LOCUS       sequence-307371-        6813 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 6813)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6813)
  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-m4252951
FEATURES             Location/Qualifiers
     source          1..6813
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      150..286
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    438..509
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(514..560)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    578..626
                     /note="synthetic polyadenylation signal"
     enhancer        707..1010
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1012..1214
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1165..1185
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1514..1534
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1535..1561
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1630..1751
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1641..1660)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1695..1714
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1866..1912
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1942..2078
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2123..2194
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2420..3008)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2500..2519)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3508..4717
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4554..4573
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4721..5749
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5776..5824
                     /note="synthetic polyadenylation signal"
     CDS             join(6090..6158,6159..6813,1..137)
                     /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(6308..6327)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
       61 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
      121 tgattaagca ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga
      181 cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac
      241 ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaagatc ccgaatcgtt
      301 taaactcgac tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg
      361 cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca
      421 gcgatcgcgg ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt
      481 ggtcataaca gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga
      541 ggccccaagg ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc
      601 catcttcgat ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt
      661 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      721 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      781 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      841 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      901 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      961 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg ctgatgcggt
     1021 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     1081 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     1141 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     1201 atataagcag agctggttta gtgaaccgtc agatcagatc tttgtcgatc ctaccatcca
     1261 ctcgacacac ccgccagcgg ccgccaccat gtacagcttt gtatctgaag agacaggcac
     1321 tctcatcgtt aattcagtgt tgctgttcct ggcatttgtg gtcttccttt tagtgaccct
     1381 agcgatactg acggccctcc ggctgtgcgc ttattgttgt aacattgtca acgttagtct
     1441 cgtgaagccc tcgttctacg tttattcccg cgtgaaaaat ctaaacagct cccgtgtgcc
     1501 tgatcttctt gtcgaaaacc tttactttca gtcgtacccc tatgacgtac cagattatgc
     1561 gtaaggatcc gcaggcctct gctagcttga ctgactgaga tacagcgtac cttcagctca
     1621 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     1681 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     1741 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     1801 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tattggccca tctctatcgg
     1861 tatcgtagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgtgcccctc
     1921 gggccggatt gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc
     1981 gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac
     2041 ttccatacgt gtcctcctta ccagaaattt atccttaagg tcgtcagcta tcctgcaggc
     2101 gatctctcga tttcgatcaa gacattcctt taatggtctt ttctggacac cactaggggt
     2161 cagaagtagt tcatcaaact ttcttccctc cctaatctca ttggttacct tgggctatcg
     2221 aaacttaatt aaccagtcaa gtcagctact tggcgagatc gacttgtctg ggtttcgact
     2281 acgctcagaa ttgcgtcagt caagttcgat ctggtccttg ctattgcacc cgttctccga
     2341 ttacgagttt catttaaatc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2401 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2461 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2521 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2581 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2641 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2701 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2761 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2821 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     2881 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2941 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     3001 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     3061 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     3121 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     3181 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     3241 ttgcatttaa atttccgaac tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc
     3301 cgatccatct tgcaggctac ctctcgaacg aactatcgca agtctcttgg ccggccttgc
     3361 gccttggcta ttgcttggca gcgcctatcg ccaggtatta ctccaatccc gaatatccga
     3421 gatcgggatc acccgagaga agttcaacct acatcctcaa tcccgatcta tccgagatcc
     3481 gaggaatatc gaaatcgggg cgcgcctggc ctccgcgccg ggttttggcg cctcccgcgg
     3541 gcgcccccct cgtcacggcg agcgctgcca cgtcagacga agggcgcagg agcgtcctga
     3601 tccttccgcc cggacgctca ggacagcggc ccgctgctca taagactcgg ccttagaacc
     3661 ccagtatcag cagaaggaca ttttaggacg ggacttgggt gactctaggg cactggtttt
     3721 ctttccagag agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg
     3781 atctccgtgg ggcggtgaac gccgatgatt atataaggac gcgccgggtg tggcacagct
     3841 agttccgtcg cagccgggat ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca
     3901 cttggtgagt agcgggctgc tgggctggcc ggggctttcg tggccgccgg gccgctcggt
     3961 gggacggaag cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg
     4021 ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg
     4081 gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg
     4141 gcaagaaccc aaggtcttga gcccttcgct aatgcgggaa agctcttatt cgggtgagat
     4201 gggctgggca ccatctgggg accctgacgt gaagtttgtc actgactgga gaactcggtt
     4261 tgtcgtctgt tgcgggggcg gcagttatgg cggtgccgtt gggcagtgca cccgtacctt
     4321 tgggagcgcg cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg
     4381 gtggggccac cagccggtag gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc
     4441 gggcctaggg taggctctcc tgaatcgaca ggcgccggac ctctggtgag gggagggata
     4501 agtgaggcgt cagtttcttt ggtcggtttt atgtacctat cttcttaagt agctgaagct
     4561 ccggttttga actatgcgct cggggttggc gagtgtgttt tgtgaagttt tttaggcacc
     4621 ttttgaaatg taatcatttg ggtcaatatg taattttcag tgttagactt gtaaattgtc
     4681 cgctaaattc tggccgtttt tggctttttt gttagacaac atgggtaaaa agcctgaact
     4741 caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat
     4801 gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata
     4861 tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca
     4921 ctttgcatcc gccgcgctcc cgattccgga agtgcttgac attggggagt tcagcgagag
     4981 cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac
     5041 cgaactgccc gctgttctgc agccggtcgc ggaggcaatg gatgccatcg ctgccgccga
     5101 tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac
     5161 atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat
     5221 ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga
     5281 ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac
     5341 ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca
     5401 atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac
     5461 gcgctacttc gagcggaggc atccggagct tgcaggatcg ccccggctcc gggcgtatat
     5521 gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc
     5581 agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg
     5641 tacacaaatc gcccgcagaa gcgccgccgt ctggaccgat ggctgtgtag aagtactcgc
     5701 cgatagtgga aaccgacgcc ccagcactcg tccgagggca aaggaataag ctagtatgta
     5761 agcctagtct tagataataa aatcgctatc catcgaagat ggatgtgtgt tggttttttg
     5821 tgtgtgtaac gctaggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga
     5881 cgatccatat cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc
     5941 caatcgtcag atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta
     6001 gatattgata gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc
     6061 aagagacagg atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta
     6121 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     6181 taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca
     6241 gcggtaagat ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta
     6301 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     6361 gccgcataca ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc
     6421 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     6481 ctgcggccaa cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc
     6541 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6601 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac
     6661 tattaactgg cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg
     6721 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6781 ataaatctgg agccggtgag cgtgggtctc gcg
//
