LOCUS       sequence-313659-        6386 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 6386)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6386)
  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-m4252821
FEATURES             Location/Qualifiers
     source          1..6386
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             808..828
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             829..873
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    926..1047
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(937..956)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     991..1010
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1162..1208
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1238..1374
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1419..1490
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1716..2304)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1796..1815)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2804..4013
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3850..3869
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4017..4616
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4017..4036)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4513..4533
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4643..4691
                     /note="synthetic polyadenylation signal"
     CDS             join(4957..5025,5026..5817)
                     /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(5175..5194)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5830..5966
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6118..6189
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6194..6240)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6258..6306
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
      541 gatcctacca tccactcgac acacccgcca gcggccgcca ccatgtacag ctttgtatct
      601 gaagagacag gcactctcat cgttaattca gtgttgctgt tcctggcatt tgtggtcttc
      661 cttttagtga ccctagcgat actgacggcc ctccggctgt gcgcttattg ttgtaacatt
      721 gtcaacgtta gtctcgtgaa gccctcgttc tacgtttatt cccgcgtgaa aaatctaaac
      781 agctcccgtg tgcctgatct tcttgtcgaa aacctttact ttcagtcgaa agaaaccgcg
      841 gcagctaagt tcgagaggca gcacatggac agttaagcta gcttgactga ctgagataca
      901 gcgtaccttc agctcacaga catgataaga tacattgatg agtttggaca aaccacaact
      961 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     1021 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     1081 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatt
     1141 ggcccatctc tatcggtatc gtagcataac cccttggggc ctctaaacgg gtcttgaggg
     1201 gttttttgtg cccctcgggc cggattgcta tctaccggca ttggcgcaga aaaaaatgcc
     1261 tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct
     1321 tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt
     1381 cagctatcct gcaggcgatc tctcgatttc gatcaagaca ttcctttaat ggtcttttct
     1441 ggacaccact aggggtcaga agtagttcat caaactttct tccctcccta atctcattgg
     1501 ttaccttggg ctatcgaaac ttaattaacc agtcaagtca gctacttggc gagatcgact
     1561 tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg tccttgctat
     1621 tgcacccgtt ctccgattac gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag
     1681 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     1741 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     1801 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     1861 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     1921 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     1981 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2041 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2101 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     2161 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     2221 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     2281 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     2341 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     2401 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     2461 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     2521 tttcgttcat ccatagttgc atttaaattt ccgaactctc caaggccctc gtcggaaaat
     2581 cttcaaacct ttcgtccgat ccatcttgca ggctacctct cgaacgaact atcgcaagtc
     2641 tcttggccgg ccttgcgcct tggctattgc ttggcagcgc ctatcgccag gtattactcc
     2701 aatcccgaat atccgagatc gggatcaccc gagagaagtt caacctacat cctcaatccc
     2761 gatctatccg agatccgagg aatatcgaaa tcggggcgcg cctggcctcc gcgccgggtt
     2821 ttggcgcctc ccgcgggcgc ccccctcgtc acggcgagcg ctgccacgtc agacgaaggg
     2881 cgcaggagcg tcctgatcct tccgcccgga cgctcaggac agcggcccgc tgctcataag
     2941 actcggcctt agaaccccag tatcagcaga aggacatttt aggacgggac ttgggtgact
     3001 ctagggcact ggttttcttt ccagagagcg gaacaggcga ggaaaagtag tcccttctcg
     3061 gcgattctgc ggagggatct ccgtggggcg gtgaacgccg atgattatat aaggacgcgc
     3121 cgggtgtggc acagctagtt ccgtcgcagc cgggatttgg gtcgcggttc ttgtttgtgg
     3181 atcgctgtga tcgtcacttg gtgagtagcg ggctgctggg ctggccgggg ctttcgtggc
     3241 cgccgggccg ctcggtggga cggaagcgtg tggagagacc gccaagggct gtagtctggg
     3301 tccgcgagca aggttgccct gaactggggg ttggggggag cgcagcaaaa tggcggctgt
     3361 tcccgagtct tgaatggaag acgcttgtga ggcgggctgt gaggtcgttg aaacaaggtg
     3421 gggggcatgg tgggcggcaa gaacccaagg tcttgagccc ttcgctaatg cgggaaagct
     3481 cttattcggg tgagatgggc tgggcaccat ctggggaccc tgacgtgaag tttgtcactg
     3541 actggagaac tcggtttgtc gtctgttgcg ggggcggcag ttatggcggt gccgttgggc
     3601 agtgcacccg tacctttggg agcgcgcgcc ctcgtcgtgt cgtgacgtca cccgttctgt
     3661 tggcttataa tgcagggtgg ggccaccagc cggtaggtgt gcggtaggct tttctccgtc
     3721 gcaggacgca gggttcgggc ctagggtagg ctctcctgaa tcgacaggcg ccggacctct
     3781 ggtgagggga gggataagtg aggcgtcagt ttctttggtc ggttttatgt acctatcttc
     3841 ttaagtagct gaagctccgg ttttgaacta tgcgctcggg gttggcgagt gtgttttgtg
     3901 aagtttttta ggcacctttt gaaatgtaat catttgggtc aatatgtaat tttcagtgtt
     3961 agacttgtaa attgtccgct aaattctggc cgtttttggc ttttttgtta gacaacatga
     4021 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca
     4081 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc
     4141 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     4201 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     4261 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     4321 ggctggccgc gcagcaacag atggagggcc tcctggcgcc gcaccggccc aaggagcccg
     4381 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
     4441 ccgtcgtgct ccccggagtg gaggccgccg agcgcgccgg ggtgcccgcc ttcctggaga
     4501 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     4561 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctaggcta
     4621 gtatgtaagc ctagtcttag ataataaaat cgctatccat cgaagatgga tgtgtgttgg
     4681 ttttttgtgt gtgtaacgct aggcgcgcct ggtgtaccga gaacgatcct ctcagtgcga
     4741 gtctcgacga tccatatcgt tgcttggcag tcagccagtc ggaatccagc ttgggaccca
     4801 ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac cgatctgttt
     4861 aaacctagat attgatagtc tgatcggtca acgtataatc gagtcctagc ttttgcaaac
     4921 atctatcaag agacaggatc agcaggaggc tttcgcatga gtattcaaca tttccgtgtc
     4981 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5041 gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag tgggttacat cgaactggat
     5101 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgctttcc aatgatgagc
     5161 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     5221 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtattcacc agtcacagaa
     5281 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5341 gataacactg cggccaactt acttctgaca acgattggag gaccgaagga gctaaccgct
     5401 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5461 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaaccttg
     5521 cgtaaactat taactggcga actacttact ctagcttccc ggcaacagtt gatagactgg
     5581 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5641 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     5701 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     5761 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaaccg
     5821 attctaggtg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc
     5881 cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt
     5941 cctccttacc agaaatttat ccttaagatc ccgaatcgtt taaactcgac tctggctcta
     6001 tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg cgctcatttg ctcgtcgggc
     6061 atcgaatctc gtcagctatc gtcagcttac ctttttggca gcgatcgcgg ctcccgacat
     6121 cttggaccat tagctccaca ggtatcttct tccctctagt ggtcataaca gcagcttcag
     6181 ctacctctca attcaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta
     6241 tcaatcgttg cgttacacac acaaaaaacc aacacacatc catcttcgat ggatagcgat
     6301 tttattatct aactgctgat cgagtgtagc cagatctagt aatcaattac ggggtcatta
     6361 gttcatagcc catatatgga gttccg
//
