LOCUS       sequence-313277-        4917 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 4917)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4917)
  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-m4134535
FEATURES             Location/Qualifiers
     source          1..4917
                     /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..1311
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    1364..1485
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1375..1394)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1429..1448
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1600..1646
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1676..1812
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1857..1928
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2154..2742)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2234..2253)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3488..3556,3557..4348)
                     /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(3706..3725)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4361..4497
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4649..4720
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4725..4771)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4789..4837
                     /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 tttcagtcgg ggaaacctat acccaatcca cttctcggct tagattctac ctaagctagc
     1321 ttgactgact gagatacagc gtaccttcag ctcacagaca tgataagata cattgatgag
     1381 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     1441 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     1501 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     1561 ctctacaaat gtggtattgg cccatctcta tcggtatcgt agcataaccc cttggggcct
     1621 ctaaacgggt cttgaggggt tttttgtgcc cctcgggccg gattgctatc taccggcatt
     1681 ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat
     1741 tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa
     1801 atttatcctt aaggtcgtca gctatcctgc aggcgatctc tcgatttcga tcaagacatt
     1861 cctttaatgg tcttttctgg acaccactag gggtcagaag tagttcatca aactttcttc
     1921 cctccctaat ctcattggtt accttgggct atcgaaactt aattaaccag tcaagtcagc
     1981 tacttggcga gatcgacttg tctgggtttc gactacgctc agaattgcgt cagtcaagtt
     2041 cgatctggtc cttgctattg cacccgttct ccgattacga gtttcattta aatcatgtga
     2101 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2161 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2221 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2281 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2341 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2401 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2461 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2521 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2581 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2641 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2701 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2761 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     2821 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     2881 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     2941 atctcagcga tctgtctatt tcgttcatcc atagttgcat ttaaatttcc gaactctcca
     3001 aggccctcgt cggaaaatct tcaaaccttt cgtccgatcc atcttgcagg ctacctctcg
     3061 aacgaactat cgcaagtctc ttggccggcc ttgcgccttg gctattgctt ggcagcgcct
     3121 atcgccaggt attactccaa tcccgaatat ccgagatcgg gatcacccga gagaagttca
     3181 acctacatcc tcaatcccga tctatccgag atccgaggaa tatcgaaatc ggggcgcgcc
     3241 tggtgtaccg agaacgatcc tctcagtgcg agtctcgacg atccatatcg ttgcttggca
     3301 gtcagccagt cggaatccag cttgggaccc aggaagtcca atcgtcagat attgtactca
     3361 agcctggtca cggcagcgta ccgatctgtt taaacctaga tattgatagt ctgatcggtc
     3421 aacgtataat cgagtcctag cttttgcaaa catctatcaa gagacaggat cagcaggagg
     3481 ctttcgcatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     3541 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     3601 gggtgcgcga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     3661 tcgccccgaa gaacgctttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     3721 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     3781 tgacttggtt gagtattcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     3841 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     3901 aacgattgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     3961 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     4021 cacgatgcct gtagcaatgg caacaacctt gcgtaaacta ttaactggcg aactacttac
     4081 tctagcttcc cggcaacagt tgatagactg gatggaggcg gataaagttg caggaccact
     4141 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     4201 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     4261 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     4321 aggtgcctca ctgattaagc attggtaacc gattctaggt gcattggcgc agaaaaaaat
     4381 gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg
     4441 gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta tccttaagat
     4501 cccgaatcgt ttaaactcga ctctggctct atcgaatctc cgtcgtttcg agcttacgcg
     4561 aacagccgtg gcgctcattt gctcgtcggg catcgaatct cgtcagctat cgtcagctta
     4621 cctttttggc agcgatcgcg gctcccgaca tcttggacca ttagctccac aggtatcttc
     4681 ttccctctag tggtcataac agcagcttca gctacctctc aattcaaaaa acccctcaag
     4741 acccgtttag aggccccaag gggttatgct atcaatcgtt gcgttacaca cacaaaaaac
     4801 caacacacat ccatcttcga tggatagcga ttttattatc taactgctga tcgagtgtag
     4861 ccagatctag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccg
//
