LOCUS       sequence-307254-        4989 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 4989)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4989)
  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-m4133408
FEATURES             Location/Qualifiers
     source          1..4989
                     /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             862..939
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     CDS             967..987
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1436..1557
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1447..1466)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1501..1520
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1672..1718
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1748..1884
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1929..2000
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2226..2814)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2306..2325)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3560..3628,3629..4420)
                     /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(3778..3797)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4433..4569
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4721..4792
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4797..4843)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4861..4909
                     /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 ccatgtactc attcgtttcg
      601 gaagagacag gtacgttaat agttaatagc gtacttcttt ttcttgcttt cgtggtattc
      661 ttgctagtta cactagccat ccttactgcg cttcgattgt gtgcgtactg ctgcaatatt
      721 gttaacgtga gtcttgtaaa accttctttt tacgtttact ctcgtgttaa aaatctgaat
      781 tcttctagag ttcctgatct tctggtcgaa aacctttact ttcagtcggg acggcgaatc
      841 ccggggctga taaatccctg gaaaaggaga tggaagaaga actttattgc agtgtctgcg
      901 gctaaccggt tcaaaaagat cagttcaagc ggcgccctag attacgacat tcctacgacc
      961 gcctccgaga atctctattt ccagggtgaa tttggcttgg cacaacatga cgaggccgtc
     1021 gataacaagt ttaataaaga acagcagaac gctttctatg agatccttca cttaccaaat
     1081 ctgaacgaag agcaacgcaa cgctttcatt cagagcctga aagacgatcc ctcgcagtct
     1141 gcaaatctcc tggccgaagc caagaagctg aatgacgcac aggctccaaa agttgataac
     1201 aagtttaaca aggagcagca gaatgctttc tacgagatat tgcaccttcc taatctgaat
     1261 gaggaacaaa gaaatgcatt tatccagtcc ctaaaagacg atccatcaca gagtgccaac
     1321 ctgcttgcgg aagccaaaaa gctcaatgat gcccaggccc cgaaagtaga cgcgaaccat
     1381 caataagcta gcttgactga ctgagataca gcgtaccttc agctcacaga catgataaga
     1441 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     1501 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     1561 aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa
     1621 agcaagtaaa acctctacaa atgtggtatt ggcccatctc tatcggtatc gtagcataac
     1681 cccttggggc ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc cggattgcta
     1741 tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca
     1801 catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc
     1861 tccttaccag aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc tctcgatttc
     1921 gatcaagaca ttcctttaat ggtcttttct ggacaccact aggggtcaga agtagttcat
     1981 caaactttct tccctcccta atctcattgg ttaccttggg ctatcgaaac ttaattaacc
     2041 agtcaagtca gctacttggc gagatcgact tgtctgggtt tcgactacgc tcagaattgc
     2101 gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt ctccgattac gagtttcatt
     2161 taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2221 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2281 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2341 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2401 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2461 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2521 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2581 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2641 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2701 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2761 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2821 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     2881 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2941 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3001 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc atttaaattt
     3061 ccgaactctc caaggccctc gtcggaaaat cttcaaacct ttcgtccgat ccatcttgca
     3121 ggctacctct cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct tggctattgc
     3181 ttggcagcgc ctatcgccag gtattactcc aatcccgaat atccgagatc gggatcaccc
     3241 gagagaagtt caacctacat cctcaatccc gatctatccg agatccgagg aatatcgaaa
     3301 tcggggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga cgatccatat
     3361 cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc caatcgtcag
     3421 atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta gatattgata
     3481 gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc aagagacagg
     3541 atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta ttcccttttt
     3601 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     3661 tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca gcggtaagat
     3721 ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta aagttctgct
     3781 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     3841 ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc ttacggatgg
     3901 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     3961 cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4021 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4081 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg
     4141 cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg cggataaagt
     4201 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     4261 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4321 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     4381 gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag gtgcattggc
     4441 gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca
     4501 gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt
     4561 tatccttaag atcccgaatc gtttaaactc gactctggct ctatcgaatc tccgtcgttt
     4621 cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct
     4681 atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac cattagctcc
     4741 acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc tcaattcaaa
     4801 aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg ttgcgttaca
     4861 cacacaaaaa accaacacac atccatcttc gatggatagc gattttatta tctaactgct
     4921 gatcgagtgt agccagatct agtaatcaat tacggggtca ttagttcata gcccatatat
     4981 ggagttccg
//
