LOCUS       sequence-300689-        7176 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 7176)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7176)
  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-m4252919
FEATURES             Location/Qualifiers
     source          1..7176
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    211..259
                     /note="synthetic polyadenylation signal"
     terminator      277..323
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    328..399
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(551..687)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             complement(join(700..1491,1492..1560))
                     /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     1323..1342
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1826..1874
                     /note="synthetic polyadenylation signal"
     CDS             complement(1901..2929)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     promoter        complement(2933..4142)
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     complement(3077..3096)
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     rep_origin      4642..5230
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5131..5150
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5456..5527
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5572..5708)
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     terminator      complement(5738..5784)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5899..6020
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5936..5955)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5990..6009
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(6113..6130)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(6479..6499)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(6799..7001)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(6828..6848)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        join(7003..7176,1..130)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat
       61 tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag
      121 ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta
      181 gatctggcta cactcgatca gcagttagat aataaaatcg ctatccatcg aagatggatg
      241 tgtgttggtt ttttgtgtgt gtaacgcaac gattgatagc ataacccctt ggggcctcta
      301 aacgggtctt gaggggtttt ttgaattgag aggtagctga agctgctgtt atgaccacta
      361 gagggaagaa gatacctgtg gagctaatgg tccaagatgt cgggagccgc gatcgctgcc
      421 aaaaaggtaa gctgacgata gctgacgaga ttcgatgccc gacgagcaaa tgagcgccac
      481 ggctgttcgc gtaagctcga aacgacggag attcgataga gccagagtcg agtttaaacg
      541 attcgggatc ttaaggataa atttctggta aggaggacac gtatggaagt gggcaagttg
      601 gggaagccgt atccgttgct gaatctggca tatgtgggag tataagacgc gcagcgtcgc
      661 atcaggcatt tttttctgcg ccaatgcacc tagaatcggt taccaatgct taatcagtga
      721 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
      781 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
      841 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
      901 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctatcaact gttgccggga
      961 agctagagta agtagttcgc cagttaatag tttacgcaag gttgttgcca ttgctacagg
     1021 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     1081 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     1141 aatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     1201 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg aatactcaac
     1261 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     1321 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa agcgttcttc
     1381 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     1441 cgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     1501 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     1561 gcgaaagcct cctgctgatc ctgtctcttg atagatgttt gcaaaagcta ggactcgatt
     1621 atacgttgac cgatcagact atcaatatct aggtttaaac agatcggtac gctgccgtga
     1681 ccaggcttga gtacaatatc tgacgattgg acttcctggg tcccaagctg gattccgact
     1741 ggctgactgc caagcaacga tatggatcgt cgagactcgc actgagagga tcgttctcgg
     1801 tacaccaggc gcgcctagcg ttacacacac aaaaaaccaa cacacatcca tcttcgatgg
     1861 atagcgattt tattatctaa gactaggctt acatactagc ttattccttt gccctcggac
     1921 gagtgctggg gcgtcggttt ccactatcgg cgagtacttc tacacagcca tcggtccaga
     1981 cggcggcgct tctgcgggcg atttgtgtac gcccgacagt cccggctccg gatcggacga
     2041 ttgcgtcgca tcgaccctgc gcccaagctg catcatcgaa attgccgtca accaagctct
     2101 gatagagttg gtcaagacca atgcggagca tatacgcccg gagccggggc gatcctgcaa
     2161 gctccggatg cctccgctcg aagtagcgcg tctgctgctc catacaagcc aaccacggcc
     2221 tccagaagaa gatgttggcg acctcgtatt gggaatcccc gaacatcgcc tcgctccagt
     2281 caatgaccgc tgttatgcgg ccattgtccg tcaggacatt gttggagccg aaatccgcgt
     2341 gcacgaggtg ccggacttcg gggcagtcct cggcccaaag catcagctca tcgagagcct
     2401 gcgcgacgga cgcactgacg gtgtcgtcca tcacagtttg ccagtgatac acatggggat
     2461 cagcaatcgc gcatatgaaa tcacgccatg tagtgtattg accgattcct tgcggtccga
     2521 atgggccgaa cccgctcgtc tggctaagat cggcggcagc gatggcatcc attgcctccg
     2581 cgaccggctg cagaacagcg ggcagttcgg tttcaggcag gtcttgcaac gtgacaccct
     2641 gtgcacggcg ggagatgcaa taggtcaggc tctcgctgaa ctccccaatg tcaagcactt
     2701 ccggaatcgg gagcgcggcg gatgcaaagt gccgataaac ataacgatct ttgtagaaac
     2761 catcggcgca gctatttacc cgcaggacat atccacgccc tcctacatcg aagctgaaag
     2821 cacgagattc ttcgccctcc gagagctgca tcaggtcgga gacgctgtcg aacttttcga
     2881 tcagaaactt ctcgacagac gtcgcggtga gttcaggctt tttacccatg ttgtctaaca
     2941 aaaaagccaa aaacggccag aatttagcgg acaatttaca agtctaacac tgaaaattac
     3001 atattgaccc aaatgattac atttcaaaag gtgcctaaaa aacttcacaa aacacactcg
     3061 ccaaccccga gcgcatagtt caaaaccgga gcttcagcta cttaagaaga taggtacata
     3121 aaaccgacca aagaaactga cgcctcactt atccctcccc tcaccagagg tccggcgcct
     3181 gtcgattcag gagagcctac cctaggcccg aaccctgcgt cctgcgacgg agaaaagcct
     3241 accgcacacc taccggctgg tggccccacc ctgcattata agccaacaga acgggtgacg
     3301 tcacgacacg acgagggcgc gcgctcccaa aggtacgggt gcactgccca acggcaccgc
     3361 cataactgcc gcccccgcaa cagacgacaa accgagttct ccagtcagtg acaaacttca
     3421 cgtcagggtc cccagatggt gcccagccca tctcacccga ataagagctt tcccgcatta
     3481 gcgaagggct caagaccttg ggttcttgcc gcccaccatg ccccccacct tgtttcaacg
     3541 acctcacagc ccgcctcaca agcgtcttcc attcaagact cgggaacagc cgccattttg
     3601 ctgcgctccc cccaaccccc agttcagggc aaccttgctc gcggacccag actacagccc
     3661 ttggcggtct ctccacacgc ttccgtccca ccgagcggcc cggcggccac gaaagccccg
     3721 gccagcccag cagcccgcta ctcaccaagt gacgatcaca gcgatccaca aacaagaacc
     3781 gcgacccaaa tcccggctgc gacggaacta gctgtgccac acccggcgcg tccttatata
     3841 atcatcggcg ttcaccgccc cacggagatc cctccgcaga atcgccgaga agggactact
     3901 tttcctcgcc tgttccgctc tctggaaaga aaaccagtgc cctagagtca cccaagtccc
     3961 gtcctaaaat gtccttctgc tgatactggg gttctaaggc cgagtcttat gagcagcggg
     4021 ccgctgtcct gagcgtccgg gcggaaggat caggacgctc ctgcgccctt cgtctgacgt
     4081 ggcagcgctc gccgtgacga ggggggcgcc cgcgggaggc gccaaaaccc ggcgcggagg
     4141 ccaggcgcgc cccgatttcg atattcctcg gatctcggat agatcgggat tgaggatgta
     4201 ggttgaactt ctctcgggtg atcccgatct cggatattcg ggattggagt aatacctggc
     4261 gataggcgct gccaagcaat agccaaggcg caaggccggc caagagactt gcgatagttc
     4321 gttcgagagg tagcctgcaa gatggatcgg acgaaaggtt tgaagatttt ccgacgaggg
     4381 ccttggagag ttcggaaatt taaatgcaac tatggatgaa cgaaatagac agatcgctga
     4441 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     4501 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     4561 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     4621 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4681 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4741 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     4801 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4861 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4921 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4981 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5041 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5101 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     5161 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5221 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5281 tgctcacatg atttaaatga aactcgtaat cggagaacgg gtgcaatagc aaggaccaga
     5341 tcgaacttga ctgacgcaat tctgagcgta gtcgaaaccc agacaagtcg atctcgccaa
     5401 gtagctgact tgactggtta attaagtttc gatagcccaa ggtaaccaat gagattaggg
     5461 agggaagaaa gtttgatgaa ctacttctga cccctagtgg tgtccagaaa agaccattaa
     5521 aggaatgtct tgatcgaaat cgagagatcg cctgcaggat agctgacgac cttaaggata
     5581 aatttctggt aaggaggaca cgtatggaag tgggcaagtt ggggaagccg tatccgttgc
     5641 tgaatctggc atatgtggga gtataagacg cgcagcgtcg catcaggcat ttttttctgc
     5701 gccaatgccg gtagatagca atccggcccg aggggcacaa aaaacccctc aagacccgtt
     5761 tagaggcccc aaggggttat gctacgatac cgatagagat gggccaatac cacatttgta
     5821 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     5881 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5941 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     6001 aaactcatca atgtatctta tcatgtctgt gagctgaagg tacgctgtat ctcagtcagt
     6061 caagctagca gaggcctgcg gatccttagc cgcgtggcac gagcccgctg ctatggtgat
     6121 ggtgatggtg catgtgcccc gagcctgaac ctgccaggtt agcgtccaag aactctttta
     6181 gctggccctt agacagggct cccacttttg tcgcggcaac ttccccattc ttaaagagca
     6241 ggagggtagg tattcccctg atgccgtatt taggtgccgt ccctgggttc tgatctatat
     6301 taagcttagc gacagttaat tttccttgat actcgtcggc gatctcgtcc agaatgggtg
     6361 cgatcatctt gcacgggcca caccattcag cccagaaatc taccagtatc gcaccatcgg
     6421 ccttcaacac gtctgtatca aaggagtcat cggttagatg aatgattttg tcactcatcg
     6481 actgaaagta aaggttttcg accagaagat caggaactct agaagaattc agatttttaa
     6541 cacgagagta aacgtaaaaa gaaggtttta caagactcac gttaacaata ttgcagcagt
     6601 acgcacacaa tcgaagcgca gtaaggatgg ctagtgtaac tagcaagaat accacgaaag
     6661 caagaaaaag aagtacgcta ttaactatta acgtacctgt ctcttccgaa acgaatgagt
     6721 acatggtggc ggccgctggc gggtgtgtcg agtggatggt aggatcgaca aagatctgat
     6781 ctgacggttc actaaaccag ctctgcttat atagacctcc caccgtacac gcctaccgcc
     6841 catttgcgtc aatggggcgg agttgttacg acattttgga aagtcccgtt gattttggtg
     6901 ccaaaacaaa ctcccattga cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa
     6961 ccgctatcca cgcccattga tgtactgcca aaaccgcatc agcatggtaa tagcgatgac
     7021 taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg
     7081 ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac
     7141 ttgatgtact gccaagtggg cagtttaccg taaata
//
