LOCUS       sequence-307363-        6410 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 6410)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6410)
  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-m4252867
FEATURES             Location/Qualifiers
     source          1..6410
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        58..361
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        363..565
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     516..536
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             865..885
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             889..954
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1007..1128
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1018..1037)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1072..1091
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1243..1289
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1319..1455
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1500..1571
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1797..2385)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1877..1896)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2885..4094
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3931..3950
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4098..4697
                     /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(4098..4117)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4594..4614
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4724..4772
                     /note="synthetic polyadenylation signal"
     CDS             join(5038..5106,5107..5898)
                     /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(5256..5275)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5911..6047
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6199..6270
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6275..6321)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6339..6387
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 ccagatctag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
       61 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      121 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      181 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      241 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      301 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      361 gctgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
      421 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      481 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
      541 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatcagat ctttgtcgat
      601 cctaccatcc actcgacaca cccgccagcg gccgccacca tgtacagctt tgtatctgaa
      661 gagacaggca ctctcatcgt taattcagtg ttgctgttcc tggcatttgt ggtcttcctt
      721 ttagtgaccc tagcgatact gacggccctc cggctgtgcg cttattgttg taacattgtc
      781 aacgttagtc tcgtgaagcc ctcgttctac gtttattccc gcgtgaaaaa tctaaacagc
      841 tcccgtgtgc ctgatcttct tgtcgaaaac ctttactttc agtcgatgga ttataaggac
      901 cacgatggag actacaaaga tcatgacatt gattacaagg acgacgatga caaataagct
      961 agcttgactg actgagatac agcgtacctt cagctcacag acatgataag atacattgat
     1021 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     1081 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     1141 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     1201 aacctctaca aatgtggtat tggcccatct ctatcggtat cgtagcataa ccccttgggg
     1261 cctctaaacg ggtcttgagg ggttttttgt gcccctcggg ccggattgct atctaccggc
     1321 attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca
     1381 gattcagcaa cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca
     1441 gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat ctctcgattt cgatcaagac
     1501 attcctttaa tggtcttttc tggacaccac taggggtcag aagtagttca tcaaactttc
     1561 ttccctccct aatctcattg gttaccttgg gctatcgaaa cttaattaac cagtcaagtc
     1621 agctacttgg cgagatcgac ttgtctgggt ttcgactacg ctcagaattg cgtcagtcaa
     1681 gttcgatctg gtccttgcta ttgcacccgt tctccgatta cgagtttcat ttaaatcatg
     1741 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     1801 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     1861 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     1921 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     1981 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2041 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2101 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     2161 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     2221 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     2281 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     2341 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     2401 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     2461 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     2521 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     2581 cctatctcag cgatctgtct atttcgttca tccatagttg catttaaatt tccgaactct
     2641 ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga tccatcttgc aggctacctc
     2701 tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc ttggctattg cttggcagcg
     2761 cctatcgcca ggtattactc caatcccgaa tatccgagat cgggatcacc cgagagaagt
     2821 tcaacctaca tcctcaatcc cgatctatcc gagatccgag gaatatcgaa atcggggcgc
     2881 gcctggcctc cgcgccgggt tttggcgcct cccgcgggcg cccccctcgt cacggcgagc
     2941 gctgccacgt cagacgaagg gcgcaggagc gtcctgatcc ttccgcccgg acgctcagga
     3001 cagcggcccg ctgctcataa gactcggcct tagaacccca gtatcagcag aaggacattt
     3061 taggacggga cttgggtgac tctagggcac tggttttctt tccagagagc ggaacaggcg
     3121 aggaaaagta gtcccttctc ggcgattctg cggagggatc tccgtggggc ggtgaacgcc
     3181 gatgattata taaggacgcg ccgggtgtgg cacagctagt tccgtcgcag ccgggatttg
     3241 ggtcgcggtt cttgtttgtg gatcgctgtg atcgtcactt ggtgagtagc gggctgctgg
     3301 gctggccggg gctttcgtgg ccgccgggcc gctcggtggg acggaagcgt gtggagagac
     3361 cgccaagggc tgtagtctgg gtccgcgagc aaggttgccc tgaactgggg gttgggggga
     3421 gcgcagcaaa atggcggctg ttcccgagtc ttgaatggaa gacgcttgtg aggcgggctg
     3481 tgaggtcgtt gaaacaaggt ggggggcatg gtgggcggca agaacccaag gtcttgagcc
     3541 cttcgctaat gcgggaaagc tcttattcgg gtgagatggg ctgggcacca tctggggacc
     3601 ctgacgtgaa gtttgtcact gactggagaa ctcggtttgt cgtctgttgc gggggcggca
     3661 gttatggcgg tgccgttggg cagtgcaccc gtacctttgg gagcgcgcgc cctcgtcgtg
     3721 tcgtgacgtc acccgttctg ttggcttata atgcagggtg gggccaccag ccggtaggtg
     3781 tgcggtaggc ttttctccgt cgcaggacgc agggttcggg cctagggtag gctctcctga
     3841 atcgacaggc gccggacctc tggtgagggg agggataagt gaggcgtcag tttctttggt
     3901 cggttttatg tacctatctt cttaagtagc tgaagctccg gttttgaact atgcgctcgg
     3961 ggttggcgag tgtgttttgt gaagtttttt aggcaccttt tgaaatgtaa tcatttgggt
     4021 caatatgtaa ttttcagtgt tagacttgta aattgtccgc taaattctgg ccgtttttgg
     4081 cttttttgtt agacaacatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
     4141 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
     4201 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
     4261 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg
     4321 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
     4381 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggagggc ctcctggcgc
     4441 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc
     4501 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggccgcc gagcgcgccg
     4561 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
     4621 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
     4681 gcaagcccgg tgcctaggct agtatgtaag cctagtctta gataataaaa tcgctatcca
     4741 tcgaagatgg atgtgtgttg gttttttgtg tgtgtaacgc taggcgcgcc tggtgtaccg
     4801 agaacgatcc tctcagtgcg agtctcgacg atccatatcg ttgcttggca gtcagccagt
     4861 cggaatccag cttgggaccc aggaagtcca atcgtcagat attgtactca agcctggtca
     4921 cggcagcgta ccgatctgtt taaacctaga tattgatagt ctgatcggtc aacgtataat
     4981 cgagtcctag cttttgcaaa catctatcaa gagacaggat cagcaggagg ctttcgcatg
     5041 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5101 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcgcga
     5161 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5221 gaacgctttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5281 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5341 gagtattcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5401 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgattgga
     5461 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5521 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5581 gtagcaatgg caacaacctt gcgtaaacta ttaactggcg aactacttac tctagcttcc
     5641 cggcaacagt tgatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5701 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5761 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5821 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5881 ctgattaagc attggtaacc gattctaggt gcattggcgc agaaaaaaat gcctgatgcg
     5941 acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa
     6001 cttgcccact tccatacgtg tcctccttac cagaaattta tccttaagat cccgaatcgt
     6061 ttaaactcga ctctggctct atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg
     6121 gcgctcattt gctcgtcggg catcgaatct cgtcagctat cgtcagctta cctttttggc
     6181 agcgatcgcg gctcccgaca tcttggacca ttagctccac aggtatcttc ttccctctag
     6241 tggtcataac agcagcttca gctacctctc aattcaaaaa acccctcaag acccgtttag
     6301 aggccccaag gggttatgct atcaatcgtt gcgttacaca cacaaaaaac caacacacat
     6361 ccatcttcga tggatagcga ttttattatc taactgctga tcgagtgtag
//
