LOCUS       sequence-307314-        6731 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 6731)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6731)
  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-m4252807
FEATURES             Location/Qualifiers
     source          1..6731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        499..1708
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     1545..1564
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             1712..2311
                     /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(1712..1731)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2208..2228
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    2338..2386
                     /note="synthetic polyadenylation signal"
     CDS             join(2652..2720,2721..3512)
                     /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(2870..2889)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3525..3661
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3813..3884
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3889..3935)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3953..4001
                     /note="synthetic polyadenylation signal"
     enhancer        4082..4385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4387..4589
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4540..4560
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             4889..4909
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             5258..5275
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5352..5473
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5363..5382)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5417..5436
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      5588..5634
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      5664..5800
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5845..5916
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(6142..6730)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6222..6241)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
       61 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
      121 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
      181 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcattta
      241 aatttccgaa ctctccaagg ccctcgtcgg aaaatcttca aacctttcgt ccgatccatc
      301 ttgcaggcta cctctcgaac gaactatcgc aagtctcttg gccggccttg cgccttggct
      361 attgcttggc agcgcctatc gccaggtatt actccaatcc cgaatatccg agatcgggat
      421 cacccgagag aagttcaacc tacatcctca atcccgatct atccgagatc cgaggaatat
      481 cgaaatcggg gcgcgcctgg cctccgcgcc gggttttggc gcctcccgcg ggcgcccccc
      541 tcgtcacggc gagcgctgcc acgtcagacg aagggcgcag gagcgtcctg atccttccgc
      601 ccggacgctc aggacagcgg cccgctgctc ataagactcg gccttagaac cccagtatca
      661 gcagaaggac attttaggac gggacttggg tgactctagg gcactggttt tctttccaga
      721 gagcggaaca ggcgaggaaa agtagtccct tctcggcgat tctgcggagg gatctccgtg
      781 gggcggtgaa cgccgatgat tatataagga cgcgccgggt gtggcacagc tagttccgtc
      841 gcagccggga tttgggtcgc ggttcttgtt tgtggatcgc tgtgatcgtc acttggtgag
      901 tagcgggctg ctgggctggc cggggctttc gtggccgccg ggccgctcgg tgggacggaa
      961 gcgtgtggag agaccgccaa gggctgtagt ctgggtccgc gagcaaggtt gccctgaact
     1021 gggggttggg gggagcgcag caaaatggcg gctgttcccg agtcttgaat ggaagacgct
     1081 tgtgaggcgg gctgtgaggt cgttgaaaca aggtgggggg catggtgggc ggcaagaacc
     1141 caaggtcttg agcccttcgc taatgcggga aagctcttat tcgggtgaga tgggctgggc
     1201 accatctggg gaccctgacg tgaagtttgt cactgactgg agaactcggt ttgtcgtctg
     1261 ttgcgggggc ggcagttatg gcggtgccgt tgggcagtgc acccgtacct ttgggagcgc
     1321 gcgccctcgt cgtgtcgtga cgtcacccgt tctgttggct tataatgcag ggtggggcca
     1381 ccagccggta ggtgtgcggt aggcttttct ccgtcgcagg acgcagggtt cgggcctagg
     1441 gtaggctctc ctgaatcgac aggcgccgga cctctggtga ggggagggat aagtgaggcg
     1501 tcagtttctt tggtcggttt tatgtaccta tcttcttaag tagctgaagc tccggttttg
     1561 aactatgcgc tcggggttgg cgagtgtgtt ttgtgaagtt ttttaggcac cttttgaaat
     1621 gtaatcattt gggtcaatat gtaattttca gtgttagact tgtaaattgt ccgctaaatt
     1681 ctggccgttt ttggcttttt tgttagacaa catgaccgag tacaagccca cggtgcgcct
     1741 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta
     1801 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca
     1861 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
     1921 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
     1981 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
     2041 gggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
     2101 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
     2161 cgccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
     2221 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
     2281 ctggtgcatg acccgcaagc ccggtgccta ggctagtatg taagcctagt cttagataat
     2341 aaaatcgcta tccatcgaag atggatgtgt gttggttttt tgtgtgtgta acgctaggcg
     2401 cgcctggtgt accgagaacg atcctctcag tgcgagtctc gacgatccat atcgttgctt
     2461 ggcagtcagc cagtcggaat ccagcttggg acccaggaag tccaatcgtc agatattgta
     2521 ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc tagatattga tagtctgatc
     2581 ggtcaacgta taatcgagtc ctagcttttg caaacatcta tcaagagaca ggatcagcag
     2641 gaggctttcg catgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     2701 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     2761 agttgggtgc gcgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     2821 gttttcgccc cgaagaacgc tttccaatga tgagcacttt taaagttctg ctatgtggcg
     2881 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     2941 agaatgactt ggttgagtat tcaccagtca cagaaaagca tcttacggat ggcatgacag
     3001 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3061 tgacaacgat tggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3121 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     3181 acaccacgat gcctgtagca atggcaacaa ccttgcgtaa actattaact ggcgaactac
     3241 ttactctagc ttcccggcaa cagttgatag actggatgga ggcggataaa gttgcaggac
     3301 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     3361 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     3421 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     3481 agataggtgc ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa
     3541 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     3601 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     3661 agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta
     3721 cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag
     3781 cttacctttt tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat
     3841 cttcttccct ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct
     3901 caagacccgt ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa
     3961 aaaccaacac acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt
     4021 gtagccagat ctagtaatca attacggggt cattagttca tagcccatat atggagttcc
     4081 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     4141 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     4201 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     4261 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     4321 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     4381 ccatgctgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     4441 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     4501 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     4561 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc agatctttgt
     4621 cgatcctacc atccactcga cacacccgcc agcggccgcc accatgtact cattcgtttc
     4681 ggaagagaca ggtacgttaa tagttaatag cgtacttctt tttcttgctt tcgtggtatt
     4741 cttgctagtt acactagcca tccttactgc gcttcgattg tgtgcgtact gctgcaatat
     4801 tgttaacgtg agtcttgtaa aaccttcttt ttacgtttac tctcgtgtta aaaatctgaa
     4861 ttcttctaga gttcctgatc ttctggtcga aaacctttac tttcagtcga tgagtgacaa
     4921 aatcattcat ctaaccgatg actcctttga tacagacgtg ttgaaggccg atggtgcgat
     4981 actggtagat ttctgggctg aatggtgtgg cccgtgcaag atgatcgcac ccattctgga
     5041 cgagatcgcc gacgagtatc aaggaaaatt aactgtcgct aagcttaata tagatcagaa
     5101 cccagggacg gcacctaaat acggcatcag gggaatacct accctcctgc tctttaagaa
     5161 tggggaagtt gccgcgacaa aagtgggagc cctgtctaag ggccagctaa aagagttctt
     5221 ggacgctaac ctggcaggtt caggctcggg gcacatgcac catcaccatc accatagcag
     5281 cgggctcgtg ccacgcggct aagctagctt gactgactga gatacagcgt accttcagct
     5341 cacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     5401 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     5461 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     5521 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtattggcc catctctatc
     5581 ggtatcgtag cataacccct tggggcctct aaacgggtct tgaggggttt tttgtgcccc
     5641 tcgggccgga ttgctatcta ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc
     5701 gcgtcttata ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc
     5761 acttccatac gtgtcctcct taccagaaat ttatccttaa ggtcgtcagc tatcctgcag
     5821 gcgatctctc gatttcgatc aagacattcc tttaatggtc ttttctggac accactaggg
     5881 gtcagaagta gttcatcaaa ctttcttccc tccctaatct cattggttac cttgggctat
     5941 cgaaacttaa ttaaccagtc aagtcagcta cttggcgaga tcgacttgtc tgggtttcga
     6001 ctacgctcag aattgcgtca gtcaagttcg atctggtcct tgctattgca cccgttctcc
     6061 gattacgagt ttcatttaaa tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6121 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6181 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6241 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6301 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6361 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6421 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6481 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6541 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     6601 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6661 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6721 aggatctcaa g
//
