LOCUS       sequence-300622-        4461 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 4461)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4461)
  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-m4133425
FEATURES             Location/Qualifiers
     source          1..4461
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      55..101
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    106..177
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(329..465)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(478..1269,1270..1338))
                     /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     1101..1120
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2084..2672
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2573..2592
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    2898..2969
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(3014..3150)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(3180..3226)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    3341..3462
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3378..3397)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3432..3451
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(3515..3541)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(3542..3562)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(3862..4064)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3891..3911)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4066..4369
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    join(4450..4461,1..37)
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 atccatcgaa gatggatgtg tgttggtttt ttgtgtgtgt aacgcaacga ttgatagcat
       61 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gaattgagag gtagctgaag
      121 ctgctgttat gaccactaga gggaagaaga tacctgtgga gctaatggtc caagatgtcg
      181 ggagccgcga tcgctgccaa aaaggtaagc tgacgatagc tgacgagatt cgatgcccga
      241 cgagcaaatg agcgccacgg ctgttcgcgt aagctcgaaa cgacggagat tcgatagagc
      301 cagagtcgag tttaaacgat tcgggatctt aaggataaat ttctggtaag gaggacacgt
      361 atggaagtgg gcaagttggg gaagccgtat ccgttgctga atctggcata tgtgggagta
      421 taagacgcgc agcgtcgcat caggcatttt tttctgcgcc aatgcaccta gaatcggtta
      481 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
      541 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
      601 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
      661 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
      721 tatcaactgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tacgcaaggt
      781 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
      841 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
      901 tagctccttc ggtcctccaa tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
      961 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     1021 gactggtgaa tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     1081 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     1141 cattggaaag cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     1201 ttcgatgtaa cccactcgcg cacccaactg atcttcagca tcttttactt tcaccagcgt
     1261 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     1321 gaaatgttga atactcatgc gaaagcctcc tgctgatcct gtctcttgat agatgtttgc
     1381 aaaagctagg actcgattat acgttgaccg atcagactat caatatctag gtttaaacag
     1441 atcggtacgc tgccgtgacc aggcttgagt acaatatctg acgattggac ttcctgggtc
     1501 ccaagctgga ttccgactgg ctgactgcca agcaacgata tggatcgtcg agactcgcac
     1561 tgagaggatc gttctcggta caccaggcgc gccccgattt cgatattcct cggatctcgg
     1621 atagatcggg attgaggatg taggttgaac ttctctcggg tgatcccgat ctcggatatt
     1681 cgggattgga gtaatacctg gcgataggcg ctgccaagca atagccaagg cgcaaggccg
     1741 gccaagagac ttgcgatagt tcgttcgaga ggtagcctgc aagatggatc ggacgaaagg
     1801 tttgaagatt ttccgacgag ggccttggag agttcggaaa tttaaatgca actatggatg
     1861 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     1921 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     1981 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     2041 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     2101 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     2161 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     2221 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     2281 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     2341 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     2401 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     2461 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     2521 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     2581 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     2641 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     2701 tcctggcctt ttgctggcct tttgctcaca tgatttaaat gaaactcgta atcggagaac
     2761 gggtgcaata gcaaggacca gatcgaactt gactgacgca attctgagcg tagtcgaaac
     2821 ccagacaagt cgatctcgcc aagtagctga cttgactggt taattaagtt tcgatagccc
     2881 aaggtaacca atgagattag ggagggaaga aagtttgatg aactacttct gacccctagt
     2941 ggtgtccaga aaagaccatt aaaggaatgt cttgatcgaa atcgagagat cgcctgcagg
     3001 atagctgacg accttaagga taaatttctg gtaaggagga cacgtatgga agtgggcaag
     3061 ttggggaagc cgtatccgtt gctgaatctg gcatatgtgg gagtataaga cgcgcagcgt
     3121 cgcatcaggc atttttttct gcgccaatgc cggtagatag caatccggcc cgaggggcac
     3181 aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctacgat accgatagag
     3241 atgggccaat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     3301 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     3361 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3421 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtgagctgaa
     3481 ggtacgctgt atctcagtca gtcaagctag cttacgcata atctggtacg tcataggggt
     3541 acgactgaaa gtaaaggttt tcgaccagaa gatcaggaac tctagaagaa ttcagatttt
     3601 taacacgaga gtaaacgtaa aaagaaggtt ttacaagact cacgttaaca atattgcagc
     3661 agtacgcaca caatcgaagc gcagtaagga tggctagtgt aactagcaag aataccacga
     3721 aagcaagaaa aagaagtacg ctattaacta ttaacgtacc tgtctcttcc gaaacgaatg
     3781 agtacatggt ggcggccgct ggcgggtgtg tcgagtggat ggtaggatcg acaaagatct
     3841 gatctgacgg ttcactaaac cagctctgct tatatagacc tcccaccgta cacgcctacc
     3901 gcccatttgc gtcaatgggg cggagttgtt acgacatttt ggaaagtccc gttgattttg
     3961 gtgccaaaac aaactcccat tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc
     4021 aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc atcagcatgg taatagcgat
     4081 gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata
     4141 atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata
     4201 cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg
     4261 gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg
     4321 gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat
     4381 atatgggcta tgaactaatg accccgtaat tgattactag atctggctac actcgatcag
     4441 cagttagata ataaaatcgc t
//
