LOCUS       sequence-295896-        7775 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 7775)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7775)
  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-m4133713
FEATURES             Location/Qualifiers
     source          1..7775
                     /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             725..802
                     /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             830..850
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1315..1436
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1326..1345)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1380..1399
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1551..1597
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1627..1763
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1808..1879
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2105..2693)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2185..2204)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3193..4402
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4239..4258
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4406..5434
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5461..5509
                     /note="synthetic polyadenylation signal"
     CDS             join(5775..5843,5844..6635)
                     /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(5993..6012)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6648..6784
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6929..7000
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7005..7051)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7069..7117
                     /note="synthetic polyadenylation signal"
     enhancer        7198..7501
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7503..7705
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7656..7676
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(7774..7775,1..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 gggacggcga atcccggggc tgataaatcc
      721 ctggaaaagg agatggaaga agaactttat tgcagtgtct gcggctaacc ggttcaaaaa
      781 gatcagttca agcggcgccc tagattacga cattcctacg accgcctccg agaatctcta
      841 tttccagggt gaatttggct tggcacaaca tgacgaggcc gtcgataaca agtttaataa
      901 agaacagcag aacgctttct atgagatcct tcacttacca aatctgaacg aagagcaacg
      961 caacgctttc attcagagcc tgaaagacga tccctcgcag tctgcaaatc tcctggccga
     1021 agccaagaag ctgaatgacg cacaggctcc aaaagttgat aacaagttta acaaggagca
     1081 gcagaatgct ttctacgaga tattgcacct tcctaatctg aatgaggaac aaagaaatgc
     1141 atttatccag tccctaaaag acgatccatc acagagtgcc aacctgcttg cggaagccaa
     1201 aaagctcaat gatgcccagg ccccgaaagt agacgcgaac catcaataag gatccgcagg
     1261 cctctgctag cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat
     1321 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     1381 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     1441 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
     1501 gcaagtaaaa cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc
     1561 ccttggggcc tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat
     1621 ctaccggcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac
     1681 atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct
     1741 ccttaccaga aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg
     1801 atcaagacat tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc
     1861 aaactttctt ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca
     1921 gtcaagtcag ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg
     1981 tcagtcaagt tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt
     2041 aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2101 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2161 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2221 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2281 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2341 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2401 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2461 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2521 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     2581 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     2641 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     2701 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     2761 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     2821 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     2881 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc
     2941 cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag
     3001 gctacctctc gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct
     3061 tggcagcgcc tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg
     3121 agagaagttc aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat
     3181 cggggcgcgc ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc cccctcgtca
     3241 cggcgagcgc tgccacgtca gacgaagggc gcaggagcgt cctgatcctt ccgcccggac
     3301 gctcaggaca gcggcccgct gctcataaga ctcggcctta gaaccccagt atcagcagaa
     3361 ggacatttta ggacgggact tgggtgactc tagggcactg gttttctttc cagagagcgg
     3421 aacaggcgag gaaaagtagt cccttctcgg cgattctgcg gagggatctc cgtggggcgg
     3481 tgaacgccga tgattatata aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc
     3541 gggatttggg tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg
     3601 gctgctgggc tggccggggc tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt
     3661 ggagagaccg ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg aactgggggt
     3721 tggggggagc gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag
     3781 gcgggctgtg aggtcgttga aacaaggtgg ggggcatggt gggcggcaag aacccaaggt
     3841 cttgagccct tcgctaatgc gggaaagctc ttattcgggt gagatgggct gggcaccatc
     3901 tggggaccct gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg
     3961 gggcggcagt tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc
     4021 tcgtcgtgtc gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccacctgcc
     4081 ggtaggtgtg cggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc
     4141 tctcctgaat cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt
     4201 tctttggtcg gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat
     4261 gcgctcgggg ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc
     4321 atttgggtca atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc
     4381 gtttttggct tttttgttag acaacatggg taaaaagcct gaactcaccg cgacgtctgt
     4441 cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg
     4501 cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa
     4561 tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg catccgccgc
     4621 gctcccgatt ccggaagtgc ttgacattgg ggagttcagc gagagcctga cctattgcat
     4681 ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt
     4741 tctgcagccg gtcgcggagg caatggatgc catcgctgcc gccgatctta gccagacgag
     4801 cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat
     4861 atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag
     4921 tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt
     4981 ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat
     5041 aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa
     5101 catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg
     5161 gaggcatccg gagcttgcag gatcgccccg gctccgggcg tatatgctcc gcattggtct
     5221 tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg
     5281 tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg
     5341 cagaagcgcc gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg
     5401 acgccccagc actcgtccga gggcaaagga ataagctagt atgtaagcct agtcttagat
     5461 aataaaatcg ctatccatcg aagatggatg tgtgttggtt ttttgtgtgt gtaacgctag
     5521 gcgcgcctgg tgtaccgaga acgatcctct cagtgcgagt ctcgacgatc catatcgttg
     5581 cttggcagtc agccagtcgg aatccagctt gggacccagg aagtccaatc gtcagatatt
     5641 gtactcaagc ctggtcacgg cagcgtaccg atctgtttaa acctagatat tgatagtctg
     5701 atcggtcaac gtataatcga gtcctagctt ttgcaaacat ctatcaagag acaggatcag
     5761 caggaggctt tcgcatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5821 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5881 atcagttggg tgcgcgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5941 agagttttcg ccccgaagaa cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg
     6001 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     6061 ctcagaatga cttggttgag tattcaccag tcacagaaaa gcatcttacg gatggcatga
     6121 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     6181 ttctgacaac gattggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     6241 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     6301 gtgacaccac gatgcctgta gcaatggcaa caaccttgcg taaactatta actggcgaac
     6361 tacttactct agcttcccgg caacagttga tagactggat ggaggcggat aaagttgcag
     6421 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     6481 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     6541 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     6601 ctgagatagg tgcctcactg attaagcatt ggtaaccgat tctaggtgca ttggcgcaga
     6661 aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac
     6721 ggatacggct tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc
     6781 ttaagatcgt ttaaactcga ctctggctct atcgaatctc cgtcgtttcg agcttacgcg
     6841 aacagccgtg gcgctcattt gctcgtcggg catcgaatct cgtcagctat cgtcagctta
     6901 cctttttggc agcgatcgcg gctcccgaca tcttggacca ttagctccac aggtatcttc
     6961 ttccctctag tggtcataac agcagcttca gctacctctc aattcaaaaa acccctcaag
     7021 acccgtttag aggccccaag gggttatgct atcaatcgtt gcgttacaca cacaaaaaac
     7081 caacacacat ccatcttcga tggatagcga ttttattatc taactgctga tcgagtgtag
     7141 ccagatctag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     7201 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     7261 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     7321 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     7381 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     7441 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     7501 gctgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     7561 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     7621 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     7681 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatcagat ctttgtcgat
     7741 cctaccatcc actcgacaca cccgccagcg gccgc
//
