LOCUS       sequence-307296-        7272 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 7272)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 7272)
  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-m4133271
FEATURES             Location/Qualifiers
     source          1..7272
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    53..124
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(129..175)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    193..241
                     /note="synthetic polyadenylation signal"
     enhancer        322..625
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        627..829
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     780..800
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      898..907
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1570..1590
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1591..1635
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    1704..1825
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1715..1734)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1769..1788
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1940..1986
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2016..2152
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2197..2268
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2494..3082)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2574..2593)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3582..4791
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4628..4647
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4795..5823
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     polyA_signal    5850..5898
                     /note="synthetic polyadenylation signal"
     CDS             join(6164..6232,6233..7024)
                     /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(6382..6401)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      7037..7173
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 atctcgtcag ctatcgtcag cttacctttt tggcagcgat cgcggctccc gacatcttgg
       61 accattagct ccacaggtat cttcttccct ctagtggtca taacagcagc ttcagctacc
      121 tctcaattca aaaaacccct caagacccgt ttagaggccc caaggggtta tgctatcaat
      181 cgttgcgtta cacacacaaa aaaccaacac acatccatct tcgatggata gcgattttat
      241 tatctaactg ctgatcgagt gtagccagat ctagtaatca attacggggt cattagttca
      301 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      361 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      421 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      481 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      541 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      601 cgtattagtc atcgctatta ccatgctgat gcggttttgg cagtacatca atgggcgtgg
      661 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      721 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      781 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
      841 ccgtcagatc agatctttgt cgatcctacc atccactcga cacacccgcc agcggccgcc
      901 accatggcag attccaacgg tactattacc gttgaagagc ttaaaaagct ccttgaacaa
      961 tggaacctag taataggttt cctattcctt acatggattt gtcttctaca atttgcctat
     1021 gccaacagga ataggttttt gtatataatt aagttaattt tcctctggct gttatggcca
     1081 gtaactttag cttgttttgt gcttgctgct gtttacagaa taaattggat caccggtgga
     1141 attgctatcg caatggcttg tcttgtaggc ttgatgtggc tcagctactt cattgcttct
     1201 ttcagactgt ttgcgcgtac gcgttccatg tggtcattca atccagaaac taacattctt
     1261 ctcaacgtgc cactccatgg cactattctg accagaccgc ttctagaaag tgaactcgta
     1321 atcggagctg tgatccttcg tggacatctt cgtattgctg gacaccatct aggacgctgt
     1381 gacatcaagg acctgcctaa agaaatcact gttgctacat cacgaacgct ttcttattac
     1441 aaattgggag cttcgcagcg tgtagcaggt gactcaggtt ttgctgcata cagtcgctac
     1501 aggattggca actataaatt aaacacagac cattccagta gcagtgacaa tattgctttg
     1561 cttgtacagg aaaaccttta ctttcagtcg aaagaaaccg cggcagctaa gttcgagagg
     1621 cagcacatgg acagttaagg atccgcaggc ctctgctagc ttgactgact gagatacagc
     1681 gtaccttcag ctcacagaca tgataagata cattgatgag tttggacaaa ccacaactag
     1741 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     1801 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     1861 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtattgg
     1921 cccatctcta tcggtatcgt agcataaccc cttggggcct ctaaacgggt cttgaggggt
     1981 tttttgtgcc cctcgggccg gattgctatc taccggcatt ggcgcagaaa aaaatgcctg
     2041 atgcgacgct gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc
     2101 cccaacttgc ccacttccat acgtgtcctc cttaccagaa atttatcctt aaggtcgtca
     2161 gctatcctgc aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg
     2221 acaccactag gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt
     2281 accttgggct atcgaaactt aattaaccag tcaagtcagc tacttggcga gatcgacttg
     2341 tctgggtttc gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg
     2401 cacccgttct ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc
     2461 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2521 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2581 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2641 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2701 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2761 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2821 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2881 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     2941 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3001 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3061 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3121 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3181 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3241 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3301 tcgttcatcc atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct
     3361 tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc
     3421 ttggccggcc ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa
     3481 tcccgaatat ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga
     3541 tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc tggcctccgc gccgggtttt
     3601 ggcgcctccc gcgggcgccc ccctcgtcac ggcgagcgct gccacgtcag acgaagggcg
     3661 caggagcgtc ctgatccttc cgcccggacg ctcaggacag cggcccgctg ctcataagac
     3721 tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt gggtgactct
     3781 agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc ccttctcggc
     3841 gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatataa ggacgcgccg
     3901 ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt gtttgtggat
     3961 cgctgtgatc gtcacttggt gagtagcggg ctgctgggct ggccggggct ttcgtggccg
     4021 ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc caagggctgt agtctgggtc
     4081 cgcgagcaag gttgccctga actgggggtt ggggggagcg cagcaaaatg gcggctgttc
     4141 ccgagtcttg aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa acaaggtggg
     4201 gggcatggtg ggcggcaaga acccaaggtc ttgagccctt cgctaatgcg ggaaagctct
     4261 tattcgggtg agatgggctg ggcaccatct ggggaccctg acgtgaagtt tgtcactgac
     4321 tggagaactc ggtttgtcgt ctgttgcggg ggcggcagtt atggcggtgc cgttgggcag
     4381 tgcacccgta cctttgggag cgcgcgccct cgtcgtgtcg tgacgtcacc cgttctgttg
     4441 gcttataatg cagggtgggg ccacctgccg gtaggtgtgc ggtaggcttt tctccgtcgc
     4501 aggacgcagg gttcgggcct agggtaggct ctcctgaatc gacaggcgcc ggacctctgg
     4561 tgaggggagg gataagtgag gcgtcagttt ctttggtcgg ttttatgtac ctatcttctt
     4621 aagtagctga agctccggtt ttgaactatg cgctcggggt tggcgagtgt gttttgtgaa
     4681 gttttttagg caccttttga aatgtaatca tttgggtcaa tatgtaattt tcagtgttag
     4741 acttgtaaat tgtccgctaa attctggccg tttttggctt ttttgttaga caacatgggt
     4801 aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc
     4861 gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta
     4921 ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt
     4981 tatgtttatc ggcactttgc atccgccgcg ctcccgattc cggaagtgct tgacattggg
     5041 gagttcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa
     5101 gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc aatggatgcc
     5161 atcgctgccg ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc
     5221 ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac
     5281 tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg
     5341 atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc
     5401 aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag cgaggcgatg
     5461 ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg gttggcttgt
     5521 atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg atcgccccgg
     5581 ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt ggttgacggc
     5641 aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg atccggagcc
     5701 gggactgtcg ggcgtacaca aatcgcccgc agaagcgccg ccgtctggac cgatggctgt
     5761 gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa
     5821 taagctagta tgtaagccta gtcttagata ataaaatcgc tatccatcga agatggatgt
     5881 gtgttggttt tttgtgtgtg taacgctagg cgcgcctggt gtaccgagaa cgatcctctc
     5941 agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga atccagcttg
     6001 ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc agcgtaccga
     6061 tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag tcctagcttt
     6121 tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta ttcaacattt
     6181 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     6241 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg gttacatcga
     6301 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gctttccaat
     6361 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     6421 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt attcaccagt
     6481 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     6541 catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac cgaaggagct
     6601 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     6661 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     6721 aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc aacagttgat
     6781 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     6841 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     6901 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     6961 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     7021 gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt
     7081 atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca
     7141 tacgtgtcct ccttaccaga aatttatcct taagatcccg aatcgtttaa actcgactct
     7201 ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc
     7261 gtcgggcatc ga
//
