LOCUS       sequence-307379-        6513 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 6513)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6513)
  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-m4252944
FEATURES             Location/Qualifiers
     source          1..6513
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      225..361
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    513..584
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(589..635)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    653..701
                     /note="synthetic polyadenylation signal"
     enhancer        782..1085
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1087..1289
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1240..1260
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1589..1609
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1610..1654
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    1723..1844
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1734..1753)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1788..1807
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(2187..2775)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2267..2286)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3275..4492
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4329..4348
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4496..5524
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5551..5599
                     /note="synthetic polyadenylation signal"
     CDS             join(5865..5933,5934..6513,1..212)
                     /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(6083..6102)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
       61 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
      121 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
      181 agataggtgc ctcactgatt aagcattggt aaccgattct aggtgcattg gcgcagaaaa
      241 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
      301 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
      361 agatcccgaa tcgtttaaac tcgactctgg ctctatcgaa tctccgtcgt ttcgagctta
      421 cgcgaacagc cgtggcgctc atttgctcgt cgggcatcga atctcgtcag ctatcgtcag
      481 cttacctttt tggcagcgat cgcggctccc gacatcttgg accattagct ccacaggtat
      541 cttcttccct ctagtggtca taacagcagc ttcagctacc tctcaattca aaaaacccct
      601 caagacccgt ttagaggccc caaggggtta tgctatcaat cgttgcgtta cacacacaaa
      661 aaaccaacac acatccatct tcgatggata gcgattttat tatctaactg ctgatcgagt
      721 gtagccagat ctagtaatca attacggggt cattagttca tagcccatat atggagttcc
      781 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      841 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      901 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      961 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     1021 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     1081 ccatgctgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     1141 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     1201 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     1261 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc agatctttgt
     1321 cgatcctacc atccactcga cacacccgcc agcggccgcc accatgtact cattcgtttc
     1381 ggaagagaca ggtacgttaa tagttaatag cgtacttctt tttcttgctt tcgtggtatt
     1441 cttgctagtt acactagcca tccttactgc gcttcgattg tgtgcgtact gctgcaatat
     1501 tgttaacgtg agtcttgtaa aaccttcttt ttacgtttac tctcgtgtta aaaatctgaa
     1561 ttcttctaga gttcctgatc ttctggtcga aaacctttac tttcagtcga aagaaaccgc
     1621 ggcagctaag ttcgagaggc agcacatgga cagttaagga tccgcaggcc tctgctagct
     1681 tgactgactg agatacagcg taccttcagc tcacagacat gataagatac attgatgagt
     1741 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     1801 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     1861 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
     1921 tctacaaatg tggtattggc ccatctctat cggtatcgta gcataacccc ttggggcctc
     1981 tatatcgaaa cttaattaac cagtcaagtc agctacttgg cgagatcgac ttgtctgggt
     2041 ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt
     2101 tctccgatta cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac
     2161 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2221 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2281 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     2341 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     2401 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     2461 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     2521 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     2581 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     2641 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     2701 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     2761 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     2821 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     2881 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     2941 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3001 tccatagttg catttaaatt tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc
     3061 tttcgtccga tccatcttgc aggctacctc tcgaacgaac tatcgcaagt ctcttggccg
     3121 gccttgcgcc ttggctattg cttggcagcg cctatcgcca ggtattactc caatcccgaa
     3181 tatccgagat cgggatcacc cgagagaagt tcaacctaca tcctcaatcc cgatctatcc
     3241 gagatccgag gaatatcgaa atcggggcgc gcctggcctc cgcgccgggt tttggcgcct
     3301 cccgcgggcg cccccctcgt cacggcgagc gctgccacgt cagacgaagg gcgcaggagc
     3361 gtcctgatcc ttccgcccgg acgctcagga cagcggcccg ctgctcataa gactcggcct
     3421 tagaacccca gtatcagcag aaggacattt taggacggga cttgggtgac tctagggcac
     3481 tggttttctt tccagagagc ggaacaggcg aggaaaagta gtcccttctc ggcgattctg
     3541 cggagggatc tccgtggggc ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg
     3601 cacagctagt tccgtcgcag ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg
     3661 atcgtcactt ggtgagtagc gggctgctgg gctggccggg gctttcgtgg ccgccgggcc
     3721 gctcggtggg acggaagcgt gtggagagac cgccaagggc tgtagtctgg gtccgcgagc
     3781 aaggttgccc tgaactgggg gttgggggga gcgcagcaaa atggcggctg ttcccgagtc
     3841 ttgaatggaa gacgcttgtg aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg
     3901 gtgggcggca agaacccaag gtcttgagcc cttcgctaat gcgggaaagc tcttattcgg
     3961 gtgagatggg ctgggcacca tctggggacc ctgacgtgaa gtttgtcact gactggagaa
     4021 ctcggtttgt cgtctgttgc gggggcggca gttatggcgg tgccgttggg cagtgcaccc
     4081 gtacctttgg gagcgcgcgc cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata
     4141 atgcagggtg gggccacctg cagctcagcc ggtaggtgtg cggtaggctt ttctccgtcg
     4201 caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg
     4261 gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct
     4321 taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga
     4381 agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta
     4441 gacttgtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag acaacatggg
     4501 taaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag
     4561 cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt
     4621 aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg
     4681 ttatgtttat cggcactttg catccgccgc gctcccgatt ccggaagtgc ttgacattgg
     4741 ggagttcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca
     4801 agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg caatggatgc
     4861 catcgctgcc gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat
     4921 cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca
     4981 ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct
     5041 gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc
     5101 caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat
     5161 gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg
     5221 tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccccg
     5281 gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg
     5341 caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc
     5401 cgggactgtc gggcgtacac aaatcgcccg cagaagcgcc gccgtctgga ccgatggctg
     5461 tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga
     5521 ataagctagt atgtaagcct agtcttagat aataaaatcg ctatccatcg aagatggatg
     5581 tgtgttggtt ttttgtgtgt gtaacgctag gcgcgcctgg tgtaccgaga acgatcctct
     5641 cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt
     5701 gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg
     5761 atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt
     5821 ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt
     5881 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     5941 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg
     6001 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgytttccaa
     6061 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     6121 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag
     6181 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     6241 ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc
     6301 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     6361 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     6421 caacsttgcg caaactatta actggcgaac tacttactct agcttcccgg caacarttra
     6481 tagactggat ggaggcggat aaagttgcag gac
//
