LOCUS       sequence-295832-        4917 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid for SARS-CoV-2 M (membrane).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4917)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4917)
  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-m4133585
FEATURES             Location/Qualifiers
     source          1..4917
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             671..691
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             692..733
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    786..907
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(797..816)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     851..870
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1022..1068
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1098..1234
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1279..1350
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1576..2164)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1656..1675)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2910..2978,2979..3770)
                     /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(3128..3147)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3783..3919
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4071..4142
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4147..4193)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4211..4259
                     /note="synthetic polyadenylation signal"
     enhancer        4340..4643
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4645..4847
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4798..4818
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(4916..4917,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggca gattccaacg gtactattac cgttgaagag cttaaaaagc tccttgaaca
       61 atggaaccta gtaataggtt tcctattcct tacatggatt tgtcttctac aatttgccta
      121 tgccaacagg aataggtttt tgtatataat taagttaatt ttcctctggc tgttatggcc
      181 agtaacttta gcttgttttg tgcttgctgc tgtttacaga ataaattgga tcaccggtgg
      241 aattgctatc gcaatggctt gtcttgtagg cttgatgtgg ctcagctact tcattgcttc
      301 tttcagactg tttgcgcgta cgcgttccat gtggtcattc aatccagaaa ctaacattct
      361 tctcaacgtg ccactccatg gcactattct gaccagaccg cttctagaaa gtgaactcgt
      421 aatcggagct gtgatccttc gtggacatct tcgtattgct ggacaccatc taggacgctg
      481 tgacatcaag gacctgccta aagaaatcac tgttgctaca tcacgaacgc tttcttatta
      541 caaattggga gcttcgcagc gtgtagcagg tgactcaggt tttgctgcat acagtcgcta
      601 caggattggc aactataaat taaacacaga ccattccagt agcagtgaca atattgcttt
      661 gcttgtacag gaaaaccttt actttcagtc ggggaaacct atacccaatc cacttctcgg
      721 cttagattct acctaagcta gcttgactga ctgagataca gcgtaccttc agctcacaga
      781 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
      841 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
      901 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
      961 ggttttttaa agcaagtaaa acctctacaa atgtggtatt ggcccatctc tatcggtatc
     1021 gtagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgtg cccctcgggc
     1081 cggattgcta tctaccggca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
     1141 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
     1201 atacgtgtcc tccttaccag aaatttatcc ttaaggtcgt cagctatcct gcaggcgatc
     1261 tctcgatttc gatcaagaca ttcctttaat ggtcttttct ggacaccact aggggtcaga
     1321 agtagttcat caaactttct tccctcccta atctcattgg ttaccttggg ctatcgaaac
     1381 ttaattaacc agtcaagtca gctacttggc gagatcgact tgtctgggtt tcgactacgc
     1441 tcagaattgc gtcagtcaag ttcgatctgg tccttgctat tgcacccgtt ctccgattac
     1501 gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1561 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1621 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1681 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1741 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1801 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1861 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1921 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1981 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2041 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2101 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2161 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2221 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2281 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     2341 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     2401 atttaaattt ccgaactctc caaggccctc gtcggaaaat cttcaaacct ttcgtccgat
     2461 ccatcttgca ggctacctct cgaacgaact atcgcaagtc tcttggccgg ccttgcgcct
     2521 tggctattgc ttggcagcgc ctatcgccag gtattactcc aatcccgaat atccgagatc
     2581 gggatcaccc gagagaagtt caacctacat cctcaatccc gatctatccg agatccgagg
     2641 aatatcgaaa tcggggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga
     2701 cgatccatat cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc
     2761 caatcgtcag atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta
     2821 gatattgata gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc
     2881 aagagacagg atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta
     2941 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     3001 taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca
     3061 gcggtaagat ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta
     3121 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     3181 gccgcataca ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc
     3241 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     3301 ctgcggccaa cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc
     3361 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     3421 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac
     3481 tattaactgg cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg
     3541 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     3601 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     3661 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     3721 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag
     3781 gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
     3841 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
     3901 accagaaatt tatccttaag atcccgaatc gtttaaactc gactctggct ctatcgaatc
     3961 tccgtcgttt cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat
     4021 ctcgtcagct atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac
     4081 cattagctcc acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc
     4141 tcaattcaaa aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg
     4201 ttgcgttaca cacacaaaaa accaacacac atccatcttc gatggatagc gattttatta
     4261 tctaactgct gatcgagtgt agccagatct agtaatcaat tacggggtca ttagttcata
     4321 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     4381 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     4441 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     4501 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     4561 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     4621 tattagtcat cgctattacc atgctgatgc ggttttggca gtacatcaat gggcgtggat
     4681 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     4741 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     4801 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
     4861 gtcagatcag atctttgtcg atcctaccat ccactcgaca cacccgccag cggccgc
//
