LOCUS       sequence-295057-        6889 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 6889)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6889)
  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-m4133287
FEATURES             Location/Qualifiers
     source          1..6889
                     /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             284..361
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             389..409
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    858..979
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(869..888)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     923..942
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1094..1140
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1170..1306
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1351..1422
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1648..2236)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1728..1747)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2736..3945
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3782..3801
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3949..4548
                     /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(3949..3968)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4445..4465
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4575..4623
                     /note="synthetic polyadenylation signal"
     CDS             join(4889..4957,4958..5749)
                     /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(5107..5126)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5762..5898
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6043..6114
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6119..6165)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6183..6231
                     /note="synthetic polyadenylation signal"
     enhancer        6312..6615
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6617..6819
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6770..6790
                     /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 ggacggcgaa tcccggggct gataaatccc tggaaaagga gatggaagaa
      301 gaactttatt gcagtgtctg cggctaaccg gttcaaaaag atcagttcaa gcggcgccct
      361 agattacgac attcctacga ccgcctccga gaatctctat ttccagggtg aatttggctt
      421 ggcacaacat gacgaggccg tcgataacaa gtttaataaa gaacagcaga acgctttcta
      481 tgagatcctt cacttaccaa atctgaacga agagcaacgc aacgctttca ttcagagcct
      541 gaaagacgat ccctcgcagt ctgcaaatct cctggccgaa gccaagaagc tgaatgacgc
      601 acaggctcca aaagttgata acaagtttaa caaggagcag cagaatgctt tctacgagat
      661 attgcacctt cctaatctga atgaggaaca aagaaatgca tttatccagt ccctaaaaga
      721 cgatccatca cagagtgcca acctgcttgc ggaagccaaa aagctcaatg atgcccaggc
      781 cccgaaagta gacgcgaacc atcaataagc tagcttgact gactgagata cagcgtacct
      841 tcagctcaca gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca
      901 gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat
      961 aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg
     1021 ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta ttggcccatc
     1081 tctatcggta tcgtagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     1141 tgcccctcgg gccggattgc tatctaccgg cattggcgca gaaaaaaatg cctgatgcga
     1201 cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac
     1261 ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaaggtc gtcagctatc
     1321 ctgcaggcga tctctcgatt tcgatcaaga cattccttta atggtctttt ctggacacca
     1381 ctaggggtca gaagtagttc atcaaacttt cttccctccc taatctcatt ggttaccttg
     1441 ggctatcgaa acttaattaa ccagtcaagt cagctacttg gcgagatcga cttgtctggg
     1501 tttcgactac gctcagaatt gcgtcagtca agttcgatct ggtccttgct attgcacccg
     1561 ttctccgatt acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa aggccaggaa
     1621 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     1681 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     1741 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     1801 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     1861 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     1921 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     1981 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2041 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     2101 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2161 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2221 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     2281 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     2341 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     2401 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     2461 atccatagtt gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa atcttcaaac
     2521 ctttcgtccg atccatcttg caggctacct ctcgaacgaa ctatcgcaag tctcttggcc
     2581 ggccttgcgc cttggctatt gcttggcagc gcctatcgcc aggtattact ccaatcccga
     2641 atatccgaga tcgggatcac ccgagagaag ttcaacctac atcctcaatc ccgatctatc
     2701 cgagatccga ggaatatcga aatcggggcg cgcctggcct ccgcgccggg ttttggcgcc
     2761 tcccgcgggc gcccccctcg tcacggcgag cgctgccacg tcagacgaag ggcgcaggag
     2821 cgtcctgatc cttccgcccg gacgctcagg acagcggccc gctgctcata agactcggcc
     2881 ttagaacccc agtatcagca gaaggacatt ttaggacggg acttgggtga ctctagggca
     2941 ctggttttct ttccagagag cggaacaggc gaggaaaagt agtcccttct cggcgattct
     3001 gcggagggat ctccgtgggg cggtgaacgc cgatgattat ataaggacgc gccgggtgtg
     3061 gcacagctag ttccgtcgca gccgggattt gggtcgcggt tcttgtttgt ggatcgctgt
     3121 gatcgtcact tggtgagtag cgggctgctg ggctggccgg ggctttcgtg gccgccgggc
     3181 cgctcggtgg gacggaagcg tgtggagaga ccgccaaggg ctgtagtctg ggtccgcgag
     3241 caaggttgcc ctgaactggg ggttgggggg agcgcagcaa aatggcggct gttcccgagt
     3301 cttgaatgga agacgcttgt gaggcgggct gtgaggtcgt tgaaacaagg tggggggcat
     3361 ggtgggcggc aagaacccaa ggtcttgagc ccttcgctaa tgcgggaaag ctcttattcg
     3421 ggtgagatgg gctgggcacc atctggggac cctgacgtga agtttgtcac tgactggaga
     3481 actcggtttg tcgtctgttg cgggggcggc agttatggcg gtgccgttgg gcagtgcacc
     3541 cgtacctttg ggagcgcgcg ccctcgtcgt gtcgtgacgt cacccgttct gttggcttat
     3601 aatgcagggt ggggccacct gccggtaggt gtgcggtagg cttttctccg tcgcaggacg
     3661 cagggttcgg gcctagggta ggctctcctg aatcgacagg cgccggacct ctggtgaggg
     3721 gagggataag tgaggcgtca gtttctttgg tcggttttat gtacctatct tcttaagtag
     3781 ctgaagctcc ggttttgaac tatgcgctcg gggttggcga gtgtgttttg tgaagttttt
     3841 taggcacctt ttgaaatgta atcatttggg tcaatatgta attttcagtg ttagacttgt
     3901 aaattgtccg ctaaattctg gccgtttttg gcttttttgt tagacaacat gaccgagtac
     3961 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
     4021 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
     4081 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     4141 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
     4201 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     4261 gcgcagcaac agatggaggg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     4321 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     4381 ctccccggag tggaggccgc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     4441 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     4501 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctaggc tagtatgtaa
     4561 gcctagtctt agataataaa atcgctatcc atcgaagatg gatgtgtgtt ggttttttgt
     4621 gtgtgtaacg ctaggcgcgc ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac
     4681 gatccatatc gttgcttggc agtcagccag tcggaatcca gcttgggacc caggaagtcc
     4741 aatcgtcaga tattgtactc aagcctggtc acggcagcgt accgatctgt ttaaacctag
     4801 atattgatag tctgatcggt caacgtataa tcgagtccta gcttttgcaa acatctatca
     4861 agagacagga tcagcaggag gctttcgcat gagtattcaa catttccgtg tcgcccttat
     4921 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4981 aaaagatgct gaagatcagt tgggtgcgcg agtgggttac atcgaactgg atctcaacag
     5041 cggtaagatc cttgagagtt ttcgccccga agaacgcttt ccaatgatga gcacttttaa
     5101 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     5161 ccgcatacac tattctcaga atgacttggt tgagtattca ccagtcacag aaaagcatct
     5221 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5281 tgcggccaac ttacttctga caacgattgg aggaccgaag gagctaaccg cttttttgca
     5341 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5401 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact
     5461 attaactggc gaactactta ctctagcttc ccggcaacag ttgatagact ggatggaggc
     5521 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5581 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5641 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5701 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac cgattctagg
     5761 tgcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg
     5821 ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta
     5881 ccagaaattt atccttaaga tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt
     5941 ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga atctcgtcag
     6001 ctatcgtcag cttacctttt tggcagcgat cgcggctccc gacatcttgg accattagct
     6061 ccacaggtat cttcttccct ctagtggtca taacagcagc ttcagctacc tctcaattca
     6121 aaaaacccct caagacccgt ttagaggccc caaggggtta tgctatcaat cgttgcgtta
     6181 cacacacaaa aaaccaacac acatccatct tcgatggata gcgattttat tatctaactg
     6241 ctgatcgagt gtagccagat ctagtaatca attacggggt cattagttca tagcccatat
     6301 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     6361 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     6421 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     6481 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     6541 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     6601 atcgctatta ccatgctgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
     6661 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     6721 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
     6781 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc
     6841 agatctttgt cgatcctacc atccactcga cacacccgcc agcggccgc
//
