LOCUS       sequence-313285-        4893 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 4893)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4893)
  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-m4134513
FEATURES             Location/Qualifiers
     source          1..4893
                     /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..1287
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1340..1461
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1351..1370)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1405..1424
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1576..1622
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1652..1788
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1833..1904
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2130..2718)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2210..2229)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(3464..3532,3533..4324)
                     /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(3682..3701)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4337..4473
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4625..4696
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4701..4747)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4765..4813
                     /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 tttcagtcgc accatcacca tcaccattaa gctagcttga ctgactgaga tacagcgtac
     1321 cttcagctca cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     1381 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     1441 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     1501 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tattggccca
     1561 tctctatcgg tatcgtagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     1621 tgtgcccctc gggccggatt gctatctacc ggcattggcg cagaaaaaaa tgcctgatgc
     1681 gacgctgcgc gtcttatact cccacatatg ccagattcag caacggatac ggcttcccca
     1741 acttgcccac ttccatacgt gtcctcctta ccagaaattt atccttaagg tcgtcagcta
     1801 tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt ttctggacac
     1861 cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca ttggttacct
     1921 tgggctatcg aaacttaatt aaccagtcaa gtcagctact tggcgagatc gacttgtctg
     1981 ggtttcgact acgctcagaa ttgcgtcagt caagttcgat ctggtccttg ctattgcacc
     2041 cgttctccga ttacgagttt catttaaatc atgtgagcaa aaggccagca aaaggccagg
     2101 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     2161 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     2221 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     2281 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2341 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2401 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2461 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2521 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     2581 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2641 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2701 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     2761 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     2821 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     2881 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     2941 tcatccatag ttgcatttaa atttccgaac tctccaaggc cctcgtcgga aaatcttcaa
     3001 acctttcgtc cgatccatct tgcaggctac ctctcgaacg aactatcgca agtctcttgg
     3061 ccggccttgc gccttggcta ttgcttggca gcgcctatcg ccaggtatta ctccaatccc
     3121 gaatatccga gatcgggatc acccgagaga agttcaacct acatcctcaa tcccgatcta
     3181 tccgagatcc gaggaatatc gaaatcgggg cgcgcctggt gtaccgagaa cgatcctctc
     3241 agtgcgagtc tcgacgatcc atatcgttgc ttggcagtca gccagtcgga atccagcttg
     3301 ggacccagga agtccaatcg tcagatattg tactcaagcc tggtcacggc agcgtaccga
     3361 tctgtttaaa cctagatatt gatagtctga tcggtcaacg tataatcgag tcctagcttt
     3421 tgcaaacatc tatcaagaga caggatcagc aggaggcttt cgcatgagta ttcaacattt
     3481 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3541 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcgcgagtgg gttacatcga
     3601 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gctttccaat
     3661 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     3721 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt attcaccagt
     3781 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     3841 catgagtgat aacactgcgg ccaacttact tctgacaacg attggaggac cgaaggagct
     3901 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     3961 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4021 aaccttgcgt aaactattaa ctggcgaact acttactcta gcttcccggc aacagttgat
     4081 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4141 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4201 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4261 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4321 gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt
     4381 atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca
     4441 tacgtgtcct ccttaccaga aatttatcct taagatcccg aatcgtttaa actcgactct
     4501 ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc
     4561 gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg atcgcggctc
     4621 ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt cataacagca
     4681 gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt
     4741 tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat cttcgatgga
     4801 tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat caattacggg
     4861 gtcattagtt catagcccat atatggagtt ccg
//
