LOCUS       sequence-313292-        6860 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 6860)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6860)
  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-m4134515
FEATURES             Location/Qualifiers
     source          1..6860
                     /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..1347
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     polyA_signal    1400..1521
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1411..1430)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1465..1484
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1636..1682
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1712..1848
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1893..1964
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2190..2778)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2270..2289)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3278..4487
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4324..4343
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4491..5090
                     /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(4491..4510)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4987..5007
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5117..5165
                     /note="synthetic polyadenylation signal"
     CDS             join(5431..5499,5500..6291)
                     /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(5649..5668)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6304..6440
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6592..6663
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6668..6714)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6732..6780
                     /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 ccatggcgga cagcaacggc
      601 actattacag tcgaagagct taagaaatta ctcgagcagt ggaatttggt aataggattc
      661 ctgtttctaa cgtggatctg tctgctgcaa ttcgcatatg ccaaccggaa tagatttctc
      721 tacatcatta agttgatctt cctgtggctg ctttggcctg tgaccttggc ttgctttgtt
      781 ctggccgccg tgtaccgcat aaactggatt accggtggga tcgctatcgc aatggcgtgc
      841 ctcgtgggcc ttatgtggct gtcctatttt atagcctcat tccgactctt tgctcgtact
      901 aggagtatgt ggtctttcaa tcccgaaaca aatattttac taaacgtccc gctgcatggg
      961 accattttaa caagaccatt gctagagagc gagctcgtga tcggcgcagt tatactgcgg
     1021 ggacacctca ggatcgccgg acaccatctg ggtcgctgtg atattaaaga cctgcccaag
     1081 gaaataacag tggctacctc gcgtactctt tcatactata aactgggagc cagccagaga
     1141 gtggccgggg attctggctt cgcagcatat agtcgataca ggatcgggaa ctacaagctg
     1201 aatacggacc actcctcctc tagtgataat attgcattgc tcgtacagga aaacctttac
     1261 tttcagtcga agaggcgatg gaaaaagaat tttatcgcgg tgagtgcagc caacagattc
     1321 aaaaagatat cctcgtctgg tgccctctaa gctagcttga ctgactgaga tacagcgtac
     1381 cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     1441 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     1501 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     1561 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tattggccca
     1621 tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     1681 tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc
     1741 gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca
     1801 acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg tcgtcagcta
     1861 tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt ttctggacac
     1921 cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca ttggttacct
     1981 tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc gacttgtctg
     2041 ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg ctattgcacc
     2101 cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca aaaggccagg
     2161 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2221 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2281 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2341 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2401 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2461 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2521 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2581 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     2641 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2701 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2761 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     2821 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     2881 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     2941 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3001 tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga aaatcttcaa
     3061 acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca agtctcttgg
     3121 ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta ctccaatccc
     3181 gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa tcccgatcta
     3241 tccgagatcc gaggaatatc gaaatcgggg cgcgcctggc ctccgcgccg ggttttggcg
     3301 cctcccgcgg gcgcccccct cgtcacggcg agcgctgcca cgtcagacga agggcgcagg
     3361 agcgtcctga tccttccgcc cggacgctca ggacagcggc ccgctgctca taagactcgg
     3421 ccttagaacc ccagtatcag cagaaggaca ttttaggacg ggacttgggt gactctaggg
     3481 cactggtttt ctttccagag agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt
     3541 ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt atataaggac gcgccgggtg
     3601 tggcacagct agttccgtcg cagccgggat ttgggtcgcg gttcttgttt gtggatcgct
     3661 gtgatcgtca cttggtgagt agcgggctgc tgggctggcc ggggctttcg tggccgccgg
     3721 gccgctcggt gggacggaag cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg
     3781 agcaaggttg ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga
     3841 gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc
     3901 atggtgggcg gcaagaaccc aaggtcttga gcccttcgct aatgcgggaa agctcttatt
     3961 cgggtgagat gggctgggca ccatctgggg accctgacgt gaagtttgtc actgactgga
     4021 gaactcggtt tgtcgtctgt tgcgggggcg gcagttatgg cggtgccgtt gggcagtgca
     4081 cccgtacctt tgggagcgcg cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt
     4141 ataatgcagg gtggggccac ctgccggtag gtgtgcggta ggcttttctc cgtcgcagga
     4201 cgcagggttc gggcctaggg taggctctcc tgaatcgaca ggcgccggac ctctggtgag
     4261 gggagggata agtgaggcgt cagtttcttt ggtcggtttt atgtacctat cttcttaagt
     4321 agctgaagct ccggttttga actatgcgct cggggttggc gagtgtgttt tgtgaagttt
     4381 tttaggcacc ttttgaaatg taatcatttg ggtcaatatg taattttcag tgttagactt
     4441 gtaaattgtc cgctaaattc tggccgtttt tggctttttt gttagacaac atgaccgagt
     4501 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg
     4561 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg
     4621 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     4681 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     4741 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     4801 ccgcgcagca acagatggag ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     4861 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     4921 tgctccccgg agtggaggcc gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     4981 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     5041 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctag gctagtatgt
     5101 aagcctagtc ttagataata aaatcgctat ccatcgaaga tggatgtgtg ttggtttttt
     5161 gtgtgtgtaa cgctaggcgc gcctggtgta ccgagaacga tcctctcagt gcgagtctcg
     5221 acgatccata tcgttgcttg gcagtcagcc agtcggaatc cagcttggga cccaggaagt
     5281 ccaatcgtca gatattgtac tcaagcctgg tcacggcagc gtaccgatct gtttaaacct
     5341 agatattgat agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc aaacatctat
     5401 caagagacag gatcagcagg aggctttcgc atgagtattc aacatttccg tgtcgccctt
     5461 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5521 gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt acatcgaact ggatctcaac
     5581 agcggtaaga tccttgagag ttttcgcccc gaagaacgct ttccaatgat gagcactttt
     5641 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5701 cgccgcatac actattctca gaatgacttg gttgagtatt caccagtcac agaaaagcat
     5761 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5821 actgcggcca acttacttct gacaacgatt ggaggaccga aggagctaac cgcttttttg
     5881 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5941 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa
     6001 ctattaactg gcgaactact tactctagct tcccggcaac agttgataga ctggatggag
     6061 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6121 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6181 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6241 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta accgattcta
     6301 ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata
     6361 tgccagattc agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct
     6421 taccagaaat ttatccttaa gatcccgaat cgtttaaact cgactctggc tctatcgaat
     6481 ctccgtcgtt tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa
     6541 tctcgtcagc tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg acatcttgga
     6601 ccattagctc cacaggtatc ttcttccctc tagtggtcat aacagcagct tcagctacct
     6661 ctcaattcaa aaaacccctc aagacccgtt tagaggcccc aaggggttat gctatcaatc
     6721 gttgcgttac acacacaaaa aaccaacaca catccatctt cgatggatag cgattttatt
     6781 atctaactgc tgatcgagtg tagccagatc tagtaatcaa ttacggggtc attagttcat
     6841 agcccatata tggagttccg
//
