LOCUS       sequence-307272-        6824 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 6824)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6824)
  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-m4133715
FEATURES             Location/Qualifiers
     source          1..6824
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        86..389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        391..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     544..564
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      662..671
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1334..1354
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1355..1396
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    1449..1570
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1460..1479)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1514..1533
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1685..1731
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1761..1897
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1942..2013
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2239..2827)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2319..2338)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3327..4536
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4373..4392
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4540..5139
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4540..4559)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5036..5056
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5166..5214
                     /note="synthetic polyadenylation signal"
     CDS             join(5480..5548,5549..6340)
                     /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(5698..5717)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6353..6489
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6641..6712
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6717..6763)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    join(6781..6824,1..5)
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 ttattatcta actgctgatc gagtgtagcc agatctagta atcaattacg gggtcattag
       61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      121 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      181 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      361 tctacgtatt agtcatcgct attaccatgc tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
      601 tgaaccgtca gatcagatct ttgtcgatcc taccatccac tcgacacacc cgccagcggc
      661 cgccaccatg gcagattcca acggtactat taccgttgaa gagcttaaaa agctccttga
      721 acaatggaac ctagtaatag gtttcctatt ccttacatgg atttgtcttc tacaatttgc
      781 ctatgccaac aggaataggt ttttgtatat aattaagtta attttcctct ggctgttatg
      841 gccagtaact ttagcttgtt ttgtgcttgc tgctgtttac agaataaatt ggatcaccgg
      901 tggaattgct atcgcaatgg cttgtcttgt aggcttgatg tggctcagct acttcattgc
      961 ttctttcaga ctgtttgcgc gtacgcgttc catgtggtca ttcaatccag aaactaacat
     1021 tcttctcaac gtgccactcc atggcactat tctgaccaga ccgcttctag aaagtgaact
     1081 cgtaatcgga gctgtgatcc ttcgtggaca tcttcgtatt gctggacacc atctaggacg
     1141 ctgtgacatc aaggacctgc ctaaagaaat cactgttgct acatcacgaa cgctttctta
     1201 ttacaaattg ggagcttcgc agcgtgtagc aggtgactca ggttttgctg catacagtcg
     1261 ctacaggatt ggcaactata aattaaacac agaccattcc agtagcagtg acaatattgc
     1321 tttgcttgta caggaaaacc tttactttca gtcggggaaa cctataccca atccacttct
     1381 cggcttagat tctacctaag ctagcttgac tgactgagat acagcgtacc ttcagctcac
     1441 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     1501 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     1561 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     1621 ggaggttttt taaagcaagt aaaacctcta caaatgtggt attggcccat ctctatcggt
     1681 atcgtagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg
     1741 ggccggattg ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
     1801 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
     1861 tccatacgtg tcctccttac cagaaattta tccttaaggt cgtcagctat cctgcaggcg
     1921 atctctcgat ttcgatcaag acattccttt aatggtcttt tctggacacc actaggggtc
     1981 agaagtagtt catcaaactt tcttccctcc ctaatctcat tggttacctt gggctatcga
     2041 aacttaatta accagtcaag tcagctactt ggcgagatcg acttgtctgg gtttcgacta
     2101 cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc gttctccgat
     2161 tacgagtttc atttaaatca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2221 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2281 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2341 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2401 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2461 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2521 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2581 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2641 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     2701 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2761 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2821 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2881 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2941 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3001 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3061 tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc
     3121 gatccatctt gcaggctacc tctcgaacga actatcgcaa gtctcttggc cggccttgcg
     3181 ccttggctat tgcttggcag cgcctatcgc caggtattac tccaatcccg aatatccgag
     3241 atcgggatca cccgagagaa gttcaaccta catcctcaat cccgatctat ccgagatccg
     3301 aggaatatcg aaatcggggc gcgcctggcc tccgcgccgg gttttggcgc ctcccgcggg
     3361 cgcccccctc gtcacggcga gcgctgccac gtcagacgaa gggcgcagga gcgtcctgat
     3421 ccttccgccc ggacgctcag gacagcggcc cgctgctcat aagactcggc cttagaaccc
     3481 cagtatcagc agaaggacat tttaggacgg gacttgggtg actctagggc actggttttc
     3541 tttccagaga gcggaacagg cgaggaaaag tagtcccttc tcggcgattc tgcggaggga
     3601 tctccgtggg gcggtgaacg ccgatgatta tataaggacg cgccgggtgt ggcacagcta
     3661 gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac
     3721 ttggtgagta gcgggctgct gggctggccg gggctttcgt ggccgccggg ccgctcggtg
     3781 ggacggaagc gtgtggagag accgccaagg gctgtagtct gggtccgcga gcaaggttgc
     3841 cctgaactgg gggttggggg gagcgcagca aaatggcggc tgttcccgag tcttgaatgg
     3901 aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg
     3961 caagaaccca aggtcttgag cccttcgcta atgcgggaaa gctcttattc gggtgagatg
     4021 ggctgggcac catctgggga ccctgacgtg aagtttgtca ctgactggag aactcggttt
     4081 gtcgtctgtt gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt
     4141 gggagcgcgc gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta taatgcaggg
     4201 tggggccacc tgccggtagg tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg
     4261 ggcctagggt aggctctcct gaatcgacag gcgccggacc tctggtgagg ggagggataa
     4321 gtgaggcgtc agtttctttg gtcggtttta tgtacctatc ttcttaagta gctgaagctc
     4381 cggttttgaa ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct
     4441 tttgaaatgt aatcatttgg gtcaatatgt aattttcagt gttagacttg taaattgtcc
     4501 gctaaattct ggccgttttt ggcttttttg ttagacaaca tgaccgagta caagcccacg
     4561 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
     4621 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
     4681 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     4741 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     4801 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     4861 cagatggagg gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     4921 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     4981 gtggaggccg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     5041 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     5101 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctagg ctagtatgta agcctagtct
     5161 tagataataa aatcgctatc catcgaagat ggatgtgtgt tggttttttg tgtgtgtaac
     5221 gctaggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga cgatccatat
     5281 cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc caatcgtcag
     5341 atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta gatattgata
     5401 gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc aagagacagg
     5461 atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta ttcccttttt
     5521 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     5581 tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca gcggtaagat
     5641 ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta aagttctgct
     5701 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     5761 ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc ttacggatgg
     5821 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     5881 cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5941 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     6001 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg
     6061 cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg cggataaagt
     6121 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     6181 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     6241 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     6301 gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag gtgcattggc
     6361 gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca
     6421 gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt
     6481 tatccttaag atcccgaatc gtttaaactc gactctggct ctatcgaatc tccgtcgttt
     6541 cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct
     6601 atcgtcagct tacctttttg gcagcgatcg cggctcccga catcttggac cattagctcc
     6661 acaggtatct tcttccctct agtggtcata acagcagctt cagctacctc tcaattcaaa
     6721 aaacccctca agacccgttt agaggcccca aggggttatg ctatcaatcg ttgcgttaca
     6781 cacacaaaaa accaacacac atccatcttc gatggatagc gatt
//
