LOCUS       sequence-300639-        6359 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 6359)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6359)
  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-m4252835
FEATURES             Location/Qualifiers
     source          1..6359
                     /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..268
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    321..442
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(332..351)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     386..405
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      557..603
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      633..769
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    814..885
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1111..1699)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1191..1210)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2199..3408
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3245..3264
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3412..4011
                     /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(3412..3431)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3908..3928
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4038..4086
                     /note="synthetic polyadenylation signal"
     CDS             join(4352..4420,4421..5212)
                     /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(4570..4589)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5225..5361
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5513..5584
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5589..5635)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5653..5701
                     /note="synthetic polyadenylation signal"
     enhancer        5782..6085
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6087..6289
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6240..6260
                     /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 caccatcacc atcaccatta agctagcttg actgactgag atacagcgta
      301 ccttcagctc acagacatga taagatacat tgatgagttt ggacaaacca caactagaat
      361 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
      421 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
      481 gggggaggtg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc
      541 atctctatcg gtatcgtagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
      601 ttgtgcccct cgggccggat tgctatctac cggcattggc gcagaaaaaa atgcctgatg
      661 cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata cggcttcccc
      721 aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag gtnnnnnnnn
      781 atcctgcagg cgatctctcg atttcgatca agacattcct ttaatggtct tttctggaca
      841 ccactagggg tcagaagtag ttcatcaaac tttcttccct ccctaatctc attggttacc
      901 ttgggctatc gaaacttaat taaccagtca agtcagctac ttggcgagat cgacttgtct
      961 gggtttcgac tacgctcaga attgcgtcag tcaagttcga tctggtcctt gctattgcac
     1021 ccgttctccg attacgagtt tcatttaaat catgtgagca aaaggccagc aaaaggccag
     1081 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     1141 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     1201 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     1261 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     1321 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     1381 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     1441 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     1501 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     1561 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     1621 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     1681 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     1741 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     1801 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     1861 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     1921 ttcatccata gttgcattta aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca
     1981 aacctttcgt ccgatccatc ttgcaggcta cctctcgaac gaactatcgc aagtctcttg
     2041 gccggccttg cgccttggct attgcttggc agcgcctatc gccaggtatt actccaatcc
     2101 cgaatatccg agatcgggat cacccgagag aagttcaacc tacatcctca atcccgatct
     2161 atccgagatc cgaggaatat cgaaatcggg gcgcgcctgg cctccgcgcc gggttttggc
     2221 gcctcccgcg ggcgcccccc tcgtcacggc gagcgctgcc acgtcagacg aagggcgcag
     2281 gagcgtcctg atccttccgc ccggacgctc aggacagcgg cccgctgctc ataagactcg
     2341 gccttagaac cccagtatca gcagaaggac attttaggac gggacttggg tgactctagg
     2401 gcactggttt tctttccaga gagcggaaca ggcgaggaaa agtagtccct tctcggcgat
     2461 tctgcggagg gatctccgtg gggcggtgaa cgccgatgat tatataagga cgcgccgggt
     2521 gtggcacagc tagttccgtc gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc
     2581 tgtgatcgtc acttggtgag tagcgggctg ctgggctggc cggggctttc gtggccgccg
     2641 ggccgctcgg tgggacggaa gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc
     2701 gagcaaggtt gccctgaact gggggttggg gggagcgcag caaaatggcg gctgttcccg
     2761 agtcttgaat ggaagacgct tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg
     2821 catggtgggc ggcaagaacc caaggtcttg agcccttcgc taatgcggga aagctcttat
     2881 tcgggtgaga tgggctgggc accatctggg gaccctgacg tgaagtttgt cactgactgg
     2941 agaactcggt ttgtcgtctg ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc
     3001 acccgtacct ttgggagcgc gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct
     3061 tataatgcag ggtggggcca ccagccggta ggtgtgcggt aggcttttct ccgtcgcagg
     3121 acgcagggtt cgggcctagg gtaggctctc ctgaatcgac aggcgccgga cctctggtga
     3181 ggggagggat aagtgaggcg tcagtttctt tggtcggttt tatgtaccta tcttcttaag
     3241 tagctgaagc tccggttttg aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt
     3301 ttttaggcac cttttgaaat gtaatcattt gggtcaatat gtaattttca gtgttagact
     3361 tgtaaattgt ccgctaaatt ctggccgttt ttggcttttt tgttagacaa catgaccgag
     3421 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc
     3481 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc
     3541 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     3601 gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa
     3661 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     3721 gccgcgcagc aacagatgga gggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     3781 ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     3841 gtgctccccg gagtggaggc cgccgagcgc gccggggtgc ccgccttcct ggagacctcc
     3901 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     3961 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgccta ggctagtatg
     4021 taagcctagt cttagataat aaaatcgcta tccatcgaag atggatgtgt gttggttttt
     4081 tgtgtgtgta acgctaggcg cgcctggtgt accgagaacg atcctctcag tgcgagtctc
     4141 gacgatccat atcgttgctt ggcagtcagc cagtcggaat ccagcttggg acccaggaag
     4201 tccaatcgtc agatattgta ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc
     4261 tagatattga tagtctgatc ggtcaacgta taatcgagtc ctagcttttg caaacatcta
     4321 tcaagagaca ggatcagcag gaggctttcg catgagtatt caacatttcc gtgtcgccct
     4381 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     4441 agtaaaagat gctgaagatc agttgggtgc gcgagtgggt tacatcgaac tggatctcaa
     4501 cagcggtaag atccttgaga gttttcgccc cgaagaacgc tttccaatga tgagcacttt
     4561 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     4621 tcgccgcata cactattctc agaatgactt ggttgagtat tcaccagtca cagaaaagca
     4681 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     4741 cactgcggcc aacttacttc tgacaacgat tggaggaccg aaggagctaa ccgctttttt
     4801 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     4861 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa ccttgcgtaa
     4921 actattaact ggcgaactac ttactctagc ttcccggcaa cagttgatag actggatgga
     4981 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     5041 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     5101 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     5161 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aaccgattct
     5221 aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat
     5281 atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc
     5341 ttaccagaaa tttatcctta agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa
     5401 tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga
     5461 atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc gacatcttgg
     5521 accattagct ccacaggtat cttcttccct ctagtggtca taacagcagc ttcagctacc
     5581 tctcaattca aaaaacccct caagacccgt ttagaggccc caaggggtta tgctatcaat
     5641 cgttgcgtta cacacacaaa aaaccaacac acatccatct tcgatggata gcgattttat
     5701 tatctaactg ctgatcgagt gtagccagat ctagtaatca attacggggt cattagttca
     5761 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     5821 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     5881 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     5941 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     6001 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     6061 cgtattagtc atcgctatta ccatgctgat gcggttttgg cagtacatca atgggcgtgg
     6121 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     6181 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     6241 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
     6301 ccgtcagatc agatctttgt cgatcctacc atccactcga cacacccgcc agcggccgc
//
