LOCUS       sequence-295949-        6820 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 6820)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6820)
  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-m4133733
FEATURES             Location/Qualifiers
     source          1..6820
                     /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..736
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    789..910
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(800..819)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     854..873
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1025..1071
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1101..1237
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1282..1353
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1579..2167)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1659..1678)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2667..3876
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     3713..3732
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             3880..4479
                     /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(3880..3899)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4376..4396
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4506..4554
                     /note="synthetic polyadenylation signal"
     CDS             join(4820..4888,4889..5680)
                     /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(5038..5057)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5693..5829
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5974..6045
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6050..6096)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6114..6162
                     /note="synthetic polyadenylation signal"
     enhancer        6243..6546
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6548..6750
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6701..6721
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(6819..6820,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggca gattccaacg gtactattac cgttgaagag cttaaaaagc tccttgaaca
       61 atggaaccta gtaataggtt tcctattcct tacatggatt tgtcttctac aatttgccta
      121 tgccaacagg aataggtttt tgtatataat taagttaatt ttcctctggc tgttatggcc
      181 agtaacttta gcttgttttg tgcttgctgc tgtttacaga ataaattgga tcaccggtgg
      241 aattgctatc gcaatggctt gtcttgtagg cttgatgtgg ctcagctact tcattgcttc
      301 tttcagactg tttgcgcgta cgcgttccat gtggtcattc aatccagaaa ctaacattct
      361 tctcaacgtg ccactccatg gcactattct gaccagaccg cttctagaaa gtgaactcgt
      421 aatcggagct gtgatccttc gtggacatct tcgtattgct ggacaccatc taggacgctg
      481 tgacatcaag gacctgccta aagaaatcac tgttgctaca tcacgaacgc tttcttatta
      541 caaattggga gcttcgcagc gtgtagcagg tgactcaggt tttgctgcat acagtcgcta
      601 caggattggc aactataaat taaacacaga ccattccagt agcagtgaca atattgcttt
      661 gcttgtacag gaaaaccttt actttcagtc gaaagaaacc gcggcagcta agttcgagag
      721 gcagcacatg gacagttaag ctagcttgac tgactgagat acagcgtacc ttcagctcac
      781 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
      841 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
      901 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
      961 ggaggttttt taaagcaagt aaaacctcta caaatgtggt attggcccat ctctatcggt
     1021 atcgtagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg
     1081 ggccggattg ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
     1141 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
     1201 tccatacgtg tcctccttac cagaaattta tccttaaggt cgtcagctat cctgcaggcg
     1261 atctctcgat ttcgatcaag acattccttt aatggtcttt tctggacacc actaggggtc
     1321 agaagtagtt catcaaactt tcttccctcc ctaatctcat tggttacctt gggctatcga
     1381 aacttaatta accagtcaag tcagctactt ggcgagatcg acttgtctgg gtttcgacta
     1441 cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc gttctccgat
     1501 tacgagtttc atttaaatca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1561 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1621 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1681 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1741 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1801 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1861 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1921 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1981 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     2041 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2101 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2161 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2221 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2281 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2341 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2401 tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc
     2461 gatccatctt gcaggctacc tctcgaacga actatcgcaa gtctcttggc cggccttgcg
     2521 ccttggctat tgcttggcag cgcctatcgc caggtattac tccaatcccg aatatccgag
     2581 atcgggatca cccgagagaa gttcaaccta catcctcaat cccgatctat ccgagatccg
     2641 aggaatatcg aaatcggggc gcgcctggcc tccgcgccgg gttttggcgc ctcccgcggg
     2701 cgcccccctc gtcacggcga gcgctgccac gtcagacgaa gggcgcagga gcgtcctgat
     2761 ccttccgccc ggacgctcag gacagcggcc cgctgctcat aagactcggc cttagaaccc
     2821 cagtatcagc agaaggacat tttaggacgg gacttgggtg actctagggc actggttttc
     2881 tttccagaga gcggaacagg cgaggaaaag tagtcccttc tcggcgattc tgcggaggga
     2941 tctccgtggg gcggtgaacg ccgatgatta tataaggacg cgccgggtgt ggcacagcta
     3001 gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg tggatcgctg tgatcgtcac
     3061 ttggtgagta gcgggctgct gggctggccg gggctttcgt ggccgccggg ccgctcggtg
     3121 ggacggaagc gtgtggagag accgccaagg gctgtagtct gggtccgcga gcaaggttgc
     3181 cctgaactgg gggttggggg gagcgcagca aaatggcggc tgttcccgag tcttgaatgg
     3241 aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag gtggggggca tggtgggcgg
     3301 caagaaccca aggtcttgag cccttcgcta atgcgggaaa gctcttattc gggtgagatg
     3361 ggctgggcac catctgggga ccctgacgtg aagtttgtca ctgactggag aactcggttt
     3421 gtcgtctgtt gcgggggcgg cagttatggc ggtgccgttg ggcagtgcac ccgtaccttt
     3481 gggagcgcgc gccctcgtcg tgtcgtgacg tcacccgttc tgttggctta taatgcaggg
     3541 tggggccacc tgccggtagg tgtgcggtag gcttttctcc gtcgcaggac gcagggttcg
     3601 ggcctagggt aggctctcct gaatcgacag gcgccggacc tctggtgagg ggagggataa
     3661 gtgaggcgtc agtttctttg gtcggtttta tgtacctatc ttcttaagta gctgaagctc
     3721 cggttttgaa ctatgcgctc ggggttggcg agtgtgtttt gtgaagtttt ttaggcacct
     3781 tttgaaatgt aatcatttgg gtcaatatgt aattttcagt gttagacttg taaattgtcc
     3841 gctaaattct ggccgttttt ggcttttttg ttagacaaca tgaccgagta caagcccacg
     3901 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
     3961 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
     4021 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     4081 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     4141 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     4201 cagatggagg gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     4261 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     4321 gtggaggccg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     4381 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     4441 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctagg ctagtatgta agcctagtct
     4501 tagataataa aatcgctatc catcgaagat ggatgtgtgt tggttttttg tgtgtgtaac
     4561 gctaggcgcg cctggtgtac cgagaacgat cctctcagtg cgagtctcga cgatccatat
     4621 cgttgcttgg cagtcagcca gtcggaatcc agcttgggac ccaggaagtc caatcgtcag
     4681 atattgtact caagcctggt cacggcagcg taccgatctg tttaaaccta gatattgata
     4741 gtctgatcgg tcaacgtata atcgagtcct agcttttgca aacatctatc aagagacagg
     4801 atcagcagga ggctttcgca tgagtattca acatttccgt gtcgccctta ttcccttttt
     4861 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4921 tgaagatcag ttgggtgcgc gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4981 ccttgagagt tttcgccccg aagaacgctt tccaatgatg agcactttta aagttctgct
     5041 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     5101 ctattctcag aatgacttgg ttgagtattc accagtcaca gaaaagcatc ttacggatgg
     5161 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     5221 cttacttctg acaacgattg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5281 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     5341 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc ttgcgtaaac tattaactgg
     5401 cgaactactt actctagctt cccggcaaca gttgatagac tggatggagg cggataaagt
     5461 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     5521 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5581 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5641 gatcgctgag ataggtgcct cactgattaa gcattggtaa ccgattctag gtgcattggc
     5701 gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca
     5761 gcaacggata cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt
     5821 tatccttaag atcgtttaaa ctcgactctg gctctatcga atctccgtcg tttcgagctt
     5881 acgcgaacag ccgtggcgct catttgctcg tcgggcatcg aatctcgtca gctatcgtca
     5941 gcttaccttt ttggcagcga tcgcggctcc cgacatcttg gaccattagc tccacaggta
     6001 tcttcttccc tctagtggtc ataacagcag cttcagctac ctctcaattc aaaaaacccc
     6061 tcaagacccg tttagaggcc ccaaggggtt atgctatcaa tcgttgcgtt acacacacaa
     6121 aaaaccaaca cacatccatc ttcgatggat agcgatttta ttatctaact gctgatcgag
     6181 tgtagccaga tctagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     6241 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     6301 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     6361 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     6421 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     6481 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     6541 accatgctga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
     6601 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     6661 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
     6721 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat cagatctttg
     6781 tcgatcctac catccactcg acacacccgc cagcggccgc
//
