LOCUS       sequence-307324-        4452 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 4452)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4452)
  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-m4133668
FEATURES             Location/Qualifiers
     source          1..4452
                     /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"
     CDS             join(2445..2513,2514..3305)
                     /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(2663..2682)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3318..3454
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3606..3677
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3682..3728)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3746..3794
                     /note="synthetic polyadenylation signal"
     enhancer        3875..4178
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4180..4382
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4333..4353
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgtac agctttgtat ctgaagagac aggcactctc atcgttaatt cagtgttgct
       61 gttcctggca tttgtggtct tccttttagt gaccctagcg atactgacgg ccctccggct
      121 gtgcgcttat tgttgtaaca ttgtcaacgt tagtctcgtg aagccctcgt tctacgttta
      181 ttcccgcgtg aaaaatctaa acagctcccg tgtgcctgat cttcttgtcg 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 gtcgtcagct
      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 tgtaccgaga acgatcctct
     2221 cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt
     2281 gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg
     2341 atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt
     2401 ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt
     2461 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     2521 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg
     2581 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa
     2641 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     2701 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag
     2761 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     2821 ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc
     2881 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     2941 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     3001 caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg caacagttga
     3061 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     3121 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3181 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     3241 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     3301 ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
     3361 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
     3421 atacgtgtcc tccttaccag aaatttatcc ttaagatccc gaatcgttta aactcgactc
     3481 tggctctatc gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg ctcatttgct
     3541 cgtcgggcat cgaatctcgt cagctatcgt cagcttacct ttttggcagc gatcgcggct
     3601 cccgacatct tggaccatta gctccacagg tatcttcttc cctctagtgg tcataacagc
     3661 agcttcagct acctctcaat tcaaaaaacc cctcaagacc cgtttagagg ccccaagggg
     3721 ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa cacacatcca tcttcgatgg
     3781 atagcgattt tattatctaa ctgctgatcg agtgtagcca gatctagtaa tcaattacgg
     3841 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     3901 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     3961 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     4021 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     4081 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     4141 ggcagtacat ctacgtatta gtcatcgcta ttaccatgct gatgcggttt tggcagtaca
     4201 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     4261 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     4321 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     4381 ctggtttagt gaaccgtcag atcagatctt tgtcgatcct accatccact cgacacaccc
     4441 gccagcggcc gc
//
