LOCUS       sequence-300602-        6896 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 6896)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6896)
  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-m4252868
FEATURES             Location/Qualifiers
     source          1..6896
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      249..837
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     738..757
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    1063..1134
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(1179..1315)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(1345..1391)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    1506..1627
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1543..1562)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1597..1616
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(2076..2096)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(2124..2201)
                     /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             complement(2235..2255)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(2555..2757)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2584..2604)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2759..3062
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    3143..3191
                     /note="synthetic polyadenylation signal"
     terminator      3209..3255
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    3260..3331
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3483..3619)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(3632..4423,4424..4492))
                     /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     4255..4274
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    4758..4806
                     /note="synthetic polyadenylation signal"
     CDS             complement(4833..5432)
                     /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(4916..4936)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     5413..5432
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(5436..6645)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(5580..5599)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
ORIGIN
        1 ggaaatttaa atgcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
       61 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
      121 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
      181 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
      241 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
      301 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
      361 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
      421 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
      481 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
      541 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
      601 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
      661 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
      721 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
      781 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
      841 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgatt
      901 taaatgaaac tcgtaatcgg agaacgggtg caatagcaag gaccagatcg aacttgactg
      961 acgcaattct gagcgtagtc gaaacccaga caagtcgatc tcgccaagta gctgacttga
     1021 ctggttaatt aagtttcgat agcccaaggt aaccaatgag attagggagg gaagaaagtt
     1081 tgatgaacta cttctgaccc ctagtggtgt ccagaaaaga ccattaaagg aatgtcttga
     1141 tcgaaatcga gagatcgcct gcaggatagc tgacgacctt aaggataaat ttctggtaag
     1201 gaggacacgt atggaagtgg gcaagttggg gaagccgtat ccgttgctga atctggcata
     1261 tgtgggagta taagacgcgc agcgtcgcat caggcatttt tttctgcgcc aatgccggta
     1321 gatagcaatc cggcccgagg ggcacaaaaa acccctcaag acccgtttag aggccccaag
     1381 gggttatgct acgataccga tagagatggg ccaataccac atttgtagag gttttacttg
     1441 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     1501 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     1561 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     1621 tatcttatca tgtctgtgag ctgaaggtac gctgtatctc agtcagtcaa gctagcttat
     1681 tgatggttcg cgtctacttt cggggcctgg gcatcattga gctttttggc ttccgcaagc
     1741 aggttggcac tctgtgatgg atcgtctttt agggactgga taaatgcatt tctttgttcc
     1801 tcattcagat taggaaggtg caatatctcg tagaaagcat tctgctgctc cttgttaaac
     1861 ttgttatcaa cttttggagc ctgtgcgtca ttcagcttct tggcttcggc caggagattt
     1921 gcagactgcg agggatcgtc tttcaggctc tgaatgaaag cgttgcgttg ctcttcgttc
     1981 agatttggta agtgaaggat ctcatagaaa gcgttctgct gttctttatt aaacttgtta
     2041 tcgacggcct cgtcatgttg tgccaagcca aattcaccct ggaaatagag attctcggag
     2101 gcggtcgtag gaatgtcgta atctagggcg ccgcttgaac tgatcttttt gaaccggtta
     2161 gccgcagaca ctgcaataaa gttcttcttc catctccttt tccagggatt tatcagcccc
     2221 gggattcgcc gtcccgactg aaagtaaagg ttttcgacaa gaagatcagg cacacgggag
     2281 ctgtttagat ttttcacgcg ggaataaacg tagaacgagg gcttcacgag actaacgttg
     2341 acaatgttac aacaataagc gcacagccgg agggccgtca gtatcgctag ggtcactaaa
     2401 aggaagacca caaatgccag gaacagcaac actgaattaa cgatgagagt gcctgtctct
     2461 tcagatacaa agctgtacat ggtggcggcc gctggcgggt gtgtcgagtg gatggtagga
     2521 tcgacaaaga tctgatctga cggttcacta aaccagctct gcttatatag acctcccacc
     2581 gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat tttggaaagt
     2641 cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg agacttggaa
     2701 atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcagca
     2761 tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca
     2821 tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac
     2881 ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc
     2941 attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg
     3001 tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa
     3061 cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac tagatctggc
     3121 tacactcgat cagcagttag ataataaaat cgctatccat cgaagatgga tgtgtgttgg
     3181 ttttttgtgt gtgtaacgca acgattgata gcataacccc ttggggcctc taaacgggtc
     3241 ttgaggggtt ttttgaattg agaggtagct gaagctgctg ttatgaccac tagagggaag
     3301 aagatacctg tggagctaat ggtccaagat gtcgggagcc gcgatcgctg ccaaaaaggt
     3361 aagctgacga tagctgacga gattcgatgc ccgacgagca aatgagcgcc acggctgttc
     3421 gcgtaagctc gaaacgacgg agattcgata gagccagagt cgagtttaaa cgattcggga
     3481 tcttaaggat aaatttctgg taaggaggac acgtatggaa gtgggcaagt tggggaagcc
     3541 gtatccgttg ctgaatctgg catatgtggg agtataagac gcgcagcgtc gcatcaggca
     3601 tttttttctg cgccaatgca cctagaatcg gttaccaatg cttaatcagt gaggcaccta
     3661 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3721 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3781 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3841 gtggtcctgc aactttatcc gcctccatcc agtctatcaa ctgttgccgg gaagctagag
     3901 taagtagttc gccagttaat agtttacgca aggttgttgc cattgctaca ggcatcgtgg
     3961 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4021 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccaatcgttg
     4081 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4141 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgaatactca accaagtcat
     4201 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4261 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaagcgttct tcggggcgaa
     4321 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgcgcaccca
     4381 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4441 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atgcgaaagc
     4501 ctcctgctga tcctgtctct tgatagatgt ttgcaaaagc taggactcga ttatacgttg
     4561 accgatcaga ctatcaatat ctaggtttaa acagatcggt acgctgccgt gaccaggctt
     4621 gagtacaata tctgacgatt ggacttcctg ggtcccaagc tggattccga ctggctgact
     4681 gccaagcaac gatatggatc gtcgagactc gcactgagag gatcgttctc ggtacaccag
     4741 gcgcgcctag cgttacacac acaaaaaacc aacacacatc catcttcgat ggatagcgat
     4801 tttattatct aagactaggc ttacatacta gcctaggcac cgggcttgcg ggtcatgcac
     4861 caggtgcgcg gtccttcggg cacctcgacg tcggcggtga cggtgaagcc gagccgctcg
     4921 tagaagggga ggttgcgggg cgcggaggtc tccaggaagg cgggcacccc ggcgcgctcg
     4981 gcggcctcca ctccggggag cacgacggcg ctgcccagac ccttgccctg gtggtcgggc
     5041 gagacgccga cggtggccag gaaccacgcg ggctccttgg gccggtgcgg cgccaggagg
     5101 ccctccatct gttgctgcgc ggccagccgg gaaccgctca actcggccat gcgcgggccg
     5161 atctcggcga acaccgcccc cgcttcgacg ctctccggcg tggtccagac cgccaccgcg
     5221 gcgccgtcgt ccgcgaccca caccttgccg atgtcgagcc cgacgcgcgt gaggaagagt
     5281 tcttgcagct cggtgacccg ctcgatgtgg cggtccggat cgacggtgtg gcgcgtggcg
     5341 gggtagtcgg cgaacgcggc ggcgagggtg cgtacggccc tggggacgtc gtcgcgggtg
     5401 gcgaggcgca ccgtgggctt gtactcggtc atgttgtcta acaaaaaagc caaaaacggc
     5461 cagaatttag cggacaattt acaagtctaa cactgaaaat tacatattga cccaaatgat
     5521 tacatttcaa aaggtgccta aaaaacttca caaaacacac tcgccaaccc cgagcgcata
     5581 gttcaaaacc ggagcttcag ctacttaaga agataggtac ataaaaccga ccaaagaaac
     5641 tgacgcctca cttatccctc ccctcaccag aggtccggcg cctgtcgatt caggagagcc
     5701 taccctaggc ccgaaccctg cgtcctgcga cggagaaaag cctaccgcac acctaccggc
     5761 tggtggcccc accctgcatt ataagccaac agaacgggtg acgtcacgac acgacgaggg
     5821 cgcgcgctcc caaaggtacg ggtgcactgc ccaacggcac cgccataact gccgcccccg
     5881 caacagacga caaaccgagt tctccagtca gtgacaaact tcacgtcagg gtccccagat
     5941 ggtgcccagc ccatctcacc cgaataagag ctttcccgca ttagcgaagg gctcaagacc
     6001 ttgggttctt gccgcccacc atgcccccca ccttgtttca acgacctcac agcccgcctc
     6061 acaagcgtct tccattcaag actcgggaac agccgccatt ttgctgcgct ccccccaacc
     6121 cccagttcag ggcaaccttg ctcgcggacc cagactacag cccttggcgg tctctccaca
     6181 cgcttccgtc ccaccgagcg gcccggcggc cacgaaagcc ccggccagcc cagcagcccg
     6241 ctactcacca agtgacgatc acagcgatcc acaaacaaga accgcgaccc aaatcccggc
     6301 tgcgacggaa ctagctgtgc cacacccggc gcgtccttat ataatcatcg gcgttcaccg
     6361 ccccacggag atccctccgc agaatcgccg agaagggact acttttcctc gcctgttccg
     6421 ctctctggaa agaaaaccag tgccctagag tcacccaagt cccgtcctaa aatgtccttc
     6481 tgctgatact ggggttctaa ggccgagtct tatgagcagc gggccgctgt cctgagcgtc
     6541 cgggcggaag gatcaggacg ctcctgcgcc cttcgtctga cgtggcagcg ctcgccgtga
     6601 cgaggggggc gcccgcggga ggcgccaaaa cccggcgcgg aggccaggcg cgccccgatt
     6661 tcgatattcc tcggatctcg gatagatcgg gattgaggat gtaggttgaa cttctctcgg
     6721 gtgatcccga tctcggatat tcgggattgg agtaatacct ggcgataggc gctgccaagc
     6781 aatagccaag gcgcaaggcc ggccaagaga cttgcgatag ttcgttcgag aggtagcctg
     6841 caagatggat cggacgaaag gtttgaagat tttccgacga gggccttgga gagttc
//
