LOCUS       sequence-297924-        4503 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 4503)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4503)
  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-m4134229
FEATURES             Location/Qualifiers
     source          1..4503
                     /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             254..319
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    372..493
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(383..402)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     437..456
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      608..654
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      684..820
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    865..936
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1162..1750)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1242..1261)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2496..2564,2565..3356)
                     /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(2714..2733)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3369..3505
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3657..3728
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3733..3779)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3797..3845
                     /note="synthetic polyadenylation signal"
     enhancer        3926..4229
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4231..4433
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4384..4404
                     /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 atggattata aggaccacga tggagactac aaagatcatg acattgatta
      301 caaggacgac gatgacaaat aagctagctt gactgactga gatacagcgt accttcagct
      361 cacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
      421 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
      481 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
      541 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtattggcc catctctatc
      601 ggtatcgtag cataacccct tggggcctct aaacgggtct tgaggggttt tttgtgcccc
      661 tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc
      721 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc
      781 acttccatac gtgtcctcct taccagaaat ttatccttaa ggtcgtcagc tatcctgcag
      841 gcgatctctc gatttcgatc aagacattcc tttaatggtc ttttctggac accactaggg
      901 gtcagaagta gttcatcaaa ctttcttccc tccctaatct cattggttac cttgggctat
      961 cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga tcgacttgtc tgggtttcga
     1021 ctacgctcag aattgcgtca gtcaagttcg atctggtcct tgctattgca cccgttctcc
     1081 gattacgagt ttcatttaaa tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     1141 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     1201 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     1261 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     1321 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     1381 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     1441 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     1501 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     1561 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     1621 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     1681 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     1741 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     1801 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     1861 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     1921 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     1981 agttgcattt aaatttccga actctccaag gccctcgtcg gaaaatcttc aaacctttcg
     2041 tccgatccat cttgcaggct acctctcgaa cgaactatcg caagtctctt ggccggcctt
     2101 gcgccttggc tattgcttgg cagcgcctat cgccaggtat tactccaatc ccgaatatcc
     2161 gagatcggga tcacccgaga gaagttcaac ctacatcctc aatcccgatc tatccgagat
     2221 ccgaggaata tcgaaatcgg ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag
     2281 tctcgacgat ccatatcgtt gcttggcagt cagccagtcg gaatccagct tgggacccag
     2341 gaagtccaat cgtcagatat tgtactcaag cctggtcacg gcagcgtacc gatctgttta
     2401 aacctagata ttgatagtct gatcggtcaa cgtataatcg agtcctagct tttgcaaaca
     2461 tctatcaaga gacaggatca gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg
     2521 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     2581 tgaaagtaaa agatgctgaa gatcagttgg gtgcgcgagt gggttacatc gaactggatc
     2641 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgctttcca atgatgagca
     2701 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     2761 tcggtcgccg catacactat tctcagaatg acttggttga gtattcacca gtcacagaaa
     2821 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     2881 ataacactgc ggccaactta cttctgacaa cgattggagg accgaaggag ctaaccgctt
     2941 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3001 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaaccttgc
     3061 gtaaactatt aactggcgaa ctacttactc tagcttcccg gcaacagttg atagactgga
     3121 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     3181 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     3241 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     3301 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaaccga
     3361 ttctaggtgc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc
     3421 acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc
     3481 ctccttacca gaaatttatc cttaagatcc cgaatcgttt aaactcgact ctggctctat
     3541 cgaatctccg tcgtttcgag cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca
     3601 tcgaatctcg tcagctatcg tcagcttacc tttttggcag cgatcgcggc tcccgacatc
     3661 ttggaccatt agctccacag gtatcttctt ccctctagtg gtcataacag cagcttcagc
     3721 tacctctcaa ttcaaaaaac ccctcaagac ccgtttagag gccccaaggg gttatgctat
     3781 caatcgttgc gttacacaca caaaaaacca acacacatcc atcttcgatg gatagcgatt
     3841 ttattatcta actgctgatc gagtgtagcc agatctagta atcaattacg gggtcattag
     3901 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     3961 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     4021 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     4081 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     4141 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     4201 tctacgtatt agtcatcgct attaccatgc tgatgcggtt ttggcagtac atcaatgggc
     4261 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     4321 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     4381 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
     4441 tgaaccgtca gatcagatct ttgtcgatcc taccatccac tcgacacacc cgccagcggc
     4501 cgc
//
