LOCUS       sequence-319546-        7269 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 7269)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7269)
  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-m4133594
FEATURES             Location/Qualifiers
     source          1..7269
                     /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..1311
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    1380..1501
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1391..1410)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1445..1464
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1616..1662
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1692..1828
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1873..1944
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2170..2758)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2250..2269)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3258..4467
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4304..4323
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4471..5499
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5526..5574
                     /note="synthetic polyadenylation signal"
     CDS             join(5840..5908,5909..6700)
                     /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(6058..6077)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6713..6849
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7001..7072
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(7077..7123)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    7141..7189
                     /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 tttcagtcgg ggaaacctat acccaatcca cttctcggct tagattctac ctaaggatcc
     1321 gcaggcctct gctagcttga ctgactgaga tacagcgtac cttcagctca cagacatgat
     1381 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     1441 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     1501 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt
     1561 ttaaagcaag taaaacctct acaaatgtgg tattggccca tctctatcgg tatcgtagca
     1621 taaccccttg gggcctctaa acgggtcttg aggggttttt tgtgcccctc gggccggatt
     1681 gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
     1741 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
     1801 gtcctcctta ccagaaattt atccttaagg tcgtcagcta tcctgcaggc gatctctcga
     1861 tttcgatcaa gacattcctt taatggtctt ttctggacac cactaggggt cagaagtagt
     1921 tcatcaaact ttcttccctc cctaatctca ttggttacct tgggctatcg aaacttaatt
     1981 aaccagtcaa gtcagctact tggcgagatc gacttgtctg ggtttcgact acgctcagaa
     2041 ttgcgtcagt caagttcgat ctggtccttg ctattgcacc cgttctccga ttacgagttt
     2101 catttaaatc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2161 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2221 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2281 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2341 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2401 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2461 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2521 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2581 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     2641 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     2701 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2761 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     2821 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     2881 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     2941 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcatttaa
     3001 atttccgaac tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc cgatccatct
     3061 tgcaggctac ctctcgaacg aactatcgca agtctcttgg ccggccttgc gccttggcta
     3121 ttgcttggca gcgcctatcg ccaggtatta ctccaatccc gaatatccga gatcgggatc
     3181 acccgagaga agttcaacct acatcctcaa tcccgatcta tccgagatcc gaggaatatc
     3241 gaaatcgggg cgcgcctggc ctccgcgccg ggttttggcg cctcccgcgg gcgcccccct
     3301 cgtcacggcg agcgctgcca cgtcagacga agggcgcagg agcgtcctga tccttccgcc
     3361 cggacgctca ggacagcggc ccgctgctca taagactcgg ccttagaacc ccagtatcag
     3421 cagaaggaca ttttaggacg ggacttgggt gactctaggg cactggtttt ctttccagag
     3481 agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt ctgcggaggg atctccgtgg
     3541 ggcggtgaac gccgatgatt atataaggac gcgccgggtg tggcacagct agttccgtcg
     3601 cagccgggat ttgggtcgcg gttcttgttt gtggatcgct gtgatcgtca cttggtgagt
     3661 agcgggctgc tgggctggcc ggggctttcg tggccgccgg gccgctcggt gggacggaag
     3721 cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg agcaaggttg ccctgaactg
     3781 ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga gtcttgaatg gaagacgctt
     3841 gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc atggtgggcg gcaagaaccc
     3901 aaggtcttga gcccttcgct aatgcgggaa agctcttatt cgggtgagat gggctgggca
     3961 ccatctgggg accctgacgt gaagtttgtc actgactgga gaactcggtt tgtcgtctgt
     4021 tgcgggggcg gcagttatgg cggtgccgtt gggcagtgca cccgtacctt tgggagcgcg
     4081 cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt ataatgcagg gtggggccac
     4141 ctgccggtag gtgtgcggta ggcttttctc cgtcgcagga cgcagggttc gggcctaggg
     4201 taggctctcc tgaatcgaca ggcgccggac ctctggtgag gggagggata agtgaggcgt
     4261 cagtttcttt ggtcggtttt atgtacctat cttcttaagt agctgaagct ccggttttga
     4321 actatgcgct cggggttggc gagtgtgttt tgtgaagttt tttaggcacc ttttgaaatg
     4381 taatcatttg ggtcaatatg taattttcag tgttagactt gtaaattgtc cgctaaattc
     4441 tggccgtttt tggctttttt gttagacaac atgggtaaaa agcctgaact caccgcgacg
     4501 tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg
     4561 gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg
     4621 gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcc
     4681 gccgcgctcc cgattccgga agtgcttgac attggggagt tcagcgagag cctgacctat
     4741 tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc
     4801 gctgttctgc agccggtcgc ggaggcaatg gatgccatcg ctgccgccga tcttagccag
     4861 acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat
     4921 ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc
     4981 gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc
     5041 gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc
     5101 cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc
     5161 gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc
     5221 gagcggaggc atccggagct tgcaggatcg ccccggctcc gggcgtatat gctccgcatt
     5281 ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg
     5341 cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc
     5401 gcccgcagaa gcgccgccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga
     5461 aaccgacgcc ccagcactcg tccgagggca aaggaataag ctagtatgta agcctagtct
     5521 tagataataa aatcgctatc catcgaagat ggatgtgtgt tggttttttg tgtgtgtaac
     5581 gctaggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga cgatccatat
     5641 cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc caatcgtcag
     5701 atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta gatattgata
     5761 gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc aagagacagg
     5821 atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta ttcccttttt
     5881 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     5941 tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca gcggtaagat
     6001 ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta aagttctgct
     6061 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     6121 ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc ttacggatgg
     6181 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     6241 cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     6301 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     6361 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg
     6421 cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg cggataaagt
     6481 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     6541 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     6601 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     6661 gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag gtgcattggc
     6721 gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca
     6781 gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt
     6841 tatccttaag atcccgaatc gtttaaactc gactctggct ctatcgaatc tccgtcgttt
     6901 cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct
     6961 atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac cattagctcc
     7021 acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc tcaattcaaa
     7081 aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg ttgcgttaca
     7141 cacacaaaaa accaacacac atccatcttc gatggatagc gattttatta tctaactgct
     7201 gatcgagtgt agccagatct agtaatcaat tacggggtca ttagttcata gcccatatat
     7261 ggagttccg
//
