LOCUS       sequence-313509-        7254 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 7254)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7254)
  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-m4133607
FEATURES             Location/Qualifiers
     source          1..7254
                     /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             1270..1296
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1365..1486
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1376..1395)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1430..1449
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1601..1647
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1677..1813
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1858..1929
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2155..2743)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2235..2254)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3243..4452
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4289..4308
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4456..5484
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5511..5559
                     /note="synthetic polyadenylation signal"
     CDS             join(5825..5893,5894..6685)
                     /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(6043..6062)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6698..6834
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6986..7057
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7062..7108)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7126..7174
                     /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 tttcagtcgt acccctatga cgtaccagat tatgcgtaag gatccgcagg cctctgctag
     1321 cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat acattgatga
     1381 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     1441 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     1501 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     1561 cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc ccttggggcc
     1621 tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat ctaccggcat
     1681 tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga
     1741 ttcagcaacg gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga
     1801 aatttatcct taaggtcgtc agctatcctg caggcgatct ctcgatttcg atcaagacat
     1861 tcctttaatg gtcttttctg gacaccacta ggggtcagaa gtagttcatc aaactttctt
     1921 ccctccctaa tctcattggt taccttgggc tatcgaaact taattaacca gtcaagtcag
     1981 ctacttggcg agatcgactt gtctgggttt cgactacgct cagaattgcg tcagtcaagt
     2041 tcgatctggt ccttgctatt gcacccgttc tccgattacg agtttcattt aaatcatgtg
     2101 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2161 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2221 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2281 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2341 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2401 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2461 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2521 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2581 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2641 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2701 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2761 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2821 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     2881 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     2941 tatctcagcg atctgtctat ttcgttcatc catagttgca tttaaatttc cgaactctcc
     3001 aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc catcttgcag gctacctctc
     3061 gaacgaacta tcgcaagtct cttggccggc cttgcgcctt ggctattgct tggcagcgcc
     3121 tatcgccagg tattactcca atcccgaata tccgagatcg ggatcacccg agagaagttc
     3181 aacctacatc ctcaatcccg atctatccga gatccgagga atatcgaaat cggggcgcgc
     3241 ctggcctccg cgccgggttt tggcgcctcc cgcgggcgcc cccctcgtca cggcgagcgc
     3301 tgccacgtca gacgaagggc gcaggagcgt cctgatcctt ccgcccggac gctcaggaca
     3361 gcggcccgct gctcataaga ctcggcctta gaaccccagt atcagcagaa ggacatttta
     3421 ggacgggact tgggtgactc tagggcactg gttttctttc cagagagcgg aacaggcgag
     3481 gaaaagtagt cccttctcgg cgattctgcg gagggatctc cgtggggcgg tgaacgccga
     3541 tgattatata aggacgcgcc gggtgtggca cagctagttc cgtcgcagcc gggatttggg
     3601 tcgcggttct tgtttgtgga tcgctgtgat cgtcacttgg tgagtagcgg gctgctgggc
     3661 tggccggggc tttcgtggcc gccgggccgc tcggtgggac ggaagcgtgt ggagagaccg
     3721 ccaagggctg tagtctgggt ccgcgagcaa ggttgccctg aactgggggt tggggggagc
     3781 gcagcaaaat ggcggctgtt cccgagtctt gaatggaaga cgcttgtgag gcgggctgtg
     3841 aggtcgttga aacaaggtgg ggggcatggt gggcggcaag aacccaaggt cttgagccct
     3901 tcgctaatgc gggaaagctc ttattcgggt gagatgggct gggcaccatc tggggaccct
     3961 gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg gggcggcagt
     4021 tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc
     4081 gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccacctgcc ggtaggtgtg
     4141 cggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc tctcctgaat
     4201 cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt tctttggtcg
     4261 gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat gcgctcgggg
     4321 ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca
     4381 atatgtaatt ttcagtgtta gacttgtaaa ttgtccgcta aattctggcc gtttttggct
     4441 tttttgttag acaacatggg taaaaagcct gaactcaccg cgacgtctgt cgagaagttt
     4501 ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg cgaagaatct
     4561 cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc
     4621 gatggtttct acaaagatcg ttatgtttat cggcactttg catccgccgc gctcccgatt
     4681 ccggaagtgc ttgacattgg ggagttcagc gagagcctga cctattgcat ctcccgccgt
     4741 gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt tctgcagccg
     4801 gtcgcggagg caatggatgc catcgctgcc gccgatctta gccagacgag cgggttcggc
     4861 ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat atgcgcgatt
     4921 gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc
     4981 gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt ccggcacctc
     5041 gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat aacagcggtc
     5101 attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa catcttcttc
     5161 tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg gaggcatccg
     5221 gagcttgcag gatcgccccg gctccgggcg tatatgctcc gcattggtct tgaccaactc
     5281 tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg tcgatgcgac
     5341 gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg cagaagcgcc
     5401 gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg acgccccagc
     5461 actcgtccga gggcaaagga ataagctagt atgtaagcct agtcttagat aataaaatcg
     5521 ctatccatcg aagatggatg tgtgttggtt ttttgtgtgt gtaacgctag gcgcgcctgg
     5581 tgtaccgaga acgatcctct cagtgcgagt ctcgacgatc catatcgttg cttggcagtc
     5641 agccagtcgg aatccagctt gggacccagg aagtccaatc gtcagatatt gtactcaagc
     5701 ctggtcacgg cagcgtaccg atctgtttaa acctagatat tgatagtctg atcggtcaac
     5761 gtataatcga gtcctagctt ttgcaaacat ctatcaagag acaggatcag caggaggctt
     5821 tcgcatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5881 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5941 tgcgcgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     6001 ccccgaagaa cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     6061 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     6121 cttggttgag tattcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     6181 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     6241 gattggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     6301 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     6361 gatgcctgta gcaatggcaa caaccttgcg taaactatta actggcgaac tacttactct
     6421 agcttcccgg caacagttga tagactggat ggaggcggat aaagttgcag gaccacttct
     6481 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     6541 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     6601 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     6661 tgcctcactg attaagcatt ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc
     6721 tgatgcgacg ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct
     6781 tccccaactt gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaagatccc
     6841 gaatcgttta aactcgactc tggctctatc gaatctccgt cgtttcgagc ttacgcgaac
     6901 agccgtggcg ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt cagcttacct
     6961 ttttggcagc gatcgcggct cccgacatct tggaccatta gctccacagg tatcttcttc
     7021 cctctagtgg tcataacagc agcttcagct acctctcaat tcaaaaaacc cctcaagacc
     7081 cgtttagagg ccccaagggg ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa
     7141 cacacatcca tcttcgatgg atagcgattt tattatctaa ctgctgatcg agtgtagcca
     7201 gatctagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccg
//
