LOCUS       sequence-335172-        7296 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 7296)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7296)
  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-m4133645
FEATURES             Location/Qualifiers
     source          1..7296
                     /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"
     regulatory      577..586
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1249..1269
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1273..1338
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1407..1528
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1418..1437)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1472..1491
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1643..1689
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1719..1855
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1900..1971
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2197..2785)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2277..2296)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3285..4494
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4331..4350
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4498..5526
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5553..5601
                     /note="synthetic polyadenylation signal"
     CDS             join(5867..5935,5936..6727)
                     /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(6085..6104)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6740..6876
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7028..7099
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7104..7150)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7168..7216
                     /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 ccatggcaga ttccaacggt
      601 actattaccg ttgaagagct taaaaagctc cttgaacaat ggaacctagt aataggtttc
      661 ctattcctta catggatttg tcttctacaa tttgcctatg ccaacaggaa taggtttttg
      721 tatataatta agttaatttt cctctggctg ttatggccag taactttagc ttgttttgtg
      781 cttgctgctg tttacagaat aaattggatc accggtggaa ttgctatcgc aatggcttgt
      841 cttgtaggct tgatgtggct cagctacttc attgcttctt tcagactgtt tgcgcgtacg
      901 cgttccatgt ggtcattcaa tccagaaact aacattcttc tcaacgtgcc actccatggc
      961 actattctga ccagaccgct tctagaaagt gaactcgtaa tcggagctgt gatccttcgt
     1021 ggacatcttc gtattgctgg acaccatcta ggacgctgtg acatcaagga cctgcctaaa
     1081 gaaatcactg ttgctacatc acgaacgctt tcttattaca aattgggagc ttcgcagcgt
     1141 gtagcaggtg actcaggttt tgctgcatac agtcgctaca ggattggcaa ctataaatta
     1201 aacacagacc attccagtag cagtgacaat attgctttgc ttgtacagga aaacctttac
     1261 tttcagtcga tggattataa ggaccacgat ggagactaca aagatcatga cattgattac
     1321 aaggacgacg atgacaaata aggatccgca ggcctctgct agcttgactg actgagatac
     1381 agcgtacctt cagctcacag acatgataag atacattgat gagtttggac aaaccacaac
     1441 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     1501 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     1561 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat
     1621 tggcccatct ctatcggtat cgtagcataa ccccttgggg cctctaaacg ggtcttgagg
     1681 ggttttttgt gcccctcggg ccggattgct atctaccggc attggcgcag aaaaaaatgc
     1741 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc
     1801 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaaggtcg
     1861 tcagctatcc tgcaggcgat ctctcgattt cgatcaagac attcctttaa tggtcttttc
     1921 tggacaccac taggggtcag aagtagttca tcaaactttc ttccctccct aatctcattg
     1981 gttaccttgg gctatcgaaa cttaattaac cagtcaagtc agctacttgg cgagatcgac
     2041 ttgtctgggt ttcgactacg ctcagaattg cgtcagtcaa gttcgatctg gtccttgcta
     2101 ttgcacccgt tctccgatta cgagtttcat ttaaatcatg tgagcaaaag gccagcaaaa
     2161 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2221 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2281 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2341 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2401 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2461 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2521 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2581 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2641 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2701 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2761 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2821 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2881 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     2941 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3001 atttcgttca tccatagttg catttaaatt tccgaactct ccaaggccct cgtcggaaaa
     3061 tcttcaaacc tttcgtccga tccatcttgc aggctacctc tcgaacgaac tatcgcaagt
     3121 ctcttggccg gccttgcgcc ttggctattg cttggcagcg cctatcgcca ggtattactc
     3181 caatcccgaa tatccgagat cgggatcacc cgagagaagt tcaacctaca tcctcaatcc
     3241 cgatctatcc gagatccgag gaatatcgaa atcggggcgc gcctggcctc cgcgccgggt
     3301 tttggcgcct cccgcgggcg cccccctcgt cacggcgagc gctgccacgt cagacgaagg
     3361 gcgcaggagc gtcctgatcc ttccgcccgg acgctcagga cagcggcccg ctgctcataa
     3421 gactcggcct tagaacccca gtatcagcag aaggacattt taggacggga cttgggtgac
     3481 tctagggcac tggttttctt tccagagagc ggaacaggcg aggaaaagta gtcccttctc
     3541 ggcgattctg cggagggatc tccgtggggc ggtgaacgcc gatgattata taaggacgcg
     3601 ccgggtgtgg cacagctagt tccgtcgcag ccgggatttg ggtcgcggtt cttgtttgtg
     3661 gatcgctgtg atcgtcactt ggtgagtagc gggctgctgg gctggccggg gctttcgtgg
     3721 ccgccgggcc gctcggtggg acggaagcgt gtggagagac cgccaagggc tgtagtctgg
     3781 gtccgcgagc aaggttgccc tgaactgggg gttgggggga gcgcagcaaa atggcggctg
     3841 ttcccgagtc ttgaatggaa gacgcttgtg aggcgggctg tgaggtcgtt gaaacaaggt
     3901 ggggggcatg gtgggcggca agaacccaag gtcttgagcc cttcgctaat gcgggaaagc
     3961 tcttattcgg gtgagatggg ctgggcacca tctggggacc ctgacgtgaa gtttgtcact
     4021 gactggagaa ctcggtttgt cgtctgttgc gggggcggca gttatggcgg tgccgttggg
     4081 cagtgcaccc gtacctttgg gagcgcgcgc cctcgtcgtg tcgtgacgtc acccgttctg
     4141 ttggcttata atgcagggtg gggccacctg ccggtaggtg tgcggtaggc ttttctccgt
     4201 cgcaggacgc agggttcggg cctagggtag gctctcctga atcgacaggc gccggacctc
     4261 tggtgagggg agggataagt gaggcgtcag tttctttggt cggttttatg tacctatctt
     4321 cttaagtagc tgaagctccg gttttgaact atgcgctcgg ggttggcgag tgtgttttgt
     4381 gaagtttttt aggcaccttt tgaaatgtaa tcatttgggt caatatgtaa ttttcagtgt
     4441 tagacttgta aattgtccgc taaattctgg ccgtttttgg cttttttgtt agacaacatg
     4501 ggtaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac
     4561 agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat
     4621 gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat
     4681 cgttatgttt atcggcactt tgcatccgcc gcgctcccga ttccggaagt gcttgacatt
     4741 ggggagttca gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg
     4801 caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggcaatggat
     4861 gccatcgctg ccgccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga
     4921 atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat
     4981 cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag
     5041 ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc
     5101 tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg
     5161 atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct
     5221 tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccc
     5281 cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac
     5341 ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga
     5401 gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg ccgccgtctg gaccgatggc
     5461 tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag
     5521 gaataagcta gtatgtaagc ctagtcttag ataataaaat cgctatccat cgaagatgga
     5581 tgtgtgttgg ttttttgtgt gtgtaacgct aggcgcgcct ggtgtaccga gaacgatcct
     5641 ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc ggaatccagc
     5701 ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac
     5761 cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc gagtcctagc
     5821 ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgcatga gtattcaaca
     5881 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     5941 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcgcgag tgggttacat
     6001 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgctttcc
     6061 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     6121 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtattcacc
     6181 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     6241 aaccatgagt gataacactg cggccaactt acttctgaca acgattggag gaccgaagga
     6301 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     6361 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     6421 aacaaccttg cgtaaactat taactggcga actacttact ctagcttccc ggcaacagtt
     6481 gatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     6541 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     6601 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     6661 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     6721 ttggtaaccg attctaggtg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt
     6781 cttatactcc cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt
     6841 ccatacgtgt cctccttacc agaaatttat ccttaagatc ccgaatcgtt taaactcgac
     6901 tctggctcta tcgaatctcc gtcgtttcga gcttacgcga acagccgtgg cgctcatttg
     6961 ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac ctttttggca gcgatcgcgg
     7021 ctcccgacat cttggaccat tagctccaca ggtatcttct tccctctagt ggtcataaca
     7081 gcagcttcag ctacctctca attcaaaaaa cccctcaaga cccgtttaga ggccccaagg
     7141 ggttatgcta tcaatcgttg cgttacacac acaaaaaacc aacacacatc catcttcgat
     7201 ggatagcgat tttattatct aactgctgat cgagtgtagc cagatctagt aatcaattac
     7261 ggggtcatta gttcatagcc catatatgga gttccg
//
