LOCUS       sequence-307308-        6419 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 6419)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6419)
  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-m4252872
FEATURES             Location/Qualifiers
     source          1..6419
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    35..106
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(111..157)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    175..223
                     /note="synthetic polyadenylation signal"
     enhancer        304..607
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        609..811
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     762..782
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1111..1131
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1132..1209
                     /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"
     polyA_signal    1262..1383
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1273..1292)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1327..1346
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1498..1544
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1574..1710
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1755..1826
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2052..2640)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2132..2151)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3140..4349
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4186..4205
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4353..4952
                     /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(4353..4372)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4849..4869
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4979..5027
                     /note="synthetic polyadenylation signal"
     CDS             join(5293..5361,5362..6153)
                     /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(5511..5530)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6166..6302
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 agcttacctt tttggcagcg atcgcggctc ccgacatctt ggaccattag ctccacaggt
       61 atcttcttcc ctctagtggt cataacagca gcttcagcta cctctcaatt caaaaaaccc
      121 ctcaagaccc gtttagaggc cccaaggggt tatgctatca atcgttgcgt tacacacaca
      181 aaaaaccaac acacatccat cttcgatgga tagcgatttt attatctaac tgctgatcga
      241 gtgtagccag atctagtaat caattacggg gtcattagtt catagcccat atatggagtt
      301 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      361 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      421 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      481 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      541 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      601 taccatgctg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      661 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      721 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      781 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tcagatcttt
      841 gtcgatccta ccatccactc gacacacccg ccagcggccg ccaccatgta ctcattcgtt
      901 tcggaagaga caggtacgtt aatagttaat agcgtacttc tttttcttgc tttcgtggta
      961 ttcttgctag ttacactagc catccttact gcgcttcgat tgtgtgcgta ctgctgcaat
     1021 attgttaacg tgagtcttgt aaaaccttct ttttacgttt actctcgtgt taaaaatctg
     1081 aattcttcta gagttcctga tcttctggtc gaaaaccttt actttcagtc gaagaggcga
     1141 tggaaaaaga attttatcgc ggtgagtgca gccaacagat tcaaaaagat atcctcgtct
     1201 ggtgccctct aagctagctt gactgactga gatacagcgt accttcagct cacagacatg
     1261 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     1321 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     1381 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     1441 ttttaaagca agtaaaacct ctacaaatgt ggtattggcc catctctatc ggtatcgtag
     1501 cataacccct tggggcctct aaacgggtct tgaggggttt tttgtgcccc tcgggccgga
     1561 ttgctatcta ccggcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata
     1621 ctcccacata tgccagattc agcaacggat acggcttccc caacttgccc acttccatac
     1681 gtgtcctcct taccagaaat ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc
     1741 gatttcgatc aagacattcc tttaatggtc ttttctggac accactaggg gtcagaagta
     1801 gttcatcaaa ctttcttccc tccctaatct cattggttac cttgggctat cgaaacttaa
     1861 ttaaccagtc aagtcagcta cttggcgaga tcgacttgtc tgggtttcga ctacgctcag
     1921 aattgcgtca gtcaagttcg atctggtcct tgctattgca cccgttctcc gattacgagt
     1981 ttcatttaaa tcatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2041 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     2101 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     2161 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     2221 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     2281 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     2341 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     2401 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     2461 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     2521 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     2581 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     2641 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     2701 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     2761 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     2821 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcattt
     2881 aaatttccga actctccaag gccctcgtcg gaaaatcttc aaacctttcg tccgatccat
     2941 cttgcaggct acctctcgaa cgaactatcg caagtctctt ggccggcctt gcgccttggc
     3001 tattgcttgg cagcgcctat cgccaggtat tactccaatc ccgaatatcc gagatcggga
     3061 tcacccgaga gaagttcaac ctacatcctc aatcccgatc tatccgagat ccgaggaata
     3121 tcgaaatcgg ggcgcgcctg gcctccgcgc cgggttttgg cgcctcccgc gggcgccccc
     3181 ctcgtcacgg cgagcgctgc cacgtcagac gaagggcgca ggagcgtcct gatccttccg
     3241 cccggacgct caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc
     3301 agcagaagga cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag
     3361 agagcggaac aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt
     3421 ggggcggtga acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt
     3481 cgcagccggg atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga
     3541 gtagcgggct gctgggctgg ccggggcttt cgtggccgcc gggccgctcg gtgggacgga
     3601 agcgtgtgga gagaccgcca agggctgtag tctgggtccg cgagcaaggt tgccctgaac
     3661 tgggggttgg ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc
     3721 ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg cggcaagaac
     3781 ccaaggtctt gagcccttcg ctaatgcggg aaagctctta ttcgggtgag atgggctggg
     3841 caccatctgg ggaccctgac gtgaagtttg tcactgactg gagaactcgg tttgtcgtct
     3901 gttgcggggg cggcagttat ggcggtgccg ttgggcagtg cacccgtacc tttgggagcg
     3961 cgcgccctcg tcgtgtcgtg acgtcacccg ttctgttggc ttataatgca gggtggggcc
     4021 accagccggt aggtgtgcgg taggcttttc tccgtcgcag gacgcagggt tcgggcctag
     4081 ggtaggctct cctgaatcga caggcgccgg acctctggtg aggggaggga taagtgaggc
     4141 gtcagtttct ttggtcggtt ttatgtacct atcttcttaa gtagctgaag ctccggtttt
     4201 gaactatgcg ctcggggttg gcgagtgtgt tttgtgaagt tttttaggca ccttttgaaa
     4261 tgtaatcatt tgggtcaata tgtaattttc agtgttagac ttgtaaattg tccgctaaat
     4321 tctggccgtt tttggctttt ttgttagaca acatgaccga gtacaagccc acggtgcgcc
     4381 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact
     4441 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc
     4501 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     4561 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     4621 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     4681 agggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     4741 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     4801 ccgccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     4861 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     4921 cctggtgcat gacccgcaag cccggtgcct aggctagtat gtaagcctag tcttagataa
     4981 taaaatcgct atccatcgaa gatggatgtg tgttggtttt ttgtgtgtgt aacgctaggc
     5041 gcgcctggtg taccgagaac gatcctctca gtgcgagtct cgacgatcca tatcgttgct
     5101 tggcagtcag ccagtcggaa tccagcttgg gacccaggaa gtccaatcgt cagatattgt
     5161 actcaagcct ggtcacggca gcgtaccgat ctgtttaaac ctagatattg atagtctgat
     5221 cggtcaacgt ataatcgagt cctagctttt gcaaacatct atcaagagac aggatcagca
     5281 ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5341 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5401 cagttgggtg cgcgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5461 agttttcgcc ccgaagaacg ctttccaatg atgagcactt ttaaagttct gctatgtggc
     5521 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5581 cagaatgact tggttgagta ttcaccagtc acagaaaagc atcttacgga tggcatgaca
     5641 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5701 ctgacaacga ttggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5761 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5821 gacaccacga tgcctgtagc aatggcaaca accttgcgta aactattaac tggcgaacta
     5881 cttactctag cttcccggca acagttgata gactggatgg aggcggataa agttgcagga
     5941 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6001 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6061 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6121 gagataggtg cctcactgat taagcattgg taaccgattc taggtgcatt ggcgcagaaa
     6181 aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg
     6241 atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt
     6301 aagatcccga atcgtttaaa ctcgactctg gctctatcga atctccgtcg tttcgagctt
     6361 acgcgaacag ccgtggcgct catttgctcg tcgggcatcg aatctcgtca gctatcgtc
//
