LOCUS       sequence-297253-        6386 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 6386)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6386)
  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-m4252821
FEATURES             Location/Qualifiers
     source          1..6386
                     /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..295
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    348..469
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(359..378)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     413..432
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      584..630
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      660..796
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    841..912
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1138..1726)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1218..1237)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2226..3435
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3272..3291
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3439..4038
                     /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(3439..3458)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3935..3955
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4065..4113
                     /note="synthetic polyadenylation signal"
     CDS             join(4379..4447,4448..5239)
                     /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(4597..4616)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5252..5388
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5540..5611
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5616..5662)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5680..5728
                     /note="synthetic polyadenylation signal"
     enhancer        5809..6112
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6114..6316
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6267..6287
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgtac agctttgtat ctgaagagac aggcactctc atcgttaatt cagtgttgct
       61 gttcctggca tttgtggtct tccttttagt gaccctagcg atactgacgg ccctccggct
      121 gtgcgcttat tgttgtaaca ttgtcaacgt tagtctcgtg aagccctcgt tctacgttta
      181 ttcccgcgtg aaaaatctaa acagctcccg tgtgcctgat cttcttgtcg aaaaccttta
      241 ctttcagtcg aaagaaaccg cggcagctaa gttcgagagg cagcacatgg acagttaagc
      301 tagcttgact gactgagata cagcgtacct tcagctcaca gacatgataa gatacattga
      361 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
      421 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
      481 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
      541 aaacctctac aaatgtggta ttggcccatc tctatcggta tcgtagcata accccttggg
      601 gcctctaaac gggtcttgag gggttttttg tgcccctcgg gccggattgc tatctaccgg
      661 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
      721 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
      781 agaaatttat ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga
      841 cattccttta atggtctttt ctggacacca ctaggggtca gaagtagttc atcaaacttt
      901 cttccctccc taatctcatt ggttaccttg ggctatcgaa acttaattaa ccagtcaagt
      961 cagctacttg gcgagatcga cttgtctggg tttcgactac gctcagaatt gcgtcagtca
     1021 agttcgatct ggtccttgct attgcacccg ttctccgatt acgagtttca tttaaatcat
     1081 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1141 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1201 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1261 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1321 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1381 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     1441 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     1501 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     1561 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     1621 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     1681 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     1741 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     1801 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     1861 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     1921 acctatctca gcgatctgtc tatttcgttc atccatagtt gcatttaaat ttccgaactc
     1981 tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg caggctacct
     2041 ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc cttggctatt gcttggcagc
     2101 gcctatcgcc aggtattact ccaatcccga atatccgaga tcgggatcac ccgagagaag
     2161 ttcaacctac atcctcaatc ccgatctatc cgagatccga ggaatatcga aatcggggcg
     2221 cgcctggcct ccgcgccggg ttttggcgcc tcccgcgggc gcccccctcg tcacggcgag
     2281 cgctgccacg tcagacgaag ggcgcaggag cgtcctgatc cttccgcccg gacgctcagg
     2341 acagcggccc gctgctcata agactcggcc ttagaacccc agtatcagca gaaggacatt
     2401 ttaggacggg acttgggtga ctctagggca ctggttttct ttccagagag cggaacaggc
     2461 gaggaaaagt agtcccttct cggcgattct gcggagggat ctccgtgggg cggtgaacgc
     2521 cgatgattat ataaggacgc gccgggtgtg gcacagctag ttccgtcgca gccgggattt
     2581 gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag cgggctgctg
     2641 ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg gacggaagcg tgtggagaga
     2701 ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc ctgaactggg ggttgggggg
     2761 agcgcagcaa aatggcggct gttcccgagt cttgaatgga agacgcttgt gaggcgggct
     2821 gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc aagaacccaa ggtcttgagc
     2881 ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg gctgggcacc atctggggac
     2941 cctgacgtga agtttgtcac tgactggaga actcggtttg tcgtctgttg cgggggcggc
     3001 agttatggcg gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg ccctcgtcgt
     3061 gtcgtgacgt cacccgttct gttggcttat aatgcagggt ggggccacca gccggtaggt
     3121 gtgcggtagg cttttctccg tcgcaggacg cagggttcgg gcctagggta ggctctcctg
     3181 aatcgacagg cgccggacct ctggtgaggg gagggataag tgaggcgtca gtttctttgg
     3241 tcggttttat gtacctatct tcttaagtag ctgaagctcc ggttttgaac tatgcgctcg
     3301 gggttggcga gtgtgttttg tgaagttttt taggcacctt ttgaaatgta atcatttggg
     3361 tcaatatgta attttcagtg ttagacttgt aaattgtccg ctaaattctg gccgtttttg
     3421 gcttttttgt tagacaacat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
     3481 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
     3541 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
     3601 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     3661 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     3721 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaggg cctcctggcg
     3781 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
     3841 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggccgc cgagcgcgcc
     3901 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     3961 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     4021 cgcaagcccg gtgcctaggc tagtatgtaa gcctagtctt agataataaa atcgctatcc
     4081 atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg ctaggcgcgc ctggtgtacc
     4141 gagaacgatc ctctcagtgc gagtctcgac gatccatatc gttgcttggc agtcagccag
     4201 tcggaatcca gcttgggacc caggaagtcc aatcgtcaga tattgtactc aagcctggtc
     4261 acggcagcgt accgatctgt ttaaacctag atattgatag tctgatcggt caacgtataa
     4321 tcgagtccta gcttttgcaa acatctatca agagacagga tcagcaggag gctttcgcat
     4381 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4441 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg
     4501 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     4561 agaacgcttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4621 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4681 tgagtattca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4741 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgattgg
     4801 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4861 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4921 tgtagcaatg gcaacaacct tgcgtaaact attaactggc gaactactta ctctagcttc
     4981 ccggcaacag ttgatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     5041 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     5101 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5161 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     5221 actgattaag cattggtaac cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc
     5281 gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca
     5341 acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaaga tcccgaatcg
     5401 tttaaactcg actctggctc tatcgaatct ccgtcgtttc gagcttacgc gaacagccgt
     5461 ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta tcgtcagctt acctttttgg
     5521 cagcgatcgc ggctcccgac atcttggacc attagctcca caggtatctt cttccctcta
     5581 gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa gacccgttta
     5641 gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa ccaacacaca
     5701 tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta gccagatcta
     5761 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     5821 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     5881 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     5941 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     6001 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     6061 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tgctgatgcg
     6121 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     6181 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     6241 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     6301 ctatataagc agagctggtt tagtgaaccg tcagatcaga tctttgtcga tcctaccatc
     6361 cactcgacac acccgccagc ggccgc
//
