
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4766..4902)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(4915..5706,5707..5775))
                     /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     5538..5557
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    6041..6089
                     /note="synthetic polyadenylation signal"
     CDS             complement(join(6116..6804,1..340))
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
ORIGIN
        1 gcagaacagc gggcagttcg gtttcaggca ggtcttgcaa cgtgacaccc tgtgcacggc
       61 gggagatgca ataggtcagg ctctcgctga actccccaat gtcaagcact tccggaatcg
      121 ggagcgcggc ggatgcaaag tgccgataaa cataacgatc tttgtagaaa ccatcggcgc
      181 agctatttac ccgcaggaca tatccacgcc ctcctacatc gaagctgaaa gcacgagatt
      241 cttcgccctc cgagagctgc atcaggtcgg agacgctgtc gaacttttcg atcagaaact
      301 tctcgacaga cgtcgcggtg agttcaggct ttttacccat gttgtctaac aaaaaagcca
      361 aaaacggcca gaatttagcg gacaatttac aagtctaaca ctgaaaatta catattgacc
      421 caaatgatta catttcaaaa ggtgcctaaa aaacttcaca aaacacactc gccaaccccg
      481 agcgcatagt tcaaaaccgg agcttcagct acttaagaag ataggtacat aaaaccgacc
      541 aaagaaactg acgcctcact tatccctccc ctcaccagag gtccggcgcc tgtcgattca
      601 ggagagccta ccctaggccc gaaccctgcg tcctgcgacg gagaaaagcc taccgcacac
      661 ctaccggctg gtggccccac cctgcattat aagccaacag aacgggtgac gtcacgacac
      721 gacgagggcg cgcgctccca aaggtacggg tgcactgccc aacggcaccg ccataactgc
      781 cgcccccgca acagacgaca aaccgagttc tccagtcagt gacaaacttc acgtcagggt
      841 ccccagatgg tgcccagccc atctcacccg aataagagct ttcccgcatt agcgaagggc
      901 tcaagacctt gggttcttgc cgcccaccat gccccccacc ttgtttcaac gacctcacag
      961 cccgcctcac aagcgtcttc cattcaagac tcgggaacag ccgccatttt gctgcgctcc
     1021 ccccaacccc cagttcaggg caaccttgct cgcggaccca gactacagcc cttggcggtc
     1081 tctccacacg cttccgtccc accgagcggc ccggcggcca cgaaagcccc ggccagccca
     1141 gcagcccgct actcaccaag tgacgatcac agcgatccac aaacaagaac cgcgacccaa
     1201 atcccggctg cgacggaact agctgtgcca cacccggcgc gtccttatat aatcatcggc
     1261 gttcaccgcc ccacggagat ccctccgcag aatcgccgag aagggactac ttttcctcgc
     1321 ctgttccgct ctctggaaag aaaaccagtg ccctagagtc acccaagtcc cgtcctaaaa
     1381 tgtccttctg ctgatactgg ggttctaagg ccgagtctta tgagcagcgg gccgctgtcc
     1441 tgagcgtccg ggcggaagga tcaggacgct cctgcgccct tcgtctgacg tggcagcgct
     1501 cgccgtgacg aggggggcgc ccgcgggagg cgccaaaacc cggcgcggag gccaggcgcg
     1561 ccccgatttc gatattcctc ggatctcgga tagatcggga ttgaggatgt aggttgaact
     1621 tctctcgggt gatcccgatc tcggatattc gggattggag taatacctgg cgataggcgc
     1681 tgccaagcaa tagccaaggc gcaaggccgg ccaagagact tgcgatagtt cgttcgagag
     1741 gtagcctgca agatggatcg gacgaaaggt ttgaagattt tccgacgagg gccttggaga
     1801 gttcggaaat ttaaatgcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     1861 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     1921 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     1981 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     2041 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2101 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2161 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     2221 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2281 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     2341 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     2401 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     2461 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     2521 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     2581 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     2641 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     2701 gatttaaatg aaactcgtaa tcggagaacg ggtgcaatag caaggaccag atcgaacttg
     2761 actgacgcaa ttctgagcgt agtcgaaacc cagacaagtc gatctcgcca agtagctgac
     2821 ttgactggtt aattaagttt cgatagccca aggtaaccaa tgagattagg gagggaagaa
     2881 agtttgatga actacttctg acccctagtg gtgtccagaa aagaccatta aaggaatgtc
     2941 ttgatcgaaa tcgagagatc gcctgcagga tagctgacga ccttaaggat aaatttctgg
     3001 taaggaggac acgtatggaa gtgggcaagt tggggaagcc gtatccgttg ctgaatctgg
     3061 catatgtggg agtataagac gcgcagcgtc gcatcaggca tttttttctg cgccaatgcc
     3121 ggtagatagc aatccggccc gaggggcaca aaaaacccct caagacccgt ttagaggccc
     3181 caaggggtta tgctacgata ccgatagaga tgggccaata ccacatttgt agaggtttta
     3241 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     3301 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3361 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3421 aatgtatctt atcatgtctg tgagctgaag gtacgctgta tctcagtcag tcaagctagc
     3481 agaggcctgc ggatccttaa tggtgatggt gatggtgcga ctgaaagtaa aggttttcga
     3541 ccagaagatc aggaactcta gaagaattca gatttttaac acgagagtaa acgtaaaaag
     3601 aaggttttac aagactcacg ttaacaatat tgcagcagta cgcacacaat cgaagcgcag
     3661 taaggatggc tagtgtaact agcaagaata ccacgaaagc aagaaaaaga agtacgctat
     3721 taactattaa cgtacctgtc tcttccgaaa cgaatgagta catggtggcg gccgctggcg
     3781 ggtgtgtcga gtggatggta ggatcgacaa agatctgatc tgacggttca ctaaaccagc
     3841 tctgcttata tagacctccc accgtacacg cctaccgccc atttgcgtca atggggcgga
     3901 gttgttacga cattttggaa agtcccgttg attttggtgc caaaacaaac tcccattgac
     3961 gtcaatgggg tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattgat
     4021 gtactgccaa aaccgcatca gcatggtaat agcgatgact aatacgtaga tgtactgcca
     4081 agtaggaaag tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc
     4141 attgacgtca atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc
     4201 agtttaccgt aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat
     4261 gggaacatac gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg
     4321 gccatttacc gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc
     4381 cgtaattgat tactagatct ggctacactc gatcagcagt tagataataa aatcgctatc
     4441 catcgaagat ggatgtgtgt tggttttttg tgtgtgtaac gcaacgattg atagcataac
     4501 cccttggggc ctctaaacgg gtcttgaggg gttttttgaa ttgagaggta gctgaagctg
     4561 ctgttatgac cactagaggg aagaagatac ctgtggagct aatggtccaa gatgtcggga
     4621 gccgcgatcg ctgccaaaaa ggtaagctga cgatagctga cgagattcga tgcccgacga
     4681 gcaaatgagc gccacggctg ttcgcgtaag ctcgaaacga cggagattcg atagagccag
     4741 agtcgagttt aaacgattcg ggatcttaag gataaatttc tggtaaggag gacacgtatg
     4801 gaagtgggca agttggggaa gccgtatccg ttgctgaatc tggcatatgt gggagtataa
     4861 gacgcgcagc gtcgcatcag gcattttttt ctgcgccaat gcacctagaa tcggttacca
     4921 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4981 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5041 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5101 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5161 caactgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttac gcaaggttgt
     5221 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5281 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5341 ctccttcggt cctccaatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5401 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5461 tggtgaatac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5521 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5581 tggaaagcgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5641 gatgtaaccc actcgcgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5701 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5761 atgttgaata ctcatgcgaa agcctcctgc tgatcctgtc tcttgataga tgtttgcaaa
     5821 agctaggact cgattatacg ttgaccgatc agactatcaa tatctaggtt taaacagatc
     5881 ggtacgctgc cgtgaccagg cttgagtaca atatctgacg attggacttc ctgggtccca
     5941 agctggattc cgactggctg actgccaagc aacgatatgg atcgtcgaga ctcgcactga
     6001 gaggatcgtt ctcggtacac caggcgcgcc tagcgttaca cacacaaaaa accaacacac
     6061 atccatcttc gatggatagc gattttatta tctaagacta ggcttacata ctagcttatt
     6121 cctttgccct cggacgagtg ctggggcgtc ggtttccact atcggcgagt acttctacac
     6181 agccatcggt ccagacggcg gcgcttctgc gggcgatttg tgtacgcccg acagtcccgg
     6241 ctccggatcg gacgattgcg tcgcatcgac cctgcgccca agctgcatca tcgaaattgc
     6301 cgtcaaccaa gctctgatag agttggtcaa gaccaatgcg gagcatatac gcccggagcc
     6361 ggggcgatcc tgcaagctcc ggatgcctcc gctcgaagta gcgcgtctgc tgctccatac
     6421 aagccaacca cggcctccag aagaagatgt tggcgacctc gtattgggaa tccccgaaca
     6481 tcgcctcgct ccagtcaatg accgctgtta tgcggccatt gtccgtcagg acattgttgg
     6541 agccgaaatc cgcgtgcacg aggtgccgga cttcggggca gtcctcggcc caaagcatca
     6601 gctcatcgag agcctgcgcg acggacgcac tgacggtgtc gtccatcaca gtttgccagt
     6661 gatacacatg gggatcagca atcgcgcata tgaaatcacg ccatgtagtg tattgaccga
     6721 ttccttgcgg tccgaatggg ccgaacccgc tcgtctggct aagatcggcg gcagcgatgg
     6781 catccattgc ctccgcgacc ggct
//
