LOCUS       sequence-295602-        7238 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 7238)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7238)
  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-m4134338
FEATURES             Location/Qualifiers
     source          1..7238
                     /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..709
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    778..899
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(789..808)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     843..862
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1014..1060
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1090..1226
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1271..1342
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1568..2156)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1648..1667)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2656..3865
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3702..3721
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3869..4897
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    4924..4972
                     /note="synthetic polyadenylation signal"
     CDS             join(5238..5306,5307..6098)
                     /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(5456..5475)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6111..6247
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6392..6463
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6468..6514)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6532..6580
                     /note="synthetic polyadenylation signal"
     enhancer        6661..6964
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6966..7168
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7119..7139
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(7237..7238,1..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 gcaccatcac catcaccatt aaggatccgc
      721 aggcctctgc tagcttgact gactgagata cagcgtacct tcagctcaca gacatgataa
      781 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
      841 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta
      901 acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt
      961 aaagcaagta aaacctctac aaatgtggta ttggcccatc tctatcggta tcgtagcata
     1021 accccttggg gcctctaaac gggtcttgag gggttttttg tgcccctcgg gccggattgc
     1081 tatctaccgg cattggcgca gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc
     1141 cacatatgcc agattcagca acggatacgg cttccccaac ttgcccactt ccatacgtgt
     1201 cctccttacc agaaatttat ccttaaggtc gtcagctatc ctgcaggcga tctctcgatt
     1261 tcgatcaaga cattccttta atggtctttt ctggacacca ctaggggtca gaagtagttc
     1321 atcaaacttt cttccctccc taatctcatt ggttaccttg ggctatcgaa acttaattaa
     1381 ccagtcaagt cagctacttg gcgagatcga cttgtctggg tttcgactac gctcagaatt
     1441 gcgtcagtca agttcgatct ggtccttgct attgcacccg ttctccgatt acgagtttca
     1501 tttaaatcat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     1561 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     1621 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     1681 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     1741 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     1801 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     1861 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     1921 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     1981 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     2041 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     2101 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     2161 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     2221 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     2281 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     2341 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcatttaaat
     2401 ttccgaactc tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg atccatcttg
     2461 caggctacct ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc cttggctatt
     2521 gcttggcagc gcctatcgcc aggtattact ccaatcccga atatccgaga tcgggatcac
     2581 ccgagagaag ttcaacctac atcctcaatc ccgatctatc cgagatccga ggaatatcga
     2641 aatcggggcg cgcctggcct ccgcgccggg ttttggcgcc tcccgcgggc gcccccctcg
     2701 tcacggcgag cgctgccacg tcagacgaag ggcgcaggag cgtcctgatc cttccgcccg
     2761 gacgctcagg acagcggccc gctgctcata agactcggcc ttagaacccc agtatcagca
     2821 gaaggacatt ttaggacggg acttgggtga ctctagggca ctggttttct ttccagagag
     2881 cggaacaggc gaggaaaagt agtcccttct cggcgattct gcggagggat ctccgtgggg
     2941 cggtgaacgc cgatgattat ataaggacgc gccgggtgtg gcacagctag ttccgtcgca
     3001 gccgggattt gggtcgcggt tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag
     3061 cgggctgctg ggctggccgg ggctttcgtg gccgccgggc cgctcggtgg gacggaagcg
     3121 tgtggagaga ccgccaaggg ctgtagtctg ggtccgcgag caaggttgcc ctgaactggg
     3181 ggttgggggg agcgcagcaa aatggcggct gttcccgagt cttgaatgga agacgcttgt
     3241 gaggcgggct gtgaggtcgt tgaaacaagg tggggggcat ggtgggcggc aagaacccaa
     3301 ggtcttgagc ccttcgctaa tgcgggaaag ctcttattcg ggtgagatgg gctgggcacc
     3361 atctggggac cctgacgtga agtttgtcac tgactggaga actcggtttg tcgtctgttg
     3421 cgggggcggc agttatggcg gtgccgttgg gcagtgcacc cgtacctttg ggagcgcgcg
     3481 ccctcgtcgt gtcgtgacgt cacccgttct gttggcttat aatgcagggt ggggccacct
     3541 gccggtaggt gtgcggtagg cttttctccg tcgcaggacg cagggttcgg gcctagggta
     3601 ggctctcctg aatcgacagg cgccggacct ctggtgaggg gagggataag tgaggcgtca
     3661 gtttctttgg tcggttttat gtacctatct tcttaagtag ctgaagctcc ggttttgaac
     3721 tatgcgctcg gggttggcga gtgtgttttg tgaagttttt taggcacctt ttgaaatgta
     3781 atcatttggg tcaatatgta attttcagtg ttagacttgt aaattgtccg ctaaattctg
     3841 gccgtttttg gcttttttgt tagacaacat gggtaaaaag cctgaactca ccgcgacgtc
     3901 tgtcgagaag tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga
     3961 gggcgaagaa tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt
     4021 aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatccgc
     4081 cgcgctcccg attccggaag tgcttgacat tggggagttc agcgagagcc tgacctattg
     4141 catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc
     4201 tgttctgcag ccggtcgcgg aggcaatgga tgccatcgct gccgccgatc ttagccagac
     4261 gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt
     4321 catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt
     4381 cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga
     4441 agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg
     4501 cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc
     4561 caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga
     4621 gcggaggcat ccggagcttg caggatcgcc ccggctccgg gcgtatatgc tccgcattgg
     4681 tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca
     4741 gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc
     4801 ccgcagaagc gccgccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa
     4861 ccgacgcccc agcactcgtc cgagggcaaa ggaataagct agtatgtaag cctagtctta
     4921 gataataaaa tcgctatcca tcgaagatgg atgtgtgttg gttttttgtg tgtgtaacgc
     4981 taggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg
     5041 ttgcttggca gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat
     5101 attgtactca agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt
     5161 ctgatcggtc aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat
     5221 cagcaggagg ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     5281 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5341 aagatcagtt gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5401 ttgagagttt tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat
     5461 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5521 attctcagaa tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca
     5581 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5641 tacttctgac aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5701 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5761 agcgtgacac cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg
     5821 aactacttac tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg
     5881 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5941 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     6001 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     6061 tcgctgagat aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc
     6121 agaaaaaaat gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc
     6181 aacggatacg gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta
     6241 tccttaagat cgtttaaact cgactctggc tctatcgaat ctccgtcgtt tcgagcttac
     6301 gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa tctcgtcagc tatcgtcagc
     6361 ttaccttttt ggcagcgatc gcggctcccg acatcttgga ccattagctc cacaggtatc
     6421 ttcttccctc tagtggtcat aacagcagct tcagctacct ctcaattcaa aaaacccctc
     6481 aagacccgtt tagaggcccc aaggggttat gctatcaatc gttgcgttac acacacaaaa
     6541 aaccaacaca catccatctt cgatggatag cgattttatt atctaactgc tgatcgagtg
     6601 tagccagatc tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     6661 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     6721 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     6781 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     6841 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     6901 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     6961 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
     7021 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     7081 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
     7141 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatca gatctttgtc
     7201 gatcctacca tccactcgac acacccgcca gcggccgc
//
