LOCUS       sequence-300600-        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-m4134082
FEATURES             Location/Qualifiers
     source          1..4479
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(185..773)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(265..284)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(1519..1587,1588..2379)
                     /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(1737..1756)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      2392..2528
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2680..2751
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(2756..2802)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    2820..2868
                     /note="synthetic polyadenylation signal"
     enhancer        2949..3252
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3254..3456
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3407..3427
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3756..3776
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3777..3821
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    3874..3995
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3885..3904)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3939..3958
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      4110..4156
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      4186..4322
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4367..4438
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
ORIGIN
        1 gtcaagtcag ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg
       61 tcagtcaagt tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt
      121 aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
      181 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
      241 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
      301 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
      361 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
      421 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
      481 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
      541 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
      601 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
      661 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
      721 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
      781 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
      841 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
      901 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
      961 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc
     1021 cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag
     1081 gctacctctc gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct
     1141 tggcagcgcc tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg
     1201 agagaagttc aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat
     1261 cggggcgcgc ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc
     1321 gttgcttggc agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga
     1381 tattgtactc aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag
     1441 tctgatcggt caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga
     1501 tcagcaggag gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt
     1561 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     1621 gaagatcagt tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc
     1681 cttgagagtt ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta
     1741 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     1801 tattctcaga atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc
     1861 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     1921 ttacttctga caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     1981 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     2041 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc
     2101 gaactactta ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt
     2161 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     2221 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     2281 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     2341 atcgctgaga taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg
     2401 cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag
     2461 caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt
     2521 atccttaaga tcccgaatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc
     2581 gagcttacgc gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta
     2641 tcgtcagctt acctttttgg cagcgatcgc ggctcccgac atcttggacc attagctcca
     2701 caggtatctt cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa
     2761 aacccctcaa gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac
     2821 acacaaaaaa ccaacacaca tccatcttcg atggatagcg attttattat ctaactgctg
     2881 atcgagtgta gccagatcta gtaatcaatt acggggtcat tagttcatag cccatatatg
     2941 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     3001 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     3061 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     3121 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     3181 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     3241 gctattacca tgctgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     3301 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     3361 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     3421 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatcaga
     3481 tctttgtcga tcctaccatc cactcgacac acccgccagc ggccgccacc atgtacagct
     3541 ttgtatctga agagacaggc actctcatcg ttaattcagt gttgctgttc ctggcatttg
     3601 tggtcttcct tttagtgacc ctagcgatac tgacggccct ccggctgtgc gcttattgtt
     3661 gtaacattgt caacgttagt ctcgtgaagc cctcgttcta cgtttattcc cgcgtgaaaa
     3721 atctaaacag ctcccgtgtg cctgatcttc ttgtcgaaaa cctttacttt cagtcgaaag
     3781 aaaccgcggc agctaagttc gagaggcagc acatggacag ttaagctagc ttgactgact
     3841 gagatacagc gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa
     3901 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     3961 tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta
     4021 tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat
     4081 gtggtattgg cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt
     4141 cttgaggggt tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa
     4201 aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg
     4261 atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt
     4321 aaggtcgtca gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg
     4381 tcttttctgg acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat
     4441 ctcattggtt accttgggct atcgaaactt aattaacca
//
