LOCUS       sequence-295652-        7247 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 7247)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7247)
  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-m4134549
FEATURES             Location/Qualifiers
     source          1..7247
                     /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..718
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    787..908
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(798..817)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     852..871
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1023..1069
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1099..1235
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1280..1351
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1577..2165)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1657..1676)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2665..3874
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3711..3730
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3878..4906
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4933..4981
                     /note="synthetic polyadenylation signal"
     CDS             join(5247..5315,5316..6107)
                     /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(5465..5484)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6120..6256
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6401..6472
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6477..6523)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6541..6589
                     /note="synthetic polyadenylation signal"
     enhancer        6670..6973
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6975..7177
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7128..7148
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(7246..7247,1..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 gtacccctat gacgtaccag attatgcgta
      721 aggatccgca ggcctctgct agcttgactg actgagatac agcgtacctt cagctcacag
      781 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
      841 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
      901 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
      961 aggtttttta aagcaagtaa aacctctaca aatgtggtat tggcccatct ctatcggtat
     1021 cgtagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgt gcccctcggg
     1081 ccggattgct atctaccggc attggcgcag aaaaaaatgc ctgatgcgac gctgcgcgtc
     1141 ttatactccc acatatgcca gattcagcaa cggatacggc ttccccaact tgcccacttc
     1201 catacgtgtc ctccttacca gaaatttatc cttaaggtcg tcagctatcc tgcaggcgat
     1261 ctctcgattt cgatcaagac attcctttaa tggtcttttc tggacaccac taggggtcag
     1321 aagtagttca tcaaactttc ttccctccct aatctcattg gttaccttgg gctatcgaaa
     1381 cttaattaac cagtcaagtc agctacttgg cgagatcgac ttgtctgggt ttcgactacg
     1441 ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta ttgcacccgt tctccgatta
     1501 cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     1561 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     1621 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     1681 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     1741 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1801 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1861 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1921 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     1981 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     2041 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     2101 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     2161 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     2221 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     2281 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     2341 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     2401 catttaaatt tccgaactct ccaaggccct cgtcggaaaa tcttcaaacc tttcgtccga
     2461 tccatcttgc aggctacctc tcgaacgaac tatcgcaagt ctcttggccg gccttgcgcc
     2521 ttggctattg cttggcagcg cctatcgcca ggtattactc caatcccgaa tatccgagat
     2581 cgggatcacc cgagagaagt tcaacctaca tcctcaatcc cgatctatcc gagatccgag
     2641 gaatatcgaa atcggggcgc gcctggcctc cgcgccgggt tttggcgcct cccgcgggcg
     2701 cccccctcgt cacggcgagc gctgccacgt cagacgaagg gcgcaggagc gtcctgatcc
     2761 ttccgcccgg acgctcagga cagcggcccg ctgctcataa gactcggcct tagaacccca
     2821 gtatcagcag aaggacattt taggacggga cttgggtgac tctagggcac tggttttctt
     2881 tccagagagc ggaacaggcg aggaaaagta gtcccttctc ggcgattctg cggagggatc
     2941 tccgtggggc ggtgaacgcc gatgattata taaggacgcg ccgggtgtgg cacagctagt
     3001 tccgtcgcag ccgggatttg ggtcgcggtt cttgtttgtg gatcgctgtg atcgtcactt
     3061 ggtgagtagc gggctgctgg gctggccggg gctttcgtgg ccgccgggcc gctcggtggg
     3121 acggaagcgt gtggagagac cgccaagggc tgtagtctgg gtccgcgagc aaggttgccc
     3181 tgaactgggg gttgggggga gcgcagcaaa atggcggctg ttcccgagtc ttgaatggaa
     3241 gacgcttgtg aggcgggctg tgaggtcgtt gaaacaaggt ggggggcatg gtgggcggca
     3301 agaacccaag gtcttgagcc cttcgctaat gcgggaaagc tcttattcgg gtgagatggg
     3361 ctgggcacca tctggggacc ctgacgtgaa gtttgtcact gactggagaa ctcggtttgt
     3421 cgtctgttgc gggggcggca gttatggcgg tgccgttggg cagtgcaccc gtacctttgg
     3481 gagcgcgcgc cctcgtcgtg tcgtgacgtc acccgttctg ttggcttata atgcagggtg
     3541 gggccacctg ccggtaggtg tgcggtaggc ttttctccgt cgcaggacgc agggttcggg
     3601 cctagggtag gctctcctga atcgacaggc gccggacctc tggtgagggg agggataagt
     3661 gaggcgtcag tttctttggt cggttttatg tacctatctt cttaagtagc tgaagctccg
     3721 gttttgaact atgcgctcgg ggttggcgag tgtgttttgt gaagtttttt aggcaccttt
     3781 tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt tagacttgta aattgtccgc
     3841 taaattctgg ccgtttttgg cttttttgtt agacaacatg ggtaaaaagc ctgaactcac
     3901 cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca
     3961 gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt
     4021 cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt
     4081 tgcatccgcc gcgctcccga ttccggaagt gcttgacatt ggggagttca gcgagagcct
     4141 gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga
     4201 actgcccgct gttctgcagc cggtcgcgga ggcaatggat gccatcgctg ccgccgatct
     4261 tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg
     4321 gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga
     4381 cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga
     4441 ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga
     4501 caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata
     4561 cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg
     4621 ctacttcgag cggaggcatc cggagcttgc aggatcgccc cggctccggg cgtatatgct
     4681 ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc
     4741 ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac
     4801 acaaatcgcc cgcagaagcg ccgccgtctg gaccgatggc tgtgtagaag tactcgccga
     4861 tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaataagcta gtatgtaagc
     4921 ctagtcttag ataataaaat cgctatccat cgaagatgga tgtgtgttgg ttttttgtgt
     4981 gtgtaacgct aggcgcgcct ggtgtaccga gaacgatcct ctcagtgcga gtctcgacga
     5041 tccatatcgt tgcttggcag tcagccagtc ggaatccagc ttgggaccca ggaagtccaa
     5101 tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac cgatctgttt aaacctagat
     5161 attgatagtc tgatcggtca acgtataatc gagtcctagc ttttgcaaac atctatcaag
     5221 agacaggatc agcaggaggc tttcgcatga gtattcaaca tttccgtgtc gcccttattc
     5281 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     5341 aagatgctga agatcagttg ggtgcgcgag tgggttacat cgaactggat ctcaacagcg
     5401 gtaagatcct tgagagtttt cgccccgaag aacgctttcc aatgatgagc acttttaaag
     5461 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     5521 gcatacacta ttctcagaat gacttggttg agtattcacc agtcacagaa aagcatctta
     5581 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     5641 cggccaactt acttctgaca acgattggag gaccgaagga gctaaccgct tttttgcaca
     5701 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     5761 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaaccttg cgtaaactat
     5821 taactggcga actacttact ctagcttccc ggcaacagtt gatagactgg atggaggcgg
     5881 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     5941 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     6001 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     6061 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaaccg attctaggtg
     6121 cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc
     6181 agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc
     6241 agaaatttat ccttaagatc gtttaaactc gactctggct ctatcgaatc tccgtcgttt
     6301 cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct
     6361 atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac cattagctcc
     6421 acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc tcaattcaaa
     6481 aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg ttgcgttaca
     6541 cacacaaaaa accaacacac atccatcttc gatggatagc gattttatta tctaactgct
     6601 gatcgagtgt agccagatct agtaatcaat tacggggtca ttagttcata gcccatatat
     6661 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     6721 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     6781 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     6841 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     6901 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     6961 cgctattacc atgctgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     7021 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     7081 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     7141 taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatcag
     7201 atctttgtcg atcctaccat ccactcgaca cacccgccag cggccgc
//
