LOCUS       sequence-297599-        6765 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 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-m4252989
FEATURES             Location/Qualifiers
     source          1..6765
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    298..419
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(309..328)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     363..382
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      534..580
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      610..746
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    791..862
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1088..1676)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1168..1187)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2176..3385
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3222..3241
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3389..4417
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4444..4492
                     /note="synthetic polyadenylation signal"
     CDS             join(4758..4826,4827..5618)
                     /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(4976..4995)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5631..5767
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5919..5990
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5995..6041)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6059..6107
                     /note="synthetic polyadenylation signal"
     enhancer        6188..6491
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6493..6695
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6646..6666
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 caccatgtac agctttgtat ctgaagagac aggcactctc atcgttaatt cagtgttgct
       61 gttcctggca tttgtggtct tccttttagt gaccctagcg atactgacgg ccctccggct
      121 gtgcgcttat tgttgtaaca ttgtcaacgt tagtctcgtg aagccctcgt tctacgttta
      181 ttcccgcgtg aaaaatctaa acagctcccg tgtgcctgat cttcttgtct aaggatccgc
      241 aggcctctgc tagcttgact gactgagata cagcgtacct tcagctcaca gacatgataa
      301 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
      361 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
      421 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
      481 aaagcaagta aaacctctac aaatgtggta ttggcccatc tctatcggta tcgtagcata
      541 accccttggg gcctctaaac gggtcttgag gggttttttg tgcccctcgg gccggattgc
      601 tatctaccgg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc
      661 cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt
      721 cctccttacc agaaatttat ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt
      781 tcgatcaaga cattccttta atggtctttt ctggacacca ctaggggtca gaagtagttc
      841 atcaaacttt cttccctccc taatctcatt ggttaccttg ggctatcgaa acttaattaa
      901 ccagtcaagt cagctacttg gcgagatcga cttgtctggg tttcgactac gctcagaatt
      961 gcgtcagtca agttcgatct ggtccttgct attgcacccg ttctccgatt acgagtttca
     1021 tttaaatcat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     1081 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     1141 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     1201 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     1261 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     1321 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     1381 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     1441 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     1501 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     1561 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     1621 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     1681 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     1741 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     1801 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     1861 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcatttaaat
     1921 ttccgaactc tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg
     1981 caggctacct ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc cttggctatt
     2041 gcttggcagc gcctatcgcc aggtattact ccaatcccga atatccgaga tcgggatcac
     2101 ccgagagaag ttcaacctac atcctcaatc ccgatctatc cgagatccga ggaatatcga
     2161 aatcggggcg cgcctggcct ccgcgccggg ttttggcgcc tcccgcgggc gcccccctcg
     2221 tcacggcgag cgctgccacg tcagacgaag ggcgcaggag cgtcctgatc cttccgcccg
     2281 gacgctcagg acagcggccc gctgctcata agactcggcc ttagaacccc agtatcagca
     2341 gaaggacatt ttaggacggg acttgggtga ctctagggca ctggttttct ttccagagag
     2401 cggaacaggc gaggaaaagt agtcccttct cggcgattct gcggagggat ctccgtgggg
     2461 cggtgaacgc cgatgattat ataaggacgc gccgggtgtg gcacagctag ttccgtcgca
     2521 gccgggattt gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag
     2581 cgggctgctg ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg gacggaagcg
     2641 tgtggagaga ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc ctgaactggg
     2701 ggttgggggg agcgcagcaa aatggcggct gttcccgagt cttgaatgga agacgcttgt
     2761 gaggcgggct gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc aagaacccaa
     2821 ggtcttgagc ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg gctgggcacc
     2881 atctggggac cctgacgtga agtttgtcac tgactggaga actcggtttg tcgtctgttg
     2941 cgggggcggc agttatggcg gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg
     3001 ccctcgtcgt gtcgtgacgt cacccgttct gttggcttat aatgcagggt ggggccacca
     3061 gccggtaggt gtgcggtagg cttttctccg tcgcaggacg cagggttcgg gcctagggta
     3121 ggctctcctg aatcgacagg cgccggacct ctggtgaggg gagggataag tgaggcgtca
     3181 gtttctttgg tcggttttat gtacctatct tcttaagtag ctgaagctcc ggttttgaac
     3241 tatgcgctcg gggttggcga gtgtgttttg tgaagttttt taggcacctt ttgaaatgta
     3301 atcatttggg tcaatatgta attttcagtg ttagacttgt aaattgtccg ctaaattctg
     3361 gccgtttttg gcttttttgt tagacaacat gggtaaaaag cctgaactca ccgcgacgtc
     3421 tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga
     3481 gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt
     3541 aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatccgc
     3601 cgcgctcccg attccggaag tgcttgacat tggggagttc agcgagagcc tgacctattg
     3661 catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc
     3721 tgttctgcag ccggtcgcgg aggcaatgga tgccatcgct gccgccgatc ttagccagac
     3781 gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt
     3841 catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt
     3901 cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga
     3961 agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg
     4021 cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc
     4081 caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga
     4141 gcggaggcat ccggagcttg caggatcgcc ccggctccgg gcgtatatgc tccgcattgg
     4201 tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca
     4261 gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc
     4321 ccgcagaagc gccgccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa
     4381 ccgacgcccc agcactcgtc cgagggcaaa ggaataagct agtatgtaag cctagtctta
     4441 gataataaaa tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg tgtgtaacgc
     4501 taggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg
     4561 ttgcttggca gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat
     4621 attgtactca agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt
     4681 ctgatcggtc aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat
     4741 cagcaggagg ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4801 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4861 aagatcagtt gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4921 ttgagagttt tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat
     4981 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5041 attctcagaa tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca
     5101 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5161 tacttctgac aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5221 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5281 agcgtgacac cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg
     5341 aactacttac tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg
     5401 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5461 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5521 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5581 tcgctgagat aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc
     5641 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
     5701 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
     5761 tccttaagat cccgaatcgt ttaaactcga ctctggctct atcgaatctc cgtcgtttcg
     5821 agcttacgcg aacagccgtg gcgctcattt gctcgtcggg catcgaatct cgtcagctat
     5881 cgtcagctta cctttttggc agcgatcgcg gctcccgaca tcttggacca ttagctccac
     5941 aggtatcttc ttccctctag tggtcataac agcagcttca gctacctctc aattcaaaaa
     6001 acccctcaag acccgtttag aggccccaag gggttatgct atcaatcgtt gcgttacaca
     6061 cacaaaaaac caacacacat ccatcttcga tggatagcga ttttattatc taactgctga
     6121 tcgagtgtag ccagatctag taatcaatta cggggtcatt agttcatagc ccatatatgg
     6181 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     6241 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     6301 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     6361 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     6421 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     6481 ctattaccat gctgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
     6541 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     6601 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
     6661 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatcagat
     6721 ctttgtcgat cctaccatcc actcgacaca cccgccagcg gccgc
//
