LOCUS       sequence-301664-        7341 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 7341)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7341)
  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-m4252907
FEATURES             Location/Qualifiers
     source          1..7341
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    19..67
                     /note="synthetic polyadenylation signal"
     terminator      85..131
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    136..207
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(359..495)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(508..1299,1300..1368))
                     /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     1131..1150
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1634..1682
                     /note="synthetic polyadenylation signal"
     CDS             complement(1709..2737)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(2741..3950)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(2885..2904)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      4450..5038
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4939..4958
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5264..5335
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5380..5516)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(5546..5592)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5707..5828
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5744..5763)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5798..5817
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(6293..6313)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(6341..6418)
                     /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(6452..6472)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(6772..6974)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6801..6821)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6976..7279
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ctcgatcagc agttagataa taaaatcgct atccatcgaa gatggatgtg tgttggtttt
       61 ttgtgtgtgt aacgcaacga ttgatagcat aaccccttgg ggcctctaaa cgggtcttga
      121 ggggtttttt gaattgagag gtagctgaag ctgctgttat gaccactaga gggaagaaga
      181 tacctgtgga gctaatggtc caagatgtcg ggagccgcga tcgctgccaa aaaggtaagc
      241 tgacgatagc tgacgagatt cgatgcccga cgagcaaatg agcgccacgg ctgttcgcgt
      301 aagctcgaaa cgacggagat tcgatagagc cagagtcgag tttaaacgat tcgggatctt
      361 aaggataaat ttctggtaag gaggacacgt atggaagtgg gcaagttggg gaagccgtat
      421 ccgttgctga atctggcata tgtgggagta taagacgcgc agcgtcgcat caggcatttt
      481 tttctgcgcc aatgcaccta gaatcggtta ccaatgctta atcagtgagg cacctatctc
      541 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
      601 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
      661 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
      721 tcctgcaact ttatccgcct ccatccagtc tatcaactgt tgccgggaag ctagagtaag
      781 tagttcgcca gttaatagtt tacgcaaggt tgttgccatt gctacaggca tcgtggtgtc
      841 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
      901 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccaa tcgttgtcag
      961 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     1021 tgtcatgcca tccgtaagat gcttttctgt gactggtgaa tactcaacca agtcattctg
     1081 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     1141 gccacatagc agaactttaa aagtgctcat cattggaaag cgttcttcgg ggcgaaaact
     1201 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgcg cacccaactg
     1261 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     1321 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatgc gaaagcctcc
     1381 tgctgatcct gtctcttgat agatgtttgc aaaagctagg actcgattat acgttgaccg
     1441 atcagactat caatatctag gtttaaacag atcggtacgc tgccgtgacc aggcttgagt
     1501 acaatatctg acgattggac ttcctgggtc ccaagctgga ttccgactgg ctgactgcca
     1561 agcaacgata tggatcgtcg agactcgcac tgagaggatc gttctcggta caccaggcgc
     1621 gcctagcgtt acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta
     1681 ttatctaaga ctaggcttac atactagctt attcctttgc cctcggacga gtgctggggc
     1741 gtcggtttcc actatcggcg agtacttcta cacagccatc ggtccagacg gcggcgcttc
     1801 tgcgggcgat ttgtgtacgc ccgacagtcc cggctccgga tcggacgatt gcgtcgcatc
     1861 gaccctgcgc ccaagctgca tcatcgaaat tgccgtcaac caagctctga tagagttggt
     1921 caagaccaat gcggagcata tacgcccgga gccggggcga tcctgcaagc tccggatgcc
     1981 tccgctcgaa gtagcgcgtc tgctgctcca tacaagccaa ccacggcctc cagaagaaga
     2041 tgttggcgac ctcgtattgg gaatccccga acatcgcctc gctccagtca atgaccgctg
     2101 ttatgcggcc attgtccgtc aggacattgt tggagccgaa atccgcgtgc acgaggtgcc
     2161 ggacttcggg gcagtcctcg gcccaaagca tcagctcatc gagagcctgc gcgacggacg
     2221 cactgacggt gtcgtccatc acagtttgcc agtgatacac atggggatca gcaatcgcgc
     2281 atatgaaatc acgccatgta gtgtattgac cgattccttg cggtccgaat gggccgaacc
     2341 cgctcgtctg gctaagatcg gcggcagcga tggcatccat tgcctccgcg accggctgca
     2401 gaacagcggg cagttcggtt tcaggcaggt cttgcaacgt gacaccctgt gcacggcggg
     2461 agatgcaata ggtcaggctc tcgctgaact ccccaatgtc aagcacttcc ggaatcggga
     2521 gcgcggcgga tgcaaagtgc cgataaacat aacgatcttt gtagaaacca tcggcgcagc
     2581 tatttacccg caggacatat ccacgccctc ctacatcgaa gctgaaagca cgagattctt
     2641 cgccctccga gagctgcatc aggtcggaga cgctgtcgaa cttttcgatc agaaacttct
     2701 cgacagacgt cgcggtgagt tcaggctttt tacccatgtt gtctaacaaa aaagccaaaa
     2761 acggccagaa tttagcggac aatttacaag tctaacactg aaaattacat attgacccaa
     2821 atgattacat ttcaaaaggt gcctaaaaaa cttcacaaaa cacactcgcc aaccccgagc
     2881 gcatagttca aaaccggagc ttcagctact taagaagata ggtacataaa accgaccaaa
     2941 gaaactgacg cctcacttat ccctcccctc accagaggtc cggcgcctgt cgattcagga
     3001 gagcctaccc taggcccgaa ccctgcgtcc tgcgacggag aaaagcctac cgcacaccta
     3061 ccggctggtg gccccaccct gcattataag ccaacagaac gggtgacgtc acgacacgac
     3121 gagggcgcgc gctcccaaag gtacgggtgc actgcccaac ggcaccgcca taactgccgc
     3181 ccccgcaaca gacgacaaac cgagttctcc agtcagtgac aaacttcacg tcagggtccc
     3241 cagatggtgc ccagcccatc tcacccgaat aagagctttc ccgcattagc gaagggctca
     3301 agaccttggg ttcttgccgc ccaccatgcc ccccaccttg tttcaacgac ctcacagccc
     3361 gcctcacaag cgtcttccat tcaagactcg ggaacagccg ccattttgct gcgctccccc
     3421 caacccccag ttcagggcaa ccttgctcgc ggacccagac tacagccctt ggcggtctct
     3481 ccacacgctt ccgtcccacc gagcggcccg gcggccacga aagccccggc cagcccagca
     3541 gcccgctact caccaagtga cgatcacagc gatccacaaa caagaaccgc gacccaaatc
     3601 ccggctgcga cggaactagc tgtgccacac ccggcgcgtc cttatataat catcggcgtt
     3661 caccgcccca cggagatccc tccgcagaat cgccgagaag ggactacttt tcctcgcctg
     3721 ttccgctctc tggaaagaaa accagtgccc tagagtcacc caagtcccgt cctaaaatgt
     3781 ccttctgctg atactggggt tctaaggccg agtcttatga gcagcgggcc gctgtcctga
     3841 gcgtccgggc ggaaggatca ggacgctcct gcgcccttcg tctgacgtgg cagcgctcgc
     3901 cgtgacgagg ggggcgcccg cgggaggcgc caaaacccgg cgcggaggcc aggcgcgccc
     3961 cgatttcgat attcctcgga tctcggatag atcgggattg aggatgtagg ttgaacttct
     4021 ctcgggtgat cccgatctcg gatattcggg attggagtaa tacctggcga taggcgctgc
     4081 caagcaatag ccaaggcgca aggccggcca agagacttgc gatagttcgt tcgagaggta
     4141 gcctgcaaga tggatcggac gaaaggtttg aagattttcc gacgagggcc ttggagagtt
     4201 cggaaattta aatgcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4261 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     4321 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4381 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4441 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4501 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4561 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4621 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     4681 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     4741 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4801 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4861 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4921 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4981 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5041 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgat
     5101 ttaaatgaaa ctcgtaatcg gagaacgggt gcaatagcaa ggaccagatc gaacttgact
     5161 gacgcaattc tgagcgtagt cgaaacccag acaagtcgat ctcgccaagt agctgacttg
     5221 actggttaat taagtttcga tagcccaagg taaccaatga gattagggag ggaagaaagt
     5281 ttgatgaact acttctgacc cctagtggtg tccagaaaag accattaaag gaatgtcttg
     5341 atcgaaatcg agagatcgcc tgcaggatag ctgacgacct taaggataaa tttctggtaa
     5401 ggaggacacg tatggaagtg ggcaagttgg ggaagccgta tccgttgctg aatctggcat
     5461 atgtgggagt ataagacgcg cagcgtcgca tcaggcattt ttttctgcgc caatgccggt
     5521 agatagcaat ccggcccgag gggcacaaaa aacccctcaa gacccgttta gaggccccaa
     5581 ggggttatgc tacgataccg atagagatgg gccaatacca catttgtaga ggttttactt
     5641 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     5701 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     5761 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     5821 gtatcttatc atgtctgtga gctgaaggta cgctgtatct cagtcagtca agctagcaga
     5881 ggcctgcgga tccttattga tggttcgcgt ctactttcgg ggcctgggca tcattgagct
     5941 ttttggcttc cgcaagcagg ttggcactct gtgatggatc gtcttttagg gactggataa
     6001 atgcatttct ttgttcctca ttcagattag gaaggtgcaa tatctcgtag aaagcattct
     6061 gctgctcctt gttaaacttg ttatcaactt ttggagcctg tgcgtcattc agcttcttgg
     6121 cttcggccag gagatttgca gactgcgagg gatcgtcttt caggctctga atgaaagcgt
     6181 tgcgttgctc ttcgttcaga tttggtaagt gaaggatctc atagaaagcg ttctgctgtt
     6241 ctttattaaa cttgttatcg acggcctcgt catgttgtgc caagccaaat tcaccctgga
     6301 aatagagatt ctcggaggcg gtcgtaggaa tgtcgtaatc tagggcgccg cttgaactga
     6361 tctttttgaa ccggttagcc gcagacactg caataaagtt cttcttccat ctccttttcc
     6421 agggatttat cagccccggg attcgccgtc ccgactgaaa gtaaaggttt tcgaccagaa
     6481 gatcaggaac tctagaagaa ttcagatttt taacacgaga gtaaacgtaa aaagaaggtt
     6541 ttacaagact cacgttaaca atattgcagc agtacgcaca caatcgaagc gcagtaagga
     6601 tggctagtgt aactagcaag aataccacga aagcaagaaa aagaagtacg ctattaacta
     6661 ttaacgtacc tgtctcttcc gaaacgaatg agtacatggt ggcggccgct ggcgggtgtg
     6721 tcgagtggat ggtaggatcg acaaagatct gatctgacgg ttcactaaac cagctctgct
     6781 tatatagacc tcccaccgta cacgcctacc gcccatttgc gtcaatgggg cggagttgtt
     6841 acgacatttt ggaaagtccc gttgattttg gtgccaaaac aaactcccat tgacgtcaat
     6901 ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tgatgtactg
     6961 ccaaaaccgc atcagcatgg taatagcgat gactaatacg tagatgtact gccaagtagg
     7021 aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac
     7081 gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta
     7141 ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac
     7201 atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt
     7261 taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat
     7321 tgattactag atctggctac a
//
