LOCUS       sequence-357012-        6824 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 6824)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6824)
  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-m4134317
FEATURES             Location/Qualifiers
     source          1..6824
                     /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"
     promoter        3242..4451
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4288..4307
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4455..5054
                     /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(4455..4474)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4951..4971
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5081..5129
                     /note="synthetic polyadenylation signal"
     CDS             join(5395..5463,5464..6255)
                     /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(5613..5632)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6268..6404
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6556..6627
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6632..6678)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6696..6744
                     /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 tggcctccgc gccgggtttt ggcgcctccc gcgggcgccc ccctcgtcac ggcgagcgct
     3301 gccacgtcag acgaagggcg caggagcgtc ctgatccttc cgcccggacg ctcaggacag
     3361 cggcccgctg ctcataagac tcggccttag aaccccagta tcagcagaag gacattttag
     3421 gacgggactt gggtgactct agggcactgg ttttctttcc agagagcgga acaggcgagg
     3481 aaaagtagtc ccttctcggc gattctgcgg agggatctcc gtggggcggt gaacgccgat
     3541 gattatataa ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg ggatttgggt
     3601 cgcggttctt gtttgtggat cgctgtgatc gtcacttggt gagtagcggg ctgctgggct
     3661 ggccggggct ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc
     3721 caagggctgt agtctgggtc cgcgagcaag gttgccctga actgggggtt ggggggagcg
     3781 cagcaaaatg gcggctgttc ccgagtcttg aatggaagac gcttgtgagg cgggctgtga
     3841 ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga acccaaggtc ttgagccctt
     3901 cgctaatgcg ggaaagctct tattcgggtg agatgggctg ggcaccatct ggggaccctg
     3961 acgtgaagtt tgtcactgac tggagaactc ggtttgtcgt ctgttgcggg ggcggcagtt
     4021 atggcggtgc cgttgggcag tgcacccgta cctttgggag cgcgcgccct cgtcgtgtcg
     4081 tgacgtcacc cgttctgttg gcttataatg cagggtgggg ccacctgccg gtaggtgtgc
     4141 ggtaggcttt tctccgtcgc aggacgcagg gttcgggcct agggtaggct ctcctgaatc
     4201 gacaggcgcc ggacctctgg tgaggggagg gataagtgag gcgtcagttt ctttggtcgg
     4261 ttttatgtac ctatcttctt aagtagctga agctccggtt ttgaactatg cgctcggggt
     4321 tggcgagtgt gttttgtgaa gttttttagg caccttttga aatgtaatca tttgggtcaa
     4381 tatgtaattt tcagtgttag acttgtaaat tgtccgctaa attctggccg tttttggctt
     4441 ttttgttaga caacatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
     4501 tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
     4561 ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
     4621 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
     4681 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
     4741 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggagggcctc ctggcgccgc
     4801 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg
     4861 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggccgccgag cgcgccgggg
     4921 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
     4981 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
     5041 agcccggtgc ctaggctagt atgtaagcct agtcttagat aataaaatcg ctatccatcg
     5101 aagatggatg tgtgttggtt ttttgtgtgt gtaacgctag gcgcgcctgg tgtaccgaga
     5161 acgatcctct cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg
     5221 aatccagctt gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg
     5281 cagcgtaccg atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga
     5341 gtcctagctt ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgagt
     5401 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5461 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg
     5521 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5581 cgctttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     5641 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5701 tattcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5761 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gattggagga
     5821 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5881 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5941 gcaatggcaa caaccttgcg taaactatta actggcgaac tacttactct agcttcccgg
     6001 caacagttga tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     6061 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     6121 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     6181 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     6241 attaagcatt ggtaaccgat tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg
     6301 ctgcgcgtct tatactccca catatgccag attcagcaac ggatacggct tccccaactt
     6361 gcccacttcc atacgtgtcc tccttaccag aaatttatcc ttaagatccc gaatcgttta
     6421 aactcgactc tggctctatc gaatctccgt cgtttcgagc ttacgcgaac agccgtggcg
     6481 ctcatttgct cgtcgggcat cgaatctcgt cagctatcgt cagcttacct ttttggcagc
     6541 gatcgcggct cccgacatct tggaccatta gctccacagg tatcttcttc cctctagtgg
     6601 tcataacagc agcttcagct acctctcaat tcaaaaaacc cctcaagacc cgtttagagg
     6661 ccccaagggg ttatgctatc aatcgttgcg ttacacacac aaaaaaccaa cacacatcca
     6721 tcttcgatgg atagcgattt tattatctaa ctgctgatcg agtgtagcca gatctagtaa
     6781 tcaattacgg ggtcattagt tcatagccca tatatggagt tccg
//
