LOCUS       sequence-295679-        7265 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 7265)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7265)
  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-m4134302
FEATURES             Location/Qualifiers
     source          1..7265
                     /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..736
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    805..926
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(816..835)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     870..889
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1041..1087
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1117..1253
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1298..1369
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1595..2183)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1675..1694)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2683..3892
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3729..3748
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3896..4924
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4951..4999
                     /note="synthetic polyadenylation signal"
     CDS             join(5265..5333,5334..6125)
                     /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(5483..5502)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6138..6274
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6419..6490
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6495..6541)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6559..6607
                     /note="synthetic polyadenylation signal"
     enhancer        6688..6991
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6993..7195
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7146..7166
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(7264..7265,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggcg gacagcaacg gcactattac agtcgaagag cttaagaaat tactcgagca
       61 gtggaatttg gtaataggat tcctgtttct aacgtggatc tgtctgctgc aattcgcata
      121 tgccaaccgg aatagatttc tctacatcat taagttgatc ttcctgtggc tgctttggcc
      181 tgtgaccttg gcttgctttg ttctggccgc cgtgtaccgc ataaactgga ttaccggtgg
      241 gatcgctatc gcaatggcgt gcctcgtggg ccttatgtgg ctgtcctatt ttatagcctc
      301 attccgactc tttgctcgta ctaggagtat gtggtctttc aatcccgaaa caaatatttt
      361 actaaacgtc ccgctgcatg ggaccatttt aacaagacca ttgctagaga gcgagctcgt
      421 gatcggcgca gttatactgc ggggacacct caggatcgcc ggacaccatc tgggtcgctg
      481 tgatattaaa gacctgccca aggaaataac agtggctacc tcgcgtactc tttcatacta
      541 taaactggga gccagccaga gagtggccgg ggattctggc ttcgcagcat atagtcgata
      601 caggatcggg aactacaagc tgaatacgga ccactcctcc tctagtgata atattgcatt
      661 gctcgtacag gaaaaccttt actttcagtc gaaagaaacc gcggcagcta agttcgagag
      721 gcagcacatg gacagttaag gatccgcagg cctctgctag cttgactgac tgagatacag
      781 cgtaccttca gctcacagac atgataagat acattgatga gtttggacaa accacaacta
      841 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
      901 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
      961 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg
     1021 gcccatctct atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg
     1081 ttttttgtgc ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct
     1141 gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt
     1201 ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc
     1261 agctatcctg caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg
     1321 gacaccacta ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt
     1381 taccttgggc tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt
     1441 gtctgggttt cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt
     1501 gcacccgttc tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg
     1561 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     1621 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     1681 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     1741 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     1801 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     1861 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     1921 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     1981 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     2041 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     2101 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     2161 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     2221 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     2281 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     2341 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     2401 ttcgttcatc catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc
     2461 ttcaaacctt tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct
     2521 cttggccggc cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca
     2581 atcccgaata tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg
     2641 atctatccga gatccgagga atatcgaaat cggggcgcgc ctggcctccg cgccgggttt
     2701 tggcgcctcc cgcgggcgcc cccctcgtca cggcgagcgc tgccacgtca gacgaagggc
     2761 gcaggagcgt cctgatcctt ccgcccggac gctcaggaca gcggcccgct gctcataaga
     2821 ctcggcctta gaaccccagt atcagcagaa ggacatttta ggacgggact tgggtgactc
     2881 tagggcactg gttttctttc cagagagcgg aacaggcgag gaaaagtagt cccttctcgg
     2941 cgattctgcg gagggatctc cgtggggcgg tgaacgccga tgattatata aggacgcgcc
     3001 gggtgtggca cagctagttc cgtcgcagcc gggatttggg tcgcggttct tgtttgtgga
     3061 tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc tggccggggc tttcgtggcc
     3121 gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg ccaagggctg tagtctgggt
     3181 ccgcgagcaa ggttgccctg aactgggggt tggggggagc gcagcaaaat ggcggctgtt
     3241 cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg aggtcgttga aacaaggtgg
     3301 ggggcatggt gggcggcaag aacccaaggt cttgagccct tcgctaatgc gggaaagctc
     3361 ttattcgggt gagatgggct gggcaccatc tggggaccct gacgtgaagt ttgtcactga
     3421 ctggagaact cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg ccgttgggca
     3481 gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac ccgttctgtt
     3541 ggcttataat gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt ttctccgtcg
     3601 caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc cggacctctg
     3661 gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta cctatcttct
     3721 taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg tgttttgtga
     3781 agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta
     3841 gacttgtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag acaacatggg
     3901 taaaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag
     3961 cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt
     4021 aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg
     4081 ttatgtttat cggcactttg catccgccgc gctcccgatt ccggaagtgc ttgacattgg
     4141 ggagttcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca
     4201 agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg caatggatgc
     4261 catcgctgcc gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat
     4321 cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca
     4381 ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct
     4441 gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc
     4501 caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat
     4561 gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg
     4621 tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccccg
     4681 gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg
     4741 caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc
     4801 cgggactgtc gggcgtacac aaatcgcccg cagaagcgcc gccgtctgga ccgatggctg
     4861 tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga
     4921 ataagctagt atgtaagcct agtcttagat aataaaatcg ctatccatcg aagatggatg
     4981 tgtgttggtt ttttgtgtgt gtaacgctag gcgcgcctgg tgtaccgaga acgatcctct
     5041 cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt
     5101 gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg
     5161 atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt
     5221 ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgagt attcaacatt
     5281 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     5341 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg
     5401 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa
     5461 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     5521 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag
     5581 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5641 ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc
     5701 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5761 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5821 caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg caacagttga
     5881 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5941 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     6001 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     6061 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     6121 ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct
     6181 tatactccca catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc
     6241 atacgtgtcc tccttaccag aaatttatcc ttaagatcgt ttaaactcga ctctggctct
     6301 atcgaatctc cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt gctcgtcggg
     6361 catcgaatct cgtcagctat cgtcagctta cctttttggc agcgatcgcg gctcccgaca
     6421 tcttggacca ttagctccac aggtatcttc ttccctctag tggtcataac agcagcttca
     6481 gctacctctc aattcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct
     6541 atcaatcgtt gcgttacaca cacaaaaaac caacacacat ccatcttcga tggatagcga
     6601 ttttattatc taactgctga tcgagtgtag ccagatctag taatcaatta cggggtcatt
     6661 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     6721 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     6781 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     6841 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     6901 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     6961 catctacgta ttagtcatcg ctattaccat gctgatgcgg ttttggcagt acatcaatgg
     7021 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     7081 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     7141 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
     7201 agtgaaccgt cagatcagat ctttgtcgat cctaccatcc actcgacaca cccgccagcg
     7261 gccgc
//
