LOCUS       sequence-317731-        6800 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 6800)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6800)
  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-m4133550
FEATURES             Location/Qualifiers
     source          1..6800
                     /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"
     promoter        3218..4427
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4264..4283
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4431..5030
                     /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(4431..4450)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4927..4947
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5057..5105
                     /note="synthetic polyadenylation signal"
     CDS             join(5371..5439,5440..6231)
                     /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(5589..5608)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6244..6380
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6532..6603
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6608..6654)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6672..6720
                     /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 ccatggcaga ttccaacggt
      601 actattaccg ttgaagagct taaaaagctc cttgaacaat ggaacctagt aataggtttc
      661 ctattcctta catggatttg tcttctacaa tttgcctatg ccaacaggaa taggtttttg
      721 tatataatta agttaatttt cctctggctg ttatggccag taactttagc ttgttttgtg
      781 cttgctgctg tttacagaat aaattggatc accggtggaa ttgctatcgc aatggcttgt
      841 cttgtaggct tgatgtggct cagctacttc attgcttctt tcagactgtt tgcgcgtacg
      901 cgttccatgt ggtcattcaa tccagaaact aacattcttc tcaacgtgcc actccatggc
      961 actattctga ccagaccgct tctagaaagt gaactcgtaa tcggagctgt gatccttcgt
     1021 ggacatcttc gtattgctgg acaccatcta ggacgctgtg acatcaagga cctgcctaaa
     1081 gaaatcactg ttgctacatc acgaacgctt tcttattaca aattgggagc ttcgcagcgt
     1141 gtagcaggtg actcaggttt tgctgcatac agtcgctaca ggattggcaa ctataaatta
     1201 aacacagacc attccagtag cagtgacaat attgctttgc ttgtacagga 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 cgcgcctggc ctccgcgccg ggttttggcg
     3241 cctcccgcgg gcgcccccct cgtcacggcg agcgctgcca cgtcagacga agggcgcagg
     3301 agcgtcctga tccttccgcc cggacgctca ggacagcggc ccgctgctca taagactcgg
     3361 ccttagaacc ccagtatcag cagaaggaca ttttaggacg ggacttgggt gactctaggg
     3421 cactggtttt ctttccagag agcggaacag gcgaggaaaa gtagtccctt ctcggcgatt
     3481 ctgcggaggg atctccgtgg ggcggtgaac gccgatgatt atataaggac gcgccgggtg
     3541 tggcacagct agttccgtcg cagccgggat ttgggtcgcg gttcttgttt gtggatcgct
     3601 gtgatcgtca cttggtgagt agcgggctgc tgggctggcc ggggctttcg tggccgccgg
     3661 gccgctcggt gggacggaag cgtgtggaga gaccgccaag ggctgtagtc tgggtccgcg
     3721 agcaaggttg ccctgaactg ggggttgggg ggagcgcagc aaaatggcgg ctgttcccga
     3781 gtcttgaatg gaagacgctt gtgaggcggg ctgtgaggtc gttgaaacaa ggtggggggc
     3841 atggtgggcg gcaagaaccc aaggtcttga gcccttcgct aatgcgggaa agctcttatt
     3901 cgggtgagat gggctgggca ccatctgggg accctgacgt gaagtttgtc actgactgga
     3961 gaactcggtt tgtcgtctgt tgcgggggcg gcagttatgg cggtgccgtt gggcagtgca
     4021 cccgtacctt tgggagcgcg cgccctcgtc gtgtcgtgac gtcacccgtt ctgttggctt
     4081 ataatgcagg gtggggccac ctgccggtag gtgtgcggta ggcttttctc cgtcgcagga
     4141 cgcagggttc gggcctaggg taggctctcc tgaatcgaca ggcgccggac ctctggtgag
     4201 gggagggata agtgaggcgt cagtttcttt ggtcggtttt atgtacctat cttcttaagt
     4261 agctgaagct ccggttttga actatgcgct cggggttggc gagtgtgttt tgtgaagttt
     4321 tttaggcacc ttttgaaatg taatcatttg ggtcaatatg taattttcag tgttagactt
     4381 gtaaattgtc cgctaaattc tggccgtttt tggctttttt gttagacaac atgaccgagt
     4441 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg
     4501 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg
     4561 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg
     4621 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag
     4681 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg
     4741 ccgcgcagca acagatggag ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt
     4801 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg
     4861 tgctccccgg agtggaggcc gccgagcgcg ccggggtgcc cgccttcctg gagacctccg
     4921 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg
     4981 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctag gctagtatgt
     5041 aagcctagtc ttagataata aaatcgctat ccatcgaaga tggatgtgtg ttggtttttt
     5101 gtgtgtgtaa cgctaggcgc gcctggtgta ccgagaacga tcctctcagt gcgagtctcg
     5161 acgatccata tcgttgcttg gcagtcagcc agtcggaatc cagcttggga cccaggaagt
     5221 ccaatcgtca gatattgtac tcaagcctgg tcacggcagc gtaccgatct gtttaaacct
     5281 agatattgat agtctgatcg gtcaacgtat aatcgagtcc tagcttttgc aaacatctat
     5341 caagagacag gatcagcagg aggctttcgc atgagtattc aacatttccg tgtcgccctt
     5401 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     5461 gtaaaagatg ctgaagatca gttgggtgcg cgagtgggtt acatcgaact ggatctcaac
     5521 agcggtaaga tccttgagag ttttcgcccc gaagaacgct ttccaatgat gagcactttt
     5581 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5641 cgccgcatac actattctca gaatgacttg gttgagtatt caccagtcac agaaaagcat
     5701 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5761 actgcggcca acttacttct gacaacgatt ggaggaccga aggagctaac cgcttttttg
     5821 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5881 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac cttgcgtaaa
     5941 ctattaactg gcgaactact tactctagct tcccggcaac agttgataga ctggatggag
     6001 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6061 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6121 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     6181 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta accgattcta
     6241 ggtgcattgg cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata
     6301 tgccagattc agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct
     6361 taccagaaat ttatccttaa gatcccgaat cgtttaaact cgactctggc tctatcgaat
     6421 ctccgtcgtt tcgagcttac gcgaacagcc gtggcgctca tttgctcgtc gggcatcgaa
     6481 tctcgtcagc tatcgtcagc ttaccttttt ggcagcgatc gcggctcccg acatcttgga
     6541 ccattagctc cacaggtatc ttcttccctc tagtggtcat aacagcagct tcagctacct
     6601 ctcaattcaa aaaacccctc aagacccgtt tagaggcccc aaggggttat gctatcaatc
     6661 gttgcgttac acacacaaaa aaccaacaca catccatctt cgatggatag cgattttatt
     6721 atctaactgc tgatcgagtg tagccagatc tagtaatcaa ttacggggtc attagttcat
     6781 agcccatata tggagttccg
//
