LOCUS       sequence-295593-        7610 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 7610)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7610)
  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-m4134516
FEATURES             Location/Qualifiers
     source          1..7610
                     /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             1040..1057
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1150..1271
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1161..1180)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1215..1234
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1386..1432
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1462..1598
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1643..1714
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1940..2528)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2020..2039)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3028..4237
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4074..4093
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4241..5269
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5296..5344
                     /note="synthetic polyadenylation signal"
     CDS             join(5610..5678,5679..6470)
                     /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(5828..5847)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6483..6619
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6764..6835
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6840..6886)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6904..6952
                     /note="synthetic polyadenylation signal"
     enhancer        7033..7336
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7338..7540
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7491..7511
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(7609..7610,1..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 gatgagtgac aaaatcattc atctaaccga
      721 tgactccttt gatacagacg tgttgaaggc cgatggtgcg atactggtag atttctgggc
      781 tgaatggtgt ggcccgtgca agatgatcgc acccattctg gacgagatcg ccgacgagta
      841 tcaaggaaaa ttaactgtcg ctaagcttaa tatagatcag aacccaggga cggcacctaa
      901 atacggcatc aggggaatac ctaccctcct gctctttaag aatggggaag ttgccgcgac
      961 aaaagtggga gccctgtcta agggccagct aaaagagttc ttggacgcta acctggcagg
     1021 ttcaggctcg gggcacatgc accatcacca tcaccatagc agcgggctcg tgccacgcgg
     1081 ctaaggatcc gcaggcctct gctagcttga ctgactgaga tacagcgtac cttcagctca
     1141 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     1201 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     1261 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     1321 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tattggccca tctctatcgg
     1381 tatcgtagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgtgcccctc
     1441 gggccggatt gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc
     1501 gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac
     1561 ttccatacgt gtcctcctta ccagaaattt atccttaagg tcgtcagcta tcctgcaggc
     1621 gatctctcga tttcgatcaa gacattcctt taatggtctt ttctggacac cactaggggt
     1681 cagaagtagt tcatcaaact ttcttccctc cctaatctca ttggttacct tgggctatcg
     1741 aaacttaatt aaccagtcaa gtcagctact tggcgagatc gacttgtctg ggtttcgact
     1801 acgctcagaa ttgcgtcagt caagttcgat ctggtccttg ctattgcacc cgttctccga
     1861 ttacgagttt catttaaatc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     1921 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     1981 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2041 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2101 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2161 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2221 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2281 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2341 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     2401 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2461 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     2521 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     2581 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     2641 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     2701 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     2761 ttgcatttaa atttccgaac tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc
     2821 cgatccatct tgcaggctac ctctcgaacg aactatcgca agtctcttgg ccggccttgc
     2881 gccttggcta ttgcttggca gcgcctatcg ccaggtatta ctccaatccc gaatatccga
     2941 gatcgggatc acccgagaga agttcaacct acatcctcaa tcccgatcta tccgagatcc
     3001 gaggaatatc gaaatcgggg cgcgcctggc ctccgcgccg ggttttggcg cctcccgcgg
     3061 gcgcccccct cgtcacggcg agcgctgcca cgtcagacga agggcgcagg agcgtcctga
     3121 tccttccgcc cggacgctca ggacagcggc ccgctgctca taagactcgg ccttagaacc
     3181 ccagtatcag cagaaggaca ttttaggacg ggacttgggt gactctaggg cactggtttt
     3241 ctttccagag agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg
     3301 atctccgtgg ggcggtgaac gccgatgatt atataaggac gcgccgggtg tggcacagct
     3361 agttccgtcg cagccgggat ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca
     3421 cttggtgagt agcgggctgc tgggctggcc ggggctttcg tggccgccgg gccgctcggt
     3481 gggacggaag cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg
     3541 ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg
     3601 gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg
     3661 gcaagaaccc aaggtcttga gcccttcgct aatgcgggaa agctcttatt cgggtgagat
     3721 gggctgggca ccatctgggg accctgacgt gaagtttgtc actgactgga gaactcggtt
     3781 tgtcgtctgt tgcgggggcg gcagttatgg cggtgccgtt gggcagtgca cccgtacctt
     3841 tgggagcgcg cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg
     3901 gtggggccac ctgccggtag gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc
     3961 gggcctaggg taggctctcc tgaatcgaca ggcgccggac ctctggtgag gggagggata
     4021 agtgaggcgt cagtttcttt ggtcggtttt atgtacctat cttcttaagt agctgaagct
     4081 ccggttttga actatgcgct cggggttggc gagtgtgttt tgtgaagttt tttaggcacc
     4141 ttttgaaatg taatcatttg ggtcaatatg taattttcag tgttagactt gtaaattgtc
     4201 cgctaaattc tggccgtttt tggctttttt gttagacaac atgggtaaaa agcctgaact
     4261 caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat
     4321 gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata
     4381 tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca
     4441 ctttgcatcc gccgcgctcc cgattccgga agtgcttgac attggggagt tcagcgagag
     4501 cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac
     4561 cgaactgccc gctgttctgc agccggtcgc ggaggcaatg gatgccatcg ctgccgccga
     4621 tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac
     4681 atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat
     4741 ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga
     4801 ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac
     4861 ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca
     4921 atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac
     4981 gcgctacttc gagcggaggc atccggagct tgcaggatcg ccccggctcc gggcgtatat
     5041 gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc
     5101 agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg
     5161 tacacaaatc gcccgcagaa gcgccgccgt ctggaccgat ggctgtgtag aagtactcgc
     5221 cgatagtgga aaccgacgcc ccagcactcg tccgagggca aaggaataag ctagtatgta
     5281 agcctagtct tagataataa aatcgctatc catcgaagat ggatgtgtgt tggttttttg
     5341 tgtgtgtaac gctaggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga
     5401 cgatccatat cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc
     5461 caatcgtcag atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta
     5521 gatattgata gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc
     5581 aagagacagg atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta
     5641 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     5701 taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca
     5761 gcggtaagat ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta
     5821 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     5881 gccgcataca ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc
     5941 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     6001 ctgcggccaa cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc
     6061 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     6121 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac
     6181 tattaactgg cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg
     6241 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     6301 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     6361 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     6421 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag
     6481 gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat
     6541 gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt
     6601 accagaaatt tatccttaag atcgtttaaa ctcgactctg gctctatcga atctccgtcg
     6661 tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgggcatcg aatctcgtca
     6721 gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg gaccattagc
     6781 tccacaggta tcttcttccc tctagtggtc ataacagcag cttcagctac ctctcaattc
     6841 aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt
     6901 acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta ttatctaact
     6961 gctgatcgag tgtagccaga tctagtaatc aattacgggg tcattagttc atagcccata
     7021 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     7081 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     7141 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     7201 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     7261 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     7321 catcgctatt accatgctga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     7381 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     7441 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     7501 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
     7561 cagatctttg tcgatcctac catccactcg acacacccgc cagcggccgc
//
