LOCUS       sequence-297381-        4479 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 4479)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4479)
  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-m4133764
FEATURES             Location/Qualifiers
     source          1..4479
                     /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"
     CDS             join(2472..2540,2541..3332)
                     /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(2690..2709)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3345..3481
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3633..3704
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3709..3755)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3773..3821
                     /note="synthetic polyadenylation signal"
     enhancer        3902..4205
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4207..4409
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4360..4380
                     /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 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 cgcctggtgt accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt
     2281 ggcagtcagc cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta
     2341 ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc
     2401 ggtcaacgta taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag
     2461 gaggctttcg catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     2521 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     2581 agttgggtgc gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     2641 gttttcgccc cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg
     2701 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     2761 agaatgactt ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag
     2821 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     2881 tgacaacgat tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     2941 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     3001 acaccacgat gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac
     3061 ttactctagc ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac
     3121 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     3181 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     3241 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     3301 agataggtgc ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa
     3361 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     3421 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     3481 agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta
     3541 cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag
     3601 cttacctttt tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat
     3661 cttcttccct ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct
     3721 caagacccgt ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa
     3781 aaaccaacac acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt
     3841 gtagccagat ctagtaatca attacggggt cattagttca tagcccatat atggagttcc
     3901 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     3961 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     4021 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     4081 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     4141 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     4201 ccatgctgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     4261 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     4321 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     4381 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc agatctttgt
     4441 cgatcctacc atccactcga cacacccgcc agcggccgc
//
