LOCUS       sequence-300451-        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"
     misc_feature    44..115
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(120..166)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    184..232
                     /note="synthetic polyadenylation signal"
     enhancer        313..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        618..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    1188..1309
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1199..1218)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1253..1272
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1424..1470
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1500..1636
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1681..1752
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1978..2566)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2058..2077)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3066..4275
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4112..4131
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4279..5307
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5334..5382
                     /note="synthetic polyadenylation signal"
     CDS             join(5648..5716,5717..6508)
                     /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(5866..5885)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6521..6657
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg gaccattagc
       61 tccacaggta tcttcttccc tctagtggtc ataacagcag cttcagctac ctctcaattc
      121 aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt
      181 acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta ttatctaact
      241 gctgatcgag tgtagccaga tctagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatgctga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
      841 cagatctttg tcgatcctac catccactcg acacacccgc cagcggccgc caccatgtac
      901 agctttgtat ctgaagagac aggcactctc atcgttaatt cagtgttgct gttcctggca
      961 tttgtggtct tccttttagt gaccctagcg atactgacgg ccctccggct gtgcgcttat
     1021 tgttgtaaca ttgtcaacgt tagtctcgtg aagccctcgt tctacgttta ttcccgcgtg
     1081 aaaaatctaa acagctcccg tgtgcctgat cttcttgtct aaggatccgc aggcctctgc
     1141 tagcttgact gactgagata cagcgtacct tcagctcaca gacatgataa gatacattga
     1201 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     1261 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     1321 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     1381 aaacctctac aaatgtggta ttggcccatc tctatcggta tcgtagcata accccttggg
     1441 gcctctaaac gggtcttgag gggttttttg tgcccctcgg gccggattgc tatctaccgg
     1501 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
     1561 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
     1621 agaaatttat ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga
     1681 cattccttta atggtctttt ctggacacca ctaggggtca gaagtagttc atcaaacttt
     1741 cttccctccc taatctcatt ggttaccttg ggctatcgaa acttaattaa ccagtcaagt
     1801 cagctacttg gcgagatcga cttgtctggg tttcgactac gctcagaatt gcgtcagtca
     1861 agttcgatct ggtccttgct attgcacccg ttctccgatt acgagtttca tttaaatcat
     1921 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1981 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2041 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2101 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2161 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2221 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2281 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2341 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2401 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2461 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2521 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2581 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2641 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2701 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2761 acctatctca gcgatctgtc tatttcgttc atccatagtt gcatttaaat ttccgaactc
     2821 tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg caggctacct
     2881 ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc cttggctatt gcttggcagc
     2941 gcctatcgcc aggtattact ccaatcccga atatccgaga tcgggatcac ccgagagaag
     3001 ttcaacctac atcctcaatc ccgatctatc cgagatccga ggaatatcga aatcggggcg
     3061 cgcctggcct ccgcgccggg ttttggcgcc tcccgcgggc gcccccctcg tcacggcgag
     3121 cgctgccacg tcagacgaag ggcgcaggag cgtcctgatc cttccgcccg gacgctcagg
     3181 acagcggccc gctgctcata agactcggcc ttagaacccc agtatcagca gaaggacatt
     3241 ttaggacggg acttgggtga ctctagggca ctggttttct ttccagagag cggaacaggc
     3301 gaggaaaagt agtcccttct cggcgattct gcggagggat ctccgtgggg cggtgaacgc
     3361 cgatgattat ataaggacgc gccgggtgtg gcacagctag ttccgtcgca gccgggattt
     3421 gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag cgggctgctg
     3481 ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg gacggaagcg tgtggagaga
     3541 ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc ctgaactggg ggttgggggg
     3601 agcgcagcaa aatggcggct gttcccgagt cttgaatgga agacgcttgt gaggcgggct
     3661 gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc aagaacccaa ggtcttgagc
     3721 ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg gctgggcacc atctggggac
     3781 cctgacgtga agtttgtcac tgactggaga actcggtttg tcgtctgttg cgggggcggc
     3841 agttatggcg gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg ccctcgtcgt
     3901 gtcgtgacgt cacccgttct gttggcttat aatgcagggt ggggccacca gccggtaggt
     3961 gtgcggtagg cttttctccg tcgcaggacg cagggttcgg gcctagggta ggctctcctg
     4021 aatcgacagg cgccggacct ctggtgaggg gagggataag tgaggcgtca gtttctttgg
     4081 tcggttttat gtacctatct tcttaagtag ctgaagctcc ggttttgaac tatgcgctcg
     4141 gggttggcga gtgtgttttg tgaagttttt taggcacctt ttgaaatgta atcatttggg
     4201 tcaatatgta attttcagtg ttagacttgt aaattgtccg ctaaattctg gccgtttttg
     4261 gcttttttgt tagacaacat gggtaaaaag cctgaactca ccgcgacgtc tgtcgagaag
     4321 tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa
     4381 tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc
     4441 gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatccgc cgcgctcccg
     4501 attccggaag tgcttgacat tggggagttc agcgagagcc tgacctattg catctcccgc
     4561 cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag
     4621 ccggtcgcgg aggcaatgga tgccatcgct gccgccgatc ttagccagac gagcgggttc
     4681 ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg
     4741 attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc
     4801 gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac
     4861 ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg
     4921 gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc
     4981 ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat
     5041 ccggagcttg caggatcgcc ccggctccgg gcgtatatgc tccgcattgg tcttgaccaa
     5101 ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc
     5161 gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc
     5221 gccgccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc
     5281 agcactcgtc cgagggcaaa ggaataagct agtatgtaag cctagtctta gataataaaa
     5341 tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg tgtgtaacgc taggcgcgcc
     5401 tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg ttgcttggca
     5461 gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat attgtactca
     5521 agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt ctgatcggtc
     5581 aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat cagcaggagg
     5641 ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     5701 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     5761 gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     5821 tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     5881 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     5941 tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     6001 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     6061 aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     6121 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     6181 cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg aactacttac
     6241 tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg caggaccact
     6301 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     6361 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     6421 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     6481 aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc agaaaaaaat
     6541 gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg
     6601 gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta tccttaagat
     6661 cccgaatcgt ttaaactcga ctctggctct atcgaatctc cgtcgtttcg agcttacgcg
     6721 aacagccgtg gcgctcattt gctcgtcggg catcgaatct cgtca
//
