LOCUS       sequence-300230-        6410 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 6410)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6410)
  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-m4133579
FEATURES             Location/Qualifiers
     source          1..6410
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    65..136
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(141..187)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    205..253
                     /note="synthetic polyadenylation signal"
     enhancer        334..637
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        639..841
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     792..812
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1141..1161
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1165..1230
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1283..1404
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1294..1313)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1348..1367
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1519..1565
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1595..1731
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1776..1847
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2073..2661)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2153..2172)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3161..4370
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4207..4226
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4374..4973
                     /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(4374..4393)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4870..4890
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5000..5048
                     /note="synthetic polyadenylation signal"
     CDS             join(5314..5382,5383..6174)
                     /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(5532..5551)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6187..6323
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg atcgcggctc
       61 ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt cataacagca
      121 gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt
      181 tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat cttcgatgga
      241 tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat caattacggg
      301 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      361 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      421 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      481 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      541 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      601 gcagtacatc tacgtattag tcatcgctat taccatgctg atgcggtttt ggcagtacat
      661 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      721 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
      781 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      841 tggtttagtg aaccgtcaga tcagatcttt gtcgatccta ccatccactc gacacacccg
      901 ccagcggccg ccaccatgta ctcattcgtt tcggaagaga caggtacgtt aatagttaat
      961 agcgtacttc tttttcttgc tttcgtggta ttcttgctag ttacactagc catccttact
     1021 gcgcttcgat tgtgtgcgta ctgctgcaat attgttaacg tgagtcttgt aaaaccttct
     1081 ttttacgttt actctcgtgt taaaaatctg aattcttcta gagttcctga tcttctggtc
     1141 gaaaaccttt actttcagtc gatggattat aaggaccacg atggagacta caaagatcat
     1201 gacattgatt acaaggacga cgatgacaaa taagctagct tgactgactg agatacagcg
     1261 taccttcagc tcacagacat gataagatac attgatgagt ttggacaaac cacaactaga
     1321 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
     1381 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
     1441 cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtattggc
     1501 ccatctctat cggtatcgta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     1561 ttttgtgccc ctcgggccgg attgctatct accggcattg gcgcagaaaa aaatgcctga
     1621 tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga tacggcttcc
     1681 ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta aggtcgtcag
     1741 ctatcctgca ggcgatctct cgatttcgat caagacattc ctttaatggt cttttctgga
     1801 caccactagg ggtcagaagt agttcatcaa actttcttcc ctccctaatc tcattggtta
     1861 ccttgggcta tcgaaactta attaaccagt caagtcagct acttggcgag atcgacttgt
     1921 ctgggtttcg actacgctca gaattgcgtc agtcaagttc gatctggtcc ttgctattgc
     1981 acccgttctc cgattacgag tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc
     2041 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2101 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2161 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2221 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2281 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2341 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2401 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2461 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     2521 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     2581 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     2641 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     2701 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     2761 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     2821 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     2881 cgttcatcca tagttgcatt taaatttccg aactctccaa ggccctcgtc ggaaaatctt
     2941 caaacctttc gtccgatcca tcttgcaggc tacctctcga acgaactatc gcaagtctct
     3001 tggccggcct tgcgccttgg ctattgcttg gcagcgccta tcgccaggta ttactccaat
     3061 cccgaatatc cgagatcggg atcacccgag agaagttcaa cctacatcct caatcccgat
     3121 ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct ggcctccgcg ccgggttttg
     3181 gcgcctcccg cgggcgcccc cctcgtcacg gcgagcgctg ccacgtcaga cgaagggcgc
     3241 aggagcgtcc tgatccttcc gcccggacgc tcaggacagc ggcccgctgc tcataagact
     3301 cggccttaga accccagtat cagcagaagg acattttagg acgggacttg ggtgactcta
     3361 gggcactggt tttctttcca gagagcggaa caggcgagga aaagtagtcc cttctcggcg
     3421 attctgcgga gggatctccg tggggcggtg aacgccgatg attatataag gacgcgccgg
     3481 gtgtggcaca gctagttccg tcgcagccgg gatttgggtc gcggttcttg tttgtggatc
     3541 gctgtgatcg tcacttggtg agtagcgggc tgctgggctg gccggggctt tcgtggccgc
     3601 cgggccgctc ggtgggacgg aagcgtgtgg agagaccgcc aagggctgta gtctgggtcc
     3661 gcgagcaagg ttgccctgaa ctgggggttg gggggagcgc agcaaaatgg cggctgttcc
     3721 cgagtcttga atggaagacg cttgtgaggc gggctgtgag gtcgttgaaa caaggtgggg
     3781 ggcatggtgg gcggcaagaa cccaaggtct tgagcccttc gctaatgcgg gaaagctctt
     3841 attcgggtga gatgggctgg gcaccatctg gggaccctga cgtgaagttt gtcactgact
     3901 ggagaactcg gtttgtcgtc tgttgcgggg gcggcagtta tggcggtgcc gttgggcagt
     3961 gcacccgtac ctttgggagc gcgcgccctc gtcgtgtcgt gacgtcaccc gttctgttgg
     4021 cttataatgc agggtggggc cacctgccgg taggtgtgcg gtaggctttt ctccgtcgca
     4081 ggacgcaggg ttcgggccta gggtaggctc tcctgaatcg acaggcgccg gacctctggt
     4141 gaggggaggg ataagtgagg cgtcagtttc tttggtcggt tttatgtacc tatcttctta
     4201 agtagctgaa gctccggttt tgaactatgc gctcggggtt ggcgagtgtg ttttgtgaag
     4261 ttttttaggc accttttgaa atgtaatcat ttgggtcaat atgtaatttt cagtgttaga
     4321 cttgtaaatt gtccgctaaa ttctggccgt ttttggcttt tttgttagac aacatgaccg
     4381 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
     4441 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
     4501 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     4561 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     4621 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     4681 tggccgcgca gcaacagatg gagggcctcc tggcgccgca ccggcccaag gagcccgcgt
     4741 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     4801 tcgtgctccc cggagtggag gccgccgagc gcgccggggt gcccgccttc ctggagacct
     4861 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     4921 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc taggctagta
     4981 tgtaagccta gtcttagata ataaaatcgc tatccatcga agatggatgt gtgttggttt
     5041 tttgtgtgtg taacgctagg cgcgcctggt gtaccgagaa cgatcctctc agtgcgagtc
     5101 tcgacgatcc atatcgttgc ttggcagtca gccagtcgga atccagcttg ggacccagga
     5161 agtccaatcg tcagatattg tactcaagcc tggtcacggc agcgtaccga tctgtttaaa
     5221 cctagatatt gatagtctga tcggtcaacg tataatcgag tcctagcttt tgcaaacatc
     5281 tatcaagaga caggatcagc aggaggcttt cgcatgagta ttcaacattt ccgtgtcgcc
     5341 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5401 aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg gttacatcga actggatctc
     5461 aacagcggta agatccttga gagttttcgc cccgaagaac gctttccaat gatgagcact
     5521 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     5581 ggtcgccgca tacactattc tcagaatgac ttggttgagt attcaccagt cacagaaaag
     5641 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5701 aacactgcgg ccaacttact tctgacaacg attggaggac cgaaggagct aaccgctttt
     5761 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5821 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aaccttgcgt
     5881 aaactattaa ctggcgaact acttactcta gcttcccggc aacagttgat agactggatg
     5941 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     6001 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     6061 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     6121 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaaccgatt
     6181 ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac
     6241 atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct
     6301 ccttaccaga aatttatcct taagatcccg aatcgtttaa actcgactct ggctctatcg
     6361 aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc
//
