LOCUS       sequence-300439-        6765 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 SARS-CoV-2 E (envelope).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6765)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6765)
  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-m4252984
FEATURES             Location/Qualifiers
     source          1..6765
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      45..91
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    96..167
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(319..455)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(468..1259,1260..1328))
                     /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     1091..1110
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1594..1642
                     /note="synthetic polyadenylation signal"
     CDS             complement(1669..2697)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(2701..3910)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(2845..2864)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      4410..4998
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4899..4918
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5224..5295
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5340..5476)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(5506..5552)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5667..5788
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5704..5723)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5758..5777
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        complement(6156..6358)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6185..6205)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        6360..6663
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    join(6744..6765,1..27)
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 gatggatgtg tgttggtttt ttgtgtgtgt aacgcaacga ttgatagcat aaccccttgg
       61 ggcctctaaa cgggtcttga ggggtttttt gaattgagag gtagctgaag ctgctgttat
      121 gaccactaga gggaagaaga tacctgtgga gctaatggtc caagatgtcg ggagccgcga
      181 tcgctgccaa aaaggtaagc tgacgatagc tgacgagatt cgatgcccga cgagcaaatg
      241 agcgccacgg ctgttcgcgt aagctcgaaa cgacggagat tcgatagagc cagagtcgag
      301 tttaaacgat tcgggatctt aaggataaat ttctggtaag gaggacacgt atggaagtgg
      361 gcaagttggg gaagccgtat ccgttgctga atctggcata tgtgggagta taagacgcgc
      421 agcgtcgcat caggcatttt tttctgcgcc aatgcaccta gaatcggtta ccaatgctta
      481 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
      541 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
      601 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
      661 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tatcaactgt
      721 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tacgcaaggt tgttgccatt
      781 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
      841 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
      901 ggtcctccaa tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
      961 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgaa
     1021 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     1081 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaag
     1141 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     1201 cccactcgcg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     1261 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     1321 atactcatgc gaaagcctcc tgctgatcct gtctcttgat agatgtttgc aaaagctagg
     1381 actcgattat acgttgaccg atcagactat caatatctag gtttaaacag atcggtacgc
     1441 tgccgtgacc aggcttgagt acaatatctg acgattggac ttcctgggtc ccaagctgga
     1501 ttccgactgg ctgactgcca agcaacgata tggatcgtcg agactcgcac tgagaggatc
     1561 gttctcggta caccaggcgc gcctagcgtt acacacacaa aaaaccaaca cacatccatc
     1621 ttcgatggat agcgatttta ttatctaaga ctaggcttac atactagctt attcctttgc
     1681 cctcggacga gtgctggggc gtcggtttcc actatcggcg agtacttcta cacagccatc
     1741 ggtccagacg gcggcgcttc tgcgggcgat ttgtgtacgc ccgacagtcc cggctccgga
     1801 tcggacgatt gcgtcgcatc gaccctgcgc ccaagctgca tcatcgaaat tgccgtcaac
     1861 caagctctga tagagttggt caagaccaat gcggagcata tacgcccgga gccggggcga
     1921 tcctgcaagc tccggatgcc tccgctcgaa gtagcgcgtc tgctgctcca tacaagccaa
     1981 ccacggcctc cagaagaaga tgttggcgac ctcgtattgg gaatccccga acatcgcctc
     2041 gctccagtca atgaccgctg ttatgcggcc attgtccgtc aggacattgt tggagccgaa
     2101 atccgcgtgc acgaggtgcc ggacttcggg gcagtcctcg gcccaaagca tcagctcatc
     2161 gagagcctgc gcgacggacg cactgacggt gtcgtccatc acagtttgcc agtgatacac
     2221 atggggatca gcaatcgcgc atatgaaatc acgccatgta gtgtattgac cgattccttg
     2281 cggtccgaat gggccgaacc cgctcgtctg gctaagatcg gcggcagcga tggcatccat
     2341 tgcctccgcg accggctgca gaacagcggg cagttcggtt tcaggcaggt cttgcaacgt
     2401 gacaccctgt gcacggcggg agatgcaata ggtcaggctc tcgctgaact ccccaatgtc
     2461 aagcacttcc ggaatcggga gcgcggcgga tgcaaagtgc cgataaacat aacgatcttt
     2521 gtagaaacca tcggcgcagc tatttacccg caggacatat ccacgccctc ctacatcgaa
     2581 gctgaaagca cgagattctt cgccctccga gagctgcatc aggtcggaga cgctgtcgaa
     2641 cttttcgatc agaaacttct cgacagacgt cgcggtgagt tcaggctttt tacccatgtt
     2701 gtctaacaaa aaagccaaaa acggccagaa tttagcggac aatttacaag tctaacactg
     2761 aaaattacat attgacccaa atgattacat ttcaaaaggt gcctaaaaaa cttcacaaaa
     2821 cacactcgcc aaccccgagc gcatagttca aaaccggagc ttcagctact taagaagata
     2881 ggtacataaa accgaccaaa gaaactgacg cctcacttat ccctcccctc accagaggtc
     2941 cggcgcctgt cgattcagga gagcctaccc taggcccgaa ccctgcgtcc tgcgacggag
     3001 aaaagcctac cgcacaccta ccggctggtg gccccaccct gcattataag ccaacagaac
     3061 gggtgacgtc acgacacgac gagggcgcgc gctcccaaag gtacgggtgc actgcccaac
     3121 ggcaccgcca taactgccgc ccccgcaaca gacgacaaac cgagttctcc agtcagtgac
     3181 aaacttcacg tcagggtccc cagatggtgc ccagcccatc tcacccgaat aagagctttc
     3241 ccgcattagc gaagggctca agaccttggg ttcttgccgc ccaccatgcc ccccaccttg
     3301 tttcaacgac ctcacagccc gcctcacaag cgtcttccat tcaagactcg ggaacagccg
     3361 ccattttgct gcgctccccc caacccccag ttcagggcaa ccttgctcgc ggacccagac
     3421 tacagccctt ggcggtctct ccacacgctt ccgtcccacc gagcggcccg gcggccacga
     3481 aagccccggc cagcccagca gcccgctact caccaagtga cgatcacagc gatccacaaa
     3541 caagaaccgc gacccaaatc ccggctgcga cggaactagc tgtgccacac ccggcgcgtc
     3601 cttatataat catcggcgtt caccgcccca cggagatccc tccgcagaat cgccgagaag
     3661 ggactacttt tcctcgcctg ttccgctctc tggaaagaaa accagtgccc tagagtcacc
     3721 caagtcccgt cctaaaatgt ccttctgctg atactggggt tctaaggccg agtcttatga
     3781 gcagcgggcc gctgtcctga gcgtccgggc ggaaggatca ggacgctcct gcgcccttcg
     3841 tctgacgtgg cagcgctcgc cgtgacgagg ggggcgcccg cgggaggcgc caaaacccgg
     3901 cgcggaggcc aggcgcgccc cgatttcgat attcctcgga tctcggatag atcgggattg
     3961 aggatgtagg ttgaacttct ctcgggtgat cccgatctcg gatattcggg attggagtaa
     4021 tacctggcga taggcgctgc caagcaatag ccaaggcgca aggccggcca agagacttgc
     4081 gatagttcgt tcgagaggta gcctgcaaga tggatcggac gaaaggtttg aagattttcc
     4141 gacgagggcc ttggagagtt cggaaattta aatgcaacta tggatgaacg aaatagacag
     4201 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     4261 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4321 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4381 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4441 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4501 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4561 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4621 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4681 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4741 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4801 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4861 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4921 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4981 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5041 tggccttttg ctcacatgat ttaaatgaaa ctcgtaatcg gagaacgggt gcaatagcaa
     5101 ggaccagatc gaacttgact gacgcaattc tgagcgtagt cgaaacccag acaagtcgat
     5161 ctcgccaagt agctgacttg actggttaat taagtttcga tagcccaagg taaccaatga
     5221 gattagggag ggaagaaagt ttgatgaact acttctgacc cctagtggtg tccagaaaag
     5281 accattaaag gaatgtcttg atcgaaatcg agagatcgcc tgcaggatag ctgacgacct
     5341 taaggataaa tttctggtaa ggaggacacg tatggaagtg ggcaagttgg ggaagccgta
     5401 tccgttgctg aatctggcat atgtgggagt ataagacgcg cagcgtcgca tcaggcattt
     5461 ttttctgcgc caatgccggt agatagcaat ccggcccgag gggcacaaaa aacccctcaa
     5521 gacccgttta gaggccccaa ggggttatgc tacgataccg atagagatgg gccaatacca
     5581 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     5641 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     5701 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     5761 gtttgtccaa actcatcaat gtatcttatc atgtctgtga gctgaaggta cgctgtatct
     5821 cagtcagtca agctagcaga ggcctgcgga tccttagacc agaagatcag gaactctaga
     5881 agaattcaga tttttaacac gagagtaaac gtaaaaagaa ggttttacaa gactcacgtt
     5941 aacaatattg cagcagtacg cacacaatcg aagcgcagta aggatggcta gtgtaactag
     6001 caagaatacc acgaaagcaa gaaaaagaag tacgctatta actattaacg tacctgtctc
     6061 ttccgaaacg aatgagtaca tggtggcggc cgctggcggg tgtgtcgagt ggatggtagg
     6121 atcgacaaag atctgatctg acggttcact aaaccagctc tgcttatata gacctcccac
     6181 cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag
     6241 tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg gagacttgga
     6301 aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa ccgcatcagc
     6361 atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc
     6421 atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta
     6481 cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc
     6541 cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac
     6601 gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta
     6661 acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta ctagatctgg
     6721 ctacactcga tcagcagtta gataataaaa tcgctatcca tcgaa
//
