LOCUS       sequence-300702-        6828 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 6828)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6828)
  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-m4252873
FEATURES             Location/Qualifiers
     source          1..6828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    59..180
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(70..89)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     124..143
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      295..341
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      371..507
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    552..623
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(849..1437)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(929..948)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1937..3146
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     2983..3002
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3150..4178
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4205..4253
                     /note="synthetic polyadenylation signal"
     CDS             join(4519..4587,4588..5379)
                     /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(4737..4756)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5392..5528
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5680..5751
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(5756..5802)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    5820..5868
                     /note="synthetic polyadenylation signal"
     enhancer        5949..6252
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6254..6456
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6407..6427
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             6756..6776
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             6777..6818
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
ORIGIN
        1 caggcctctg ctagcttgac tgactgagat acagcgtacc ttcagctcac agacatgata
       61 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
      121 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
      181 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
      241 taaagcaagt aaaacctcta caaatgtggt attggcccat ctctatcggt atcgtagcat
      301 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg ggccggattg
      361 ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc
      421 ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact tccatacgtg
      481 tcctccttac cagaaattta tccttaaggt cgtcagctat cctgcaggcg atctctcgat
      541 ttcgatcaag acattccttt aatggtcttt tctggacacc actaggggtc agaagtagtt
      601 catcaaactt tcttccctcc ctaatctcat tggttacctt gggctatcga aacttaatta
      661 accagtcaag tcagctactt ggcgagatcg acttgtctgg gtttcgacta cgctcagaat
      721 tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc gttctccgat tacgagtttc
      781 atttaaatca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
      841 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
      901 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
      961 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     1021 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     1081 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     1141 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     1201 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     1261 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     1321 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     1381 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     1441 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     1501 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     1561 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     1621 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcatttaaa
     1681 tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc gatccatctt
     1741 gcaggctacc tctcgaacga actatcgcaa gtctcttggc cggccttgcg ccttggctat
     1801 tgcttggcag cgcctatcgc caggtattac tccaatcccg aatatccgag atcgggatca
     1861 cccgagagaa gttcaaccta catcctcaat cccgatctat ccgagatccg aggaatatcg
     1921 aaatcggggc gcgcctggcc tccgcgccgg gttttggcgc ctcccgcggg cgcccccctc
     1981 gtcacggcga gcgctgccac gtcagacgaa gggcgcagga gcgtcctgat ccttccgccc
     2041 ggacgctcag gacagcggcc cgctgctcat aagactcggc cttagaaccc cagtatcagc
     2101 agaaggacat tttaggacgg gacttgggtg actctagggc actggttttc tttccagaga
     2161 gcggaacagg cgaggaaaag tagtcccttc tcggcgattc tgcggaggga tctccgtggg
     2221 gcggtgaacg ccgatgatta tataaggacg cgccgggtgt ggcacagcta gttccgtcgc
     2281 agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac ttggtgagta
     2341 gcgggctgct gggctggccg gggctttcgt ggccgccggg ccgctcggtg ggacggaagc
     2401 gtgtggagag accgccaagg gctgtagtct gggtccgcga gcaaggttgc cctgaactgg
     2461 gggttggggg gagcgcagca aaatggcggc tgttcccgag tcttgaatgg aagacgcttg
     2521 tgaggcgggc tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg caagaaccca
     2581 aggtcttgag cccttcgcta atgcgggaaa gctcttattc gggtgagatg ggctgggcac
     2641 catctgggga ccctgacgtg aagtttgtca ctgactggag aactcggttt gtcgtctgtt
     2701 gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc
     2761 gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta taatgcaggg tggggccacc
     2821 agccggtagg tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg ggcctagggt
     2881 aggctctcct gaatcgacag gcgccggacc tctggtgagg ggagggataa gtgaggcgtc
     2941 agtttctttg gtcggtttta tgtacctatc ttcttaagta gctgaagctc cggttttgaa
     3001 ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt
     3061 aatcatttgg gtcaatatgt aattttcagt gttagacttg taaattgtcc gctaaattct
     3121 ggccgttttt ggcttttttg ttagacaaca tgggtaaaaa gcctgaactc accgcgacgt
     3181 ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctcgg
     3241 agggcgaaga atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg
     3301 taaatagctg cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatccg
     3361 ccgcgctccc gattccggaa gtgcttgaca ttggggagtt cagcgagagc ctgacctatt
     3421 gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg
     3481 ctgttctgca gccggtcgcg gaggcaatgg atgccatcgc tgccgccgat cttagccaga
     3541 cgagcgggtt cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt
     3601 tcatatgcgc gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg
     3661 tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg
     3721 aagtccggca cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc
     3781 gcataacagc ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg
     3841 ccaacatctt cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg
     3901 agcggaggca tccggagctt gcaggatcgc cccggctccg ggcgtatatg ctccgcattg
     3961 gtcttgacca actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc
     4021 agggtcgatg cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg
     4081 cccgcagaag cgccgccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa
     4141 accgacgccc cagcactcgt ccgagggcaa aggaataagc tagtatgtaa gcctagtctt
     4201 agataataaa atcgctatcc atcgaagatg gatgtgtgtt ggttttttgt gtgtgtaacg
     4261 ctaggcgcgc ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac gatccatatc
     4321 gttgcttggc agtcagccag tcggaatcca gcttgggacc caggaagtcc aatcgtcaga
     4381 tattgtactc aagcctggtc acggcagcgt accgatctgt ttaaacctag atattgatag
     4441 tctgatcggt caacgtataa tcgagtccta gcttttgcaa acatctatca agagacagga
     4501 tcagcaggag gctttcgcat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4561 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4621 gaagatcagt tgggtgcgcg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4681 cttgagagtt ttcgccccga agaacgcttt ccaatgatga gcacttttaa agttctgcta
     4741 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     4801 tattctcaga atgacttggt tgagtattca ccagtcacag aaaagcatct tacggatggc
     4861 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     4921 ttacttctga caacgattgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     4981 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5041 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacct tgcgtaaact attaactggc
     5101 gaactactta ctctagcttc ccggcaacag ttgatagact ggatggaggc ggataaagtt
     5161 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5221 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5281 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5341 atcgctgaga taggtgcctc actgattaag cattggtaac cgattctagg tgcattggcg
     5401 cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg ccagattcag
     5461 caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta ccagaaattt
     5521 atccttaaga tcccgaatcg tttaaactcg actctggctc tatcgaatct ccgtcgtttc
     5581 gagcttacgc gaacagccgt ggcgctcatt tgctcgtcgg gcatcgaatc tcgtcagcta
     5641 tcgtcagctt acctttttgg cagcgatcgc ggctcccgac atcttggacc attagctcca
     5701 caggtatctt cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa
     5761 aacccctcaa gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac
     5821 acacaaaaaa ccaacacaca tccatcttcg atggatagcg attttattat ctaactgctg
     5881 atcgagtgta gccagatcta gtaatcaatt acggggtcat tagttcatag cccatatatg
     5941 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     6001 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     6061 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     6121 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     6181 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     6241 gctattacca tgctgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     6301 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     6361 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     6421 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatcaga
     6481 tctttgtcga tcctaccatc cactcgacac acccgccagc ggccgccacc atgtactcat
     6541 tcgtttcgga agagacaggt acgttaatag ttaatagcgt acttcttttt cttgctttcg
     6601 tggtattctt gctagttaca ctagccatcc ttactgcgct tcgattgtgt gcgtactgct
     6661 gcaatattgt taacgtgagt cttgtaaaac cttcttttta cgtttactct cgtgttaaaa
     6721 atctgaattc ttctagagtt cctgatcttc tggtcgaaaa cctttacttt cagtcgggga
     6781 aacctatacc caatccactt ctcggcttag attctaccta aggatccg
//
