LOCUS       sequence-306155-        4902 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 4902)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4902)
  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-m4134547
FEATURES             Location/Qualifiers
     source          1..4902
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      223..359
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    511..582
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(587..633)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    651..699
                     /note="synthetic polyadenylation signal"
     enhancer        780..1083
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1085..1287
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1238..1258
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1356..1365
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2028..2048
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2049..2075
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2128..2249
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2139..2158)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2193..2212
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2364..2410
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2440..2576
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2621..2692
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2918..3506)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2998..3017)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(4252..4320,4321..4902,1..210)
                     /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(4470..4489)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
       61 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
      121 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
      181 ataggtgcct cactgattaa gcattggtaa ccgattctag gtgcattggc gcagaaaaaa
      241 atgcctgatg cgacgctgcg cgtcttatac tcccacatat gccagattca gcaacggata
      301 cggcttcccc aacttgccca cttccatacg tgtcctcctt accagaaatt tatccttaag
      361 atcccgaatc gtttaaactc gactctggct ctatcgaatc tccgtcgttt cgagcttacg
      421 cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct
      481 tacctttttg gcagcgatcg cggctcccga catcttggac cattagctcc acaggtatct
      541 tcttccctct agtggtcata acagcagctt cagctacctc tcaattcaaa aaacccctca
      601 agacccgttt agaggcccca aggggttatg ctatcaatcg ttgcgttaca cacacaaaaa
      661 accaacacac atccatcttc gatggatagc gattttatta tctaactgct gatcgagtgt
      721 agccagatct agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
      781 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      841 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      901 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      961 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     1021 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     1081 atgctgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     1141 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     1201 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     1261 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatcag atctttgtcg
     1321 atcctaccat ccactcgaca cacccgccag cggccgccac catggcggac agcaacggca
     1381 ctattacagt cgaagagctt aagaaattac tcgagcagtg gaatttggta ataggattcc
     1441 tgtttctaac gtggatctgt ctgctgcaat tcgcatatgc caaccggaat agatttctct
     1501 acatcattaa gttgatcttc ctgtggctgc tttggcctgt gaccttggct tgctttgttc
     1561 tggccgccgt gtaccgcata aactggatta ccggtgggat cgctatcgca atggcgtgcc
     1621 tcgtgggcct tatgtggctg tcctatttta tagcctcatt ccgactcttt gctcgtacta
     1681 ggagtatgtg gtctttcaat cccgaaacaa atattttact aaacgtcccg ctgcatggga
     1741 ccattttaac aagaccattg ctagagagcg agctcgtgat cggcgcagtt atactgcggg
     1801 gacacctcag gatcgccgga caccatctgg gtcgctgtga tattaaagac ctgcccaagg
     1861 aaataacagt ggctacctcg cgtactcttt catactataa actgggagcc agccagagag
     1921 tggccgggga ttctggcttc gcagcatata gtcgatacag gatcgggaac tacaagctga
     1981 atacggacca ctcctcctct agtgataata ttgcattgct cgtacaggaa aacctttact
     2041 ttcagtcgta cccctatgac gtaccagatt atgcgtaagc tagcttgact gactgagata
     2101 cagcgtacct tcagctcaca gacatgataa gatacattga tgagtttgga caaaccacaa
     2161 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     2221 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     2281 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     2341 ttggcccatc tctatcggta tcgtagcata accccttggg gcctctaaac gggtcttgag
     2401 gggttttttg tgcccctcgg gccggattgc tatctaccgg cattggcgca gaaaaaaatg
     2461 cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca acggatacgg
     2521 cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat ccttaaggtc
     2581 gtcagctatc ctgcaggcga tctctcgatt tcgatcaaga cattccttta atggtctttt
     2641 ctggacacca ctaggggtca gaagtagttc atcaaacttt cttccctccc taatctcatt
     2701 ggttaccttg ggctatcgaa acttaattaa ccagtcaagt cagctacttg gcgagatcga
     2761 cttgtctggg tttcgactac gctcagaatt gcgtcagtca agttcgatct ggtccttgct
     2821 attgcacccg ttctccgatt acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa
     2881 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     2941 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3001 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3061 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3121 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3181 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3241 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3301 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     3361 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3421 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3481 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3541 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3601 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3661 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3721 tatttcgttc atccatagtt gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa
     3781 atcttcaaac ctttcgtccg atccatcttg caggctacct ctcgaacgaa ctatcgcaag
     3841 tctcttggcc ggccttgcgc cttggctatt gcttggcagc gcctatcgcc aggtattact
     3901 ccaatcccga atatccgaga tcgggatcac ccgagagaag ttcaacctac atcctcaatc
     3961 ccgatctatc cgagatccga ggaatatcga aatcggggcg cgcctggtgt accgagaacg
     4021 atcctctcag tgcgagtctc gacgatccat atcgttgctt ggcagtcagc cagtcggaat
     4081 ccagcttggg acccaggaag tccaatcgtc agatattgta ctcaagcctg gtcacggcag
     4141 cgtaccgatc tgtttaaacc tagatattga tagtctgatc ggtcaacgta taatcgagtc
     4201 ctagcttttg caaacatcta tcaagagaca ggatcagcag gaggctttcg catgagtatt
     4261 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     4321 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc gcgagtgggt
     4381 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgc
     4441 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     4501 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtat
     4561 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     4621 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat tggaggaccg
     4681 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     4741 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     4801 atggcaacaa ccttgcgtaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4861 cagttgatag actggatgga ggcggataaa gttgcaggac ca
//
