LOCUS       sequence-307244-        6368 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 6368)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6368)
  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-m4252825
FEATURES             Location/Qualifiers
     source          1..6368
                     /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..277
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    330..451
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(341..360)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     395..414
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      566..612
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      642..778
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    823..894
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1120..1708)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1200..1219)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2208..3417
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3254..3273
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3421..4020
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3421..3440)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3917..3937
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4047..4095
                     /note="synthetic polyadenylation signal"
     CDS             join(4361..4429,4430..5221)
                     /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(4579..4598)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5234..5370
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5522..5593
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5598..5644)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5662..5710
                     /note="synthetic polyadenylation signal"
     enhancer        5791..6094
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6096..6298
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6249..6269
                     /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 tacccctatg acgtaccaga ttatgcgtaa gctagcttga ctgactgaga
      301 tacagcgtac cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac
      361 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
      421 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
      481 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
      541 tattggccca tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg
      601 aggggttttt tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa
      661 tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac
      721 ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg
      781 tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt
      841 ttctggacac cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca
      901 ttggttacct tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc
      961 gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg
     1021 ctattgcacc cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca
     1081 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1141 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1201 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1261 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1321 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1381 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1441 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1501 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1561 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1621 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1681 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     1741 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     1801 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     1861 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     1921 tctatttcgt tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga
     1981 aaatcttcaa acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca
     2041 agtctcttgg ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta
     2101 ctccaatccc gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa
     2161 tcccgatcta tccgagatcc gaggaatatc gaaatcgggg cgcgcctggc ctccgcgccg
     2221 ggttttggcg cctcccgcgg gcgcccccct cgtcacggcg agcgctgcca cgtcagacga
     2281 agggcgcagg agcgtcctga tccttccgcc cggacgctca ggacagcggc ccgctgctca
     2341 taagactcgg ccttagaacc ccagtatcag cagaaggaca ttttaggacg ggacttgggt
     2401 gactctaggg cactggtttt ctttccagag agcggaacag gcgaggaaaa gtagtccctt
     2461 ctcggcgatt ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt atataaggac
     2521 gcgccgggtg tggcacagct agttccgtcg cagccgggat ttgggtcgcg gttcttgttt
     2581 gtggatcgct gtgatcgtca cttggtgagt agcgggctgc tgggctggcc ggggctttcg
     2641 tggccgccgg gccgctcggt gggacggaag cgtgtggaga gaccgccaag ggctgtagtc
     2701 tgggtccgcg agcaaggttg ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg
     2761 ctgttcccga gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa
     2821 ggtggggggc atggtgggcg gcaagaaccc aaggtcttga gcccttcgct aatgcgggaa
     2881 agctcttatt cgggtgagat gggctgggca ccatctgggg accctgacgt gaagtttgtc
     2941 actgactgga gaactcggtt tgtcgtctgt tgcgggggcg gcagttatgg cggtgccgtt
     3001 gggcagtgca cccgtacctt tgggagcgcg cgccctcgtc gtgtcgtgac gtcacccgtt
     3061 ctgttggctt ataatgcagg gtggggccac cagccggtag gtgtgcggta ggcttttctc
     3121 cgtcgcagga cgcagggttc gggcctaggg taggctctcc tgaatcgaca ggcgccggac
     3181 ctctggtgag gggagggata agtgaggcgt cagtttcttt ggtcggtttt atgtacctat
     3241 cttcttaagt agctgaagct ccggttttga actatgcgct cggggttggc gagtgtgttt
     3301 tgtgaagttt tttaggcacc ttttgaaatg taatcatttg ggtcaatatg taattttcag
     3361 tgttagactt gtaaattgtc cgctaaattc tggccgtttt tggctttttt gttagacaac
     3421 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
     3481 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
     3541 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     3601 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     3661 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     3721 tcccggctgg ccgcgcagca acagatggag ggcctcctgg cgccgcaccg gcccaaggag
     3781 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     3841 agcgccgtcg tgctccccgg agtggaggcc gccgagcgcg ccggggtgcc cgccttcctg
     3901 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     3961 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctag
     4021 gctagtatgt aagcctagtc ttagataata aaatcgctat ccatcgaaga tggatgtgtg
     4081 ttggtttttt gtgtgtgtaa cgctaggcgc gcctggtgta ccgagaacga tcctctcagt
     4141 gcgagtctcg acgatccata tcgttgcttg gcagtcagcc agtcggaatc cagcttggga
     4201 cccaggaagt ccaatcgtca gatattgtac tcaagcctgg tcacggcagc gtaccgatct
     4261 gtttaaacct agatattgat agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc
     4321 aaacatctat caagagacag gatcagcagg aggctttcgc atgagtattc aacatttccg
     4381 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     4441 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt acatcgaact
     4501 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgct ttccaatgat
     4561 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     4621 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtatt caccagtcac
     4681 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     4741 gagtgataac actgcggcca acttacttct gacaacgatt ggaggaccga aggagctaac
     4801 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     4861 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     4921 cttgcgtaaa ctattaactg gcgaactact tactctagct tcccggcaac agttgataga
     4981 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     5041 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     5101 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     5161 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     5221 accgattcta ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata
     5281 ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc acttccatac
     5341 gtgtcctcct taccagaaat ttatccttaa ggtcccgaat cgtttaaact cgactctggc
     5401 tctatcgaat ctccgtcgtt tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc
     5461 gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg
     5521 acatcttgga ccattagctc cacaggtatc ttcttccctc tagtggtcat aacagcagct
     5581 tcagctacct ctcaattcaa aaaacccctc aagacccgtt tagaggcccc aaggggttat
     5641 gctatcaatc gttgcgttac acacacaaaa aaccaacaca catccatctt cgatggatag
     5701 cgattttatt atctaactgc tgatcgagtg tagccagatc tagtaatcaa ttacggggtc
     5761 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     5821 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     5881 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     5941 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     6001 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     6061 gtacatctac gtattagtca tcgctattac catgctgatg cggttttggc agtacatcaa
     6121 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     6181 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     6241 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
     6301 tttagtgaac cgtcagatca gatctttgtc gatcctacca tccactcgac acacccgcca
     6361 gcggccgc
//
