LOCUS       sequence-301681-        6855 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 6855)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6855)
  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-m4252875
FEATURES             Location/Qualifiers
     source          1..6855
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    142..213
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(218..264)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    282..330
                     /note="synthetic polyadenylation signal"
     enhancer        411..714
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        716..918
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     869..889
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1218..1238
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1242..1307
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1376..1497
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1387..1406)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1441..1460
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1612..1658
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1688..1824
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1869..1940
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2166..2754)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2246..2265)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3254..4463
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4300..4319
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4467..5495
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5522..5570
                     /note="synthetic polyadenylation signal"
     CDS             join(5836..5904,5905..6696)
                     /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(6054..6073)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6709..6845
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 cgtttaaact cgactctggc tctatcgaat ctccgtcgtt tcgagcttac gcgaacagcc
       61 gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc ttaccttttt
      121 ggcagcgatc gcggctcccg acatcttgga ccattagctc cacaggtatc ttcttccctc
      181 tagtggtcat aacagcagct tcagctacct ctcaattcaa aaaacccctc aagacccgtt
      241 tagaggcccc aaggggttat gctatcaatc gttgcgttac acacacaaaa aaccaacaca
      301 catccatctt cgatggatag cgattttatt atctaactgc tgatcgagtg tagccagatc
      361 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      421 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      481 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      541 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      601 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      661 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catgctgatg
      721 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      781 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      841 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
      901 gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc gatcctacca
      961 tccactcgac acacccgcca gcggccgcca ccatgtactc attcgtttcg gaagagacag
     1021 gtacgttaat agttaatagc gtacttcttt ttcttgcttt cgtggtattc ttgctagtta
     1081 cactagccat ccttactgcg cttcgattgt gtgcgtactg ctgcaatatt gttaacgtga
     1141 gtcttgtaaa accttctttt tacgtttact ctcgtgttaa aaatctgaat tcttctagag
     1201 ttcctgatct tctggtcgaa aacctttact ttcagtcgat ggattataag gaccacgatg
     1261 gagactacaa agatcatgac attgattaca aggacgacga tgacaaataa ggatccgcag
     1321 gcctctgcta gcttgactga ctgagataca gcgtaccttc agctcacaga catgataaga
     1381 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     1441 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     1501 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     1561 agcaagtaaa acctctacaa atgtggtatt ggcccatctc tatcggtatc gtagcataac
     1621 cccttggggc ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc cggattgcta
     1681 tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca
     1741 catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc
     1801 tccttaccag aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc
     1861 gatcaagaca ttcctttaat ggtcttttct ggacaccact aggggtcaga agtagttcat
     1921 caaactttct tccctcccta atctcattgg ttaccttggg ctatcgaaac ttaattaacc
     1981 agtcaagtca gctacttggc gagatcgact tgtctgggtt tcgactacgc tcagaattgc
     2041 gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt ctccgattac gagtttcatt
     2101 taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2161 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2221 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2281 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2341 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2401 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2461 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2521 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2581 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2641 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2701 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2761 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     2821 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2881 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     2941 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc atttaaattt
     3001 ccgaactctc caaggccctc gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca
     3061 ggctacctct cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct tggctattgc
     3121 ttggcagcgc ctatcgccag gtattactcc aatcccgaat atccgagatc gggatcaccc
     3181 gagagaagtt caacctacat cctcaatccc gatctatccg agatccgagg aatatcgaaa
     3241 tcggggcgcg cctggcctcc gcgccgggtt ttggcgcctc ccgcgggcgc ccccctcgtc
     3301 acggcgagcg ctgccacgtc agacgaaggg cgcaggagcg tcctgatcct tccgcccgga
     3361 cgctcaggac agcggcccgc tgctcataag actcggcctt agaaccccag tatcagcaga
     3421 aggacatttt aggacgggac ttgggtgact ctagggcact ggttttcttt ccagagagcg
     3481 gaacaggcga ggaaaagtag tcccttctcg gcgattctgc ggagggatct ccgtggggcg
     3541 gtgaacgccg atgattatat aaggacgcgc cgggtgtggc acagctagtt ccgtcgcagc
     3601 cgggatttgg gtcgcggttc ttgtttgtgg atcgctgtga tcgtcacttg gtgagtagcg
     3661 ggctgctggg ctggccgggg ctttcgtggc cgccgggccg ctcggtggga cggaagcgtg
     3721 tggagagacc gccaagggct gtagtctggg tccgcgagca aggttgccct gaactggggg
     3781 ttggggggag cgcagcaaaa tggcggctgt tcccgagtct tgaatggaag acgcttgtga
     3841 ggcgggctgt gaggtcgttg aaacaaggtg gggggcatgg tgggcggcaa gaacccaagg
     3901 tcttgagccc ttcgctaatg cgggaaagct cttattcggg tgagatgggc tgggcaccat
     3961 ctggggaccc tgacgtgaag tttgtcactg actggagaac tcggtttgtc gtctgttgcg
     4021 ggggcggcag ttatggcggt gccgttgggc agtgcacccg tacctttggg agcgcgcgcc
     4081 ctcgtcgtgt cgtgacgtca cccgttctgt tggcttataa tgcagggtgg ggccaccagc
     4141 cggtaggtgt gcggtaggct tttctccgtc gcaggacgca gggttcgggc ctagggtagg
     4201 ctctcctgaa tcgacaggcg ccggacctct ggtgagggga gggataagtg aggcgtcagt
     4261 ttctttggtc ggttttatgt acctatcttc ttaagtagct gaagctccgg ttttgaacta
     4321 tgcgctcggg gttggcgagt gtgttttgtg aagtttttta ggcacctttt gaaatgtaat
     4381 catttgggtc aatatgtaat tttcagtgtt agacttgtaa attgtccgct aaattctggc
     4441 cgtttttggc ttttttgtta gacaacatgg gtaaaaagcc tgaactcacc gcgacgtctg
     4501 tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg
     4561 gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa
     4621 atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatccgccg
     4681 cgctcccgat tccggaagtg cttgacattg gggagttcag cgagagcctg acctattgca
     4741 tctcccgccg tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg
     4801 ttctgcagcc ggtcgcggag gcaatggatg ccatcgctgc cgccgatctt agccagacga
     4861 gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca
     4921 tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca
     4981 gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag
     5041 tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca
     5101 taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca
     5161 acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc
     5221 ggaggcatcc ggagcttgca ggatcgcccc ggctccgggc gtatatgctc cgcattggtc
     5281 ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg
     5341 gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc
     5401 gcagaagcgc cgccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc
     5461 gacgccccag cactcgtccg agggcaaagg aataagctag tatgtaagcc tagtcttaga
     5521 taataaaatc gctatccatc gaagatggat gtgtgttggt tttttgtgtg tgtaacgcta
     5581 ggcgcgcctg gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt
     5641 gcttggcagt cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat
     5701 tgtactcaag cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct
     5761 gatcggtcaa cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca
     5821 gcaggaggct ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5881 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5941 gatcagttgg gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     6001 gagagttttc gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt
     6061 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     6121 tctcagaatg acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg
     6181 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     6241 cttctgacaa cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     6301 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     6361 cgtgacacca cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa
     6421 ctacttactc tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca
     6481 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     6541 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     6601 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     6661 gctgagatag gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag
     6721 aaaaaaatgc ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa
     6781 cggatacggc ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc
     6841 cttaagatcc cgaat
//
