LOCUS       sequence-307278-        5430 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 5430)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 5430)
  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-m4133584
FEATURES             Location/Qualifiers
     source          1..5430
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    25..73
                     /note="synthetic polyadenylation signal"
     enhancer        154..457
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        459..661
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     612..632
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      730..739
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1402..1422
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1456..1533
                     /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"
     CDS             1561..1581
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    2030..2151
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2041..2060)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2095..2114
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2266..2312
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2342..2478
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2523..2594
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2820..3408)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2900..2919)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4154..4222,4223..5014)
                     /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(4372..4391)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      5027..5163
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    5315..5386
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(join(5391..5430,1..7))
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat cttcgatgga
       61 tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat caattacggg
      121 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      181 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      241 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      301 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
      361 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
      421 gcagtacatc tacgtattag tcatcgctat taccatgctg atgcggtttt ggcagtacat
      481 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      541 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
      601 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      661 tggtttagtg aaccgtcaga tcagatcttt gtcgatccta ccatccactc gacacacccg
      721 ccagcggccg ccaccatggc agattccaac ggtactatta ccgttgaaga gcttaaaaag
      781 ctccttgaac aatggaacct agtaataggt ttcctattcc ttacatggat ttgtcttcta
      841 caatttgcct atgccaacag gaataggttt ttgtatataa ttaagttaat tttcctctgg
      901 ctgttatggc cagtaacttt agcttgtttt gtgcttgctg ctgtttacag aataaattgg
      961 atcaccggtg gaattgctat cgcaatggct tgtcttgtag gcttgatgtg gctcagctac
     1021 ttcattgctt ctttcagact gtttgcgcgt acgcgttcca tgtggtcatt caatccagaa
     1081 actaacattc ttctcaacgt gccactccat ggcactattc tgaccagacc gcttctagaa
     1141 agtgaactcg taatcggagc tgtgatcctt cgtggacatc ttcgtattgc tggacaccat
     1201 ctaggacgct gtgacatcaa ggacctgcct aaagaaatca ctgttgctac atcacgaacg
     1261 ctttcttatt acaaattggg agcttcgcag cgtgtagcag gtgactcagg ttttgctgca
     1321 tacagtcgct acaggattgg caactataaa ttaaacacag accattccag tagcagtgac
     1381 aatattgctt tgcttgtaca ggaaaacctt tactttcagt cgggacggcg aatcccgggg
     1441 ctgataaatc cctggaaaag gagatggaag aagaacttta ttgcagtgtc tgcggctaac
     1501 cggttcaaaa agatcagttc aagcggcgcc ctagattacg acattcctac gaccgcctcc
     1561 gagaatctct atttccaggg tgaatttggc ttggcacaac atgacgaggc cgtcgataac
     1621 aagtttaata aagaacagca gaacgctttc tatgagatcc ttcacttacc aaatctgaac
     1681 gaagagcaac gcaacgcttt cattcagagc ctgaaagacg atccctcgca gtctgcaaat
     1741 ctcctggccg aagccaagaa gctgaatgac gcacaggctc caaaagttga taacaagttt
     1801 aacaaggagc agcagaatgc tttctacgag atattgcacc ttcctaatct gaatgaggaa
     1861 caaagaaatg catttatcca gtccctaaaa gacgatccat cacagagtgc caacctgctt
     1921 gcggaagcca aaaagctcaa tgatgcccag gccccgaaag tagacgcgaa ccatcaataa
     1981 gctagcttga ctgactgaga tacagcgtac cttcagctca cagacatgat aagatacatt
     2041 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     2101 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     2161 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
     2221 taaaacctct acaaatgtgg tattggccca tctctatcgg tatcgtagca taaccccttg
     2281 gggcctctaa acgggtcttg aggggttttt tgtgcccctc gggccggatt gctatctacc
     2341 ggcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact cccacatatg
     2401 ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt gtcctcctta
     2461 ccagaaattt atccttaagg tcgtcagcta tcctgcaggc gatctctcga tttcgatcaa
     2521 gacattcctt taatggtctt ttctggacac cactaggggt cagaagtagt tcatcaaact
     2581 ttcttccctc cctaatctca ttggttacct tgggctatcg aaacttaatt aaccagtcaa
     2641 gtcagctact tggcgagatc gacttgtctg ggtttcgact acgctcagaa ttgcgtcagt
     2701 caagttcgat ctggtccttg ctattgcacc cgttctccga ttacgagttt catttaaatc
     2761 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     2821 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     2881 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     2941 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3001 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3061 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3121 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3181 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3241 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3301 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     3361 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     3421 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     3481 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3541 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3601 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcatttaa atttccgaac
     3661 tctccaaggc cctcgtcgga aaatcttcaa acctttcgtc cgatccatct tgcaggctac
     3721 ctctcgaacg aactatcgca agtctcttgg ccggccttgc gccttggcta ttgcttggca
     3781 gcgcctatcg ccaggtatta ctccaatccc gaatatccga gatcgggatc acccgagaga
     3841 agttcaacct acatcctcaa tcccgatcta tccgagatcc gaggaatatc gaaatcgggg
     3901 cgcgcctggt gtaccgagaa cgatcctctc agtgcgagtc tcgacgatcc atatcgttgc
     3961 ttggcagtca gccagtcgga atccagcttg ggacccagga agtccaatcg tcagatattg
     4021 tactcaagcc tggtcacggc agcgtaccga tctgtttaaa cctagatatt gatagtctga
     4081 tcggtcaacg tataatcgag tcctagcttt tgcaaacatc tatcaagaga caggatcagc
     4141 aggaggcttt cgcatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     4201 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     4261 tcagttgggt gcgcgagtgg gttacatcga actggatctc aacagcggta agatccttga
     4321 gagttttcgc cccgaagaac gctttccaat gatgagcact tttaaagttc tgctatgtgg
     4381 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     4441 tcagaatgac ttggttgagt attcaccagt cacagaaaag catcttacgg atggcatgac
     4501 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4561 tctgacaacg attggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4621 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4681 tgacaccacg atgcctgtag caatggcaac aaccttgcgt aaactattaa ctggcgaact
     4741 acttactcta gcttcccggc aacagttgat agactggatg gaggcggata aagttgcagg
     4801 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4861 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4921 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4981 tgagataggt gcctcactga ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa
     5041 aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg
     5101 gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct
     5161 taagatcccg aatcgtttaa actcgactct ggctctatcg aatctccgtc gtttcgagct
     5221 tacgcgaaca gccgtggcgc tcatttgctc gtcgggcatc gaatctcgtc agctatcgtc
     5281 agcttacctt tttggcagcg atcgcggctc ccgacatctt ggaccattag ctccacaggt
     5341 atcttcttcc ctctagtggt cataacagca gcttcagcta cctctcaatt caaaaaaccc
     5401 ctcaagaccc gtttagaggc cccaaggggt
//
