LOCUS       sequence-300642-        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-m4252827
FEATURES             Location/Qualifiers
     source          1..6896
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(66..86)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(114..191)
                     /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(225..245)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(545..747)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(574..594)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        749..1052
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    1133..1181
                     /note="synthetic polyadenylation signal"
     terminator      1199..1245
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    1250..1321
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(1473..1609)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(1622..2413,2414..2482))
                     /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     2245..2264
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    2748..2796
                     /note="synthetic polyadenylation signal"
     CDS             complement(2823..3422)
                     /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(2906..2926)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     3403..3422
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(3426..4635)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(3570..3589)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      5135..5723
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5624..5643
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5949..6020
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6065..6201)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(6231..6277)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6392..6513
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6429..6448)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6483..6502
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 gcgttctgct gttctttatt aaacttgtta tcgacggcct cgtcatgttg tgccaagcca
       61 aattcaccct ggaaatagag attctcggag gcggtcgtag gaatgtcgta atctagggcg
      121 ccgcttgaac tgatcttttt gaaccggtta gccgcagaca ctgcaataaa gttcttcttc
      181 catctccttt tccagggatt tatcagcccc gggattcgcc gtcccgactg aaagtaaagg
      241 ttttcgacca gaagatcagg aactctagaa gaattcagat ttttaacacg agagtaaacg
      301 taaaaagaag gttttacaag actcacgtta acaatattgc agcagtacgc acacaatcga
      361 agcgcagtaa ggatggctag tgtaactagc aagaatacca cgaaagcaag aaaaagaagt
      421 acgctattaa ctattaacgt acctgtctct tccgaaacga atgagtacat ggtggcggcc
      481 gctggcgggt gtgtcgagtg gatggtagga tcgacaaaga tctgatctga cggttcacta
      541 aaccagctct gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg
      601 gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc
      661 cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc
      721 cattgatgta ctgccaaaac cgcatcagca tggtaatagc gatgactaat acgtagatgt
      781 actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt
      841 taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca
      901 agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg
      961 ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc
     1021 caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta
     1081 atgaccccgt aattgattac tagatctggc tacactcgat cagcagttag ataataaaat
     1141 cgctatccat cgaagatgga tgtgtgttgg ttttttgtgt gtgtaacgca acgattgata
     1201 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgaattg agaggtagct
     1261 gaagctgctg ttatgaccac tagagggaag aagatacctg tggagctaat ggtccaagat
     1321 gtcgggagcc gcgatcgctg ccaaaaaggt aagctgacga tagctgacga gattcgatgc
     1381 ccgacgagca aatgagcgcc acggctgttc gcgtaagctc gaaacgacgg agattcgata
     1441 gagccagagt cgagtttaaa cgattcggga tcttaaggat aaatttctgg taaggaggac
     1501 acgtatggaa gtgggcaagt tggggaagcc gtatccgttg ctgaatctgg catatgtggg
     1561 agtataagac gcgcagcgtc gcatcaggca tttttttctg cgccaatgca cctagaatcg
     1621 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     1681 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     1741 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     1801 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     1861 agtctatcaa ctgttgccgg gaagctagag taagtagttc gccagttaat agtttacgca
     1921 aggttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     1981 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     2041 cggttagctc cttcggtcct ccaatcgttg tcagaagtaa gttggccgca gtgttatcac
     2101 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     2161 ctgtgactgg tgaatactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     2221 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     2281 tcatcattgg aaagcgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     2341 ccagttcgat gtaacccact cgcgcaccca actgatcttc agcatctttt actttcacca
     2401 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     2461 cacggaaatg ttgaatactc atgcgaaagc ctcctgctga tcctgtctct tgatagatgt
     2521 ttgcaaaagc taggactcga ttatacgttg accgatcaga ctatcaatat ctaggtttaa
     2581 acagatcggt acgctgccgt gaccaggctt gagtacaata tctgacgatt ggacttcctg
     2641 ggtcccaagc tggattccga ctggctgact gccaagcaac gatatggatc gtcgagactc
     2701 gcactgagag gatcgttctc ggtacaccag gcgcgcctag cgttacacac acaaaaaacc
     2761 aacacacatc catcttcgat ggatagcgat tttattatct aagactaggc ttacatacta
     2821 gcctaggcac cgggcttgcg ggtcatgcac caggtgcgcg gtccttcggg cacctcgacg
     2881 tcggcggtga cggtgaagcc gagccgctcg tagaagggga ggttgcgggg cgcggaggtc
     2941 tccaggaagg cgggcacccc ggcgcgctcg gcggcctcca ctccggggag cacgacggcg
     3001 ctgcccagac ccttgccctg gtggtcgggc gagacgccga cggtggccag gaaccacgcg
     3061 ggctccttgg gccggtgcgg cgccaggagg ccctccatct gttgctgcgc ggccagccgg
     3121 gaaccgctca actcggccat gcgcgggccg atctcggcga acaccgcccc cgcttcgacg
     3181 ctctccggcg tggtccagac cgccaccgcg gcgccgtcgt ccgcgaccca caccttgccg
     3241 atgtcgagcc cgacgcgcgt gaggaagagt tcttgcagct cggtgacccg ctcgatgtgg
     3301 cggtccggat cgacggtgtg gcgcgtggcg gggtagtcgg cgaacgcggc ggcgagggtg
     3361 cgtacggccc tggggacgtc gtcgcgggtg gcgaggcgca ccgtgggctt gtactcggtc
     3421 atgttgtcta acaaaaaagc caaaaacggc cagaatttag cggacaattt acaagtctaa
     3481 cactgaaaat tacatattga cccaaatgat tacatttcaa aaggtgccta aaaaacttca
     3541 caaaacacac tcgccaaccc cgagcgcata gttcaaaacc ggagcttcag ctacttaaga
     3601 agataggtac ataaaaccga ccaaagaaac tgacgcctca cttatccctc ccctcaccag
     3661 aggtccggcg cctgtcgatt caggagagcc taccctaggc ccgaaccctg cgtcctgcga
     3721 cggagaaaag cctaccgcac acctaccggc tggtggcccc accctgcatt ataagccaac
     3781 agaacgggtg acgtcacgac acgacgaggg cgcgcgctcc caaaggtacg ggtgcactgc
     3841 ccaacggcac cgccataact gccgcccccg caacagacga caaaccgagt tctccagtca
     3901 gtgacaaact tcacgtcagg gtccccagat ggtgcccagc ccatctcacc cgaataagag
     3961 ctttcccgca ttagcgaagg gctcaagacc ttgggttctt gccgcccacc atgcccccca
     4021 ccttgtttca acgacctcac agcccgcctc acaagcgtct tccattcaag actcgggaac
     4081 agccgccatt ttgctgcgct ccccccaacc cccagttcag ggcaaccttg ctcgcggacc
     4141 cagactacag cccttggcgg tctctccaca cgcttccgtc ccaccgagcg gcccggcggc
     4201 cacgaaagcc ccggccagcc cagcagcccg ctactcacca agtgacgatc acagcgatcc
     4261 acaaacaaga accgcgaccc aaatcccggc tgcgacggaa ctagctgtgc cacacccggc
     4321 gcgtccttat ataatcatcg gcgttcaccg ccccacggag atccctccgc agaatcgccg
     4381 agaagggact acttttcctc gcctgttccg ctctctggaa agaaaaccag tgccctagag
     4441 tcacccaagt cccgtcctaa aatgtccttc tgctgatact ggggttctaa ggccgagtct
     4501 tatgagcagc gggccgctgt cctgagcgtc cgggcggaag gatcaggacg ctcctgcgcc
     4561 cttcgtctga cgtggcagcg ctcgccgtga cgaggggggc gcccgcggga ggcgccaaaa
     4621 cccggcgcgg aggccaggcg cgccccgatt tcgatattcc tcggatctcg gatagatcgg
     4681 gattgaggat gtaggttgaa cttctctcgg gtgatcccga tctcggatat tcgggattgg
     4741 agtaatacct ggcgataggc gctgccaagc aatagccaag gcgcaaggcc ggccaagaga
     4801 cttgcgatag ttcgttcgag aggtagcctg caagatggat cggacgaaag gtttgaagat
     4861 tttccgacga gggccttgga gagttcggaa atttaaatgc aactatggat gaacgaaata
     4921 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4981 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     5041 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5101 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5161 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5221 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5281 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5341 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5401 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5461 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5521 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5581 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5641 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5701 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5761 tttgctggcc ttttgctcac atgatttaaa tgaaactcgt aatcggagaa cgggtgcaat
     5821 agcaaggacc agatcgaact tgactgacgc aattctgagc gtagtcgaaa cccagacaag
     5881 tcgatctcgc caagtagctg acttgactgg ttaattaagt ttcgatagcc caaggtaacc
     5941 aatgagatta gggagggaag aaagtttgat gaactacttc tgacccctag tggtgtccag
     6001 aaaagaccat taaaggaatg tcttgatcga aatcgagaga tcgcctgcag gatagctgac
     6061 gaccttaagg ataaatttct ggtaaggagg acacgtatgg aagtgggcaa gttggggaag
     6121 ccgtatccgt tgctgaatct ggcatatgtg ggagtataag acgcgcagcg tcgcatcagg
     6181 catttttttc tgcgccaatg ccggtagata gcaatccggc ccgaggggca caaaaaaccc
     6241 ctcaagaccc gtttagaggc cccaaggggt tatgctacga taccgataga gatgggccaa
     6301 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     6361 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     6421 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     6481 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtgagctga aggtacgctg
     6541 tatctcagtc agtcaagcta gcttattgat ggttcgcgtc tactttcggg gcctgggcat
     6601 cattgagctt tttggcttcc gcaagcaggt tggcactctg tgatggatcg tcttttaggg
     6661 actggataaa tgcatttctt tgttcctcat tcagattagg aaggtgcaat atctcgtaga
     6721 aagcattctg ctgctccttg ttaaacttgt tatcaacttt tggagcctgt gcgtcattca
     6781 gcttcttggc ttcggccagg agatttgcag actgcgaggg atcgtctttc aggctctgaa
     6841 tgaaagcgtt gcgttgctct tcgttcagat ttggtaagtg aaggatctca tagaaa
//
