LOCUS       sequence-307364-        5316 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 E (envelope).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5316)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5316)
  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-m4252822
FEATURES             Location/Qualifiers
     source          1..5316
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     polyA_signal    11..132
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(22..41)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     76..95
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      247..293
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      323..459
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    504..575
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(801..1389)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(881..900)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1889..3098
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     2935..2954
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3102..3701
                     /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(3102..3121)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3598..3618
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3728..3776
                     /note="synthetic polyadenylation signal"
     misc_feature    4162..4233
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4238..4284)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4302..4350
                     /note="synthetic polyadenylation signal"
     enhancer        4431..4734
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4735..4938
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4889..4909
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             5238..5258
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             5259..5303
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
ORIGIN
        1 acagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
       61 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
      121 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
      181 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc atctctatcg
      241 gtatcgtagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgtgcccct
      301 cgggccggat tgctatctac cggcattggc gcagaaaaaa atgcctgatg cgacgctgcg
      361 cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca
      421 cttccatacg tgtcctcctt accagaaatt tatccttaag gtcgtcagct atcctgcagg
      481 cgatctctcg atttcgatca agacattcct ttaatggtct tttctggaca ccactagggg
      541 tcagaagtag ttcatcaaac tttcttccct ccctaatctc attggttacc ttgggctatc
      601 gaaacttaat taaccagtca agtcagctac ttggcgagat cgacttgtct gggtttcgac
      661 tacgctcaga attgcgtcag tcaagttcga tctggtcctt gctattgcac ccgttctccg
      721 attacgagtt tcatttaaat catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
      781 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
      841 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
      901 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
      961 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     1021 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     1081 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     1141 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     1201 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     1261 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     1321 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     1381 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     1441 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     1501 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     1561 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     1621 gttgcattta aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt
     1681 ccgatccatc ttgcaggcta cctctcgaac gaactatcgc aagtctcttg gccggccttg
     1741 cgccttggct attgcttggc agcgcctatc gccaggtatt actccaatcc cgaatatccg
     1801 agatcgggat cacccgagag aagttcaacc tacatcctca atcccgatct atccgagatc
     1861 cgaggaatat cgaaatcggg gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg
     1921 ggcgcccccc tcgtcacggc gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg
     1981 atccttccgc ccggacgctc aggacagcgg cccgctgctc ataagactcg gccttagaac
     2041 cccagtatca gcagaaggac attttaggac gggacttggg tgactctagg gcactggttt
     2101 tctttccaga gagcggaaca ggcgaggaaa agtagtccct tctcggcgat tctgcggagg
     2161 gatctccgtg gggcggtgaa cgccgatgat tatataagga cgcgccgggt gtggcacagc
     2221 tagttccgtc gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc
     2281 acttggtgag tagcgggctg ctgggctggc cggggctttc gtggccgccg ggccgctcgg
     2341 tgggacggaa gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt
     2401 gccctgaact gggggttggg gggagcgcag caaaatggcg gctgttcccg agtcttgaat
     2461 ggaagacgct tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg catggtgggc
     2521 ggcaagaacc caaggtcttg agcccttcgc taatgcggga aagctcttat tcgggtgaga
     2581 tgggctgggc accatctggg gaccctgacg tgaagtttgt cactgactgg agaactcggt
     2641 ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc acccgtacct
     2701 ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct tataatgcag
     2761 ggtggggcca ccagccggta ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt
     2821 cgggcctagg gtaggctctc ctgaatcgac aggcgccgga cctctggtga ggggagggat
     2881 aagtgaggcg tcagtttctt tggtcggttt tatgtaccta tcttcttaag tagctgaagc
     2941 tccggttttg aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac
     3001 cttttgaaat gtaatcattt gggtcaatat gtaattttca gtgttagact tgtaaattgt
     3061 ccgctaaatt ctggccgttt ttggcttttt tgttagacaa catgaccgag tacaagccca
     3121 cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt
     3181 tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca
     3241 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     3301 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     3361 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     3421 aacagatgga gggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     3481 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     3541 gagtggaggc cgccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     3601 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     3661 gaccgcgcac ctggtgcatg acccgcaagc ccggtgccta ggctagtatg taagcctagt
     3721 cttagataat aaaatcgcta tccatcgaag atggatgtgt gttggttttt tgtgtgtgta
     3781 acgctaggcg cgcctggtgt accgagaacg atcctctcag tgcgagtctc gacgatccat
     3841 atcgttgctt ggcagtcagc cagtcggaat ccagcttggg acccaggaag tccaatcgtc
     3901 agatattgta ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc tagatattga
     3961 tagtctgatc ggtcaacgta taatcgagtc ctagcttttg caaacatcta tcaagagaca
     4021 ggtttaaact cgactctggc tctatcgaat ctccgtcgtt tcgagcttac gcgaacagcc
     4081 gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt
     4141 ggcagcgatc gcggctcccg acatcttgga ccattagctc cacaggtatc ttcttccctc
     4201 tagtggtcat aacagcagct tcagctacct ctcaattcaa aaaacccctc aagacccgtt
     4261 tagaggcccc aaggggttat gctatcaatc gttgcgttac acacacaaaa aaccaacaca
     4321 catccatctt cgatggatag cgattttatt atctaactgc tgatcgagtg tagccagatc
     4381 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     4441 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     4501 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     4561 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     4621 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     4681 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     4741 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     4801 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     4861 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     4921 gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc gatcctacca
     4981 tccactcgac acacccgcca gcggccgcca ccatgtactc attcgtttcg gaagagacag
     5041 gtacgttaat agttaatagc gtacttcttt ttcttgcttt cgtggtattc ttgctagtta
     5101 cactagccat ccttactgcg cttcgattgt gtgcgtactg ctgcaatatt gttaacgtga
     5161 gtcttgtaaa accttctttt tacgtttact ctcgtgttaa aaatctgaat tcttctagag
     5221 ttcctgatct tctggtcgaa aacctttact ttcagtcgaa agaaaccgcg gcagctaagt
     5281 tcgagaggca gcacatggac agttaagcta gcttga
//
