LOCUS       sequence-317525-        6809 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 6809)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 6809)
  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-m4133554
FEATURES             Location/Qualifiers
     source          1..6809
                     /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..1296
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1349..1470
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1360..1379)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1414..1433
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1585..1631
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1661..1797
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1842..1913
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(2139..2727)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2219..2238)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3227..4436
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     4273..4292
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4440..5039
                     /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(4440..4459)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4936..4956
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5066..5114
                     /note="synthetic polyadenylation signal"
     CDS             join(5380..5448,5449..6240)
                     /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(5598..5617)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      6253..6389
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6541..6612
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(6617..6663)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    6681..6729
                     /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 tttcagtcgt acccctatga cgtaccagat tatgcgtaag ctagcttgac tgactgagat
     1321 acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg acaaaccaca
     1381 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     1441 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     1501 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     1561 attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa cgggtcttga
     1621 ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc agaaaaaaat
     1681 gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg
     1741 gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta tccttaaggt
     1801 cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt aatggtcttt
     1861 tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc ctaatctcat
     1921 tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt ggcgagatcg
     1981 acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc
     2041 tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa aggccagcaa
     2101 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2161 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2221 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2281 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2341 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2401 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2461 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     2521 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     2581 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     2641 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     2701 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     2761 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     2821 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     2881 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     2941 ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa
     3001 aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga actatcgcaa
     3061 gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc caggtattac
     3121 tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta catcctcaat
     3181 cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggcc tccgcgccgg
     3241 gttttggcgc ctcccgcggg cgcccccctc gtcacggcga gcgctgccac gtcagacgaa
     3301 gggcgcagga gcgtcctgat ccttccgccc ggacgctcag gacagcggcc cgctgctcat
     3361 aagactcggc cttagaaccc cagtatcagc agaaggacat tttaggacgg gacttgggtg
     3421 actctagggc actggttttc tttccagaga gcggaacagg cgaggaaaag tagtcccttc
     3481 tcggcgattc tgcggaggga tctccgtggg gcggtgaacg ccgatgatta tataaggacg
     3541 cgccgggtgt ggcacagcta gttccgtcgc agccgggatt tgggtcgcgg ttcttgtttg
     3601 tggatcgctg tgatcgtcac ttggtgagta gcgggctgct gggctggccg gggctttcgt
     3661 ggccgccggg ccgctcggtg ggacggaagc gtgtggagag accgccaagg gctgtagtct
     3721 gggtccgcga gcaaggttgc cctgaactgg gggttggggg gagcgcagca aaatggcggc
     3781 tgttcccgag tcttgaatgg aagacgcttg tgaggcgggc tgtgaggtcg ttgaaacaag
     3841 gtggggggca tggtgggcgg caagaaccca aggtcttgag cccttcgcta atgcgggaaa
     3901 gctcttattc gggtgagatg ggctgggcac catctgggga ccctgacgtg aagtttgtca
     3961 ctgactggag aactcggttt gtcgtctgtt gcgggggcgg cagttatggc ggtgccgttg
     4021 ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg tcacccgttc
     4081 tgttggctta taatgcaggg tggggccacc tgccggtagg tgtgcggtag gcttttctcc
     4141 gtcgcaggac gcagggttcg ggcctagggt aggctctcct gaatcgacag gcgccggacc
     4201 tctggtgagg ggagggataa gtgaggcgtc agtttctttg gtcggtttta tgtacctatc
     4261 ttcttaagta gctgaagctc cggttttgaa ctatgcgctc ggggttggcg agtgtgtttt
     4321 gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt aattttcagt
     4381 gttagacttg taaattgtcc gctaaattct ggccgttttt ggcttttttg ttagacaaca
     4441 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac
     4501 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc
     4561 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
     4621 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
     4681 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
     4741 cccggctggc cgcgcagcaa cagatggagg gcctcctggc gccgcaccgg cccaaggagc
     4801 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
     4861 gcgccgtcgt gctccccgga gtggaggccg ccgagcgcgc cggggtgccc gccttcctgg
     4921 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     4981 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctagg
     5041 ctagtatgta agcctagtct tagataataa aatcgctatc catcgaagat ggatgtgtgt
     5101 tggttttttg tgtgtgtaac gctaggcgcg cctggtgtac cgagaacgat cctctcagtg
     5161 cgagtctcga cgatccatat cgttgcttgg cagtcagcca gtcggaatcc agcttgggac
     5221 ccaggaagtc caatcgtcag atattgtact caagcctggt cacggcagcg taccgatctg
     5281 tttaaaccta gatattgata gtctgatcgg tcaacgtata atcgagtcct agcttttgca
     5341 aacatctatc aagagacagg atcagcagga ggctttcgca tgagtattca acatttccgt
     5401 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5461 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta catcgaactg
     5521 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgctt tccaatgatg
     5581 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     5641 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtattc accagtcaca
     5701 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5761 agtgataaca ctgcggccaa cttacttctg acaacgattg gaggaccgaa ggagctaacc
     5821 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5881 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacc
     5941 ttgcgtaaac tattaactgg cgaactactt actctagctt cccggcaaca gttgatagac
     6001 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     6061 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     6121 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     6181 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     6241 ccgattctag gtgcattggc gcagaaaaaa atgcctgatg cgacgctgcg cgtcttatac
     6301 tcccacatat gccagattca gcaacggata cggcttcccc aacttgccca cttccatacg
     6361 tgtcctcctt accagaaatt tatccttaag atcccgaatc gtttaaactc gactctggct
     6421 ctatcgaatc tccgtcgttt cgagcttacg cgaacagccg tggcgctcat ttgctcgtcg
     6481 ggcatcgaat ctcgtcagct atcgtcagct tacctttttg gcagcgatcg cggctcccga
     6541 catcttggac cattagctcc acaggtatct tcttccctct agtggtcata acagcagctt
     6601 cagctacctc tcaattcaaa aaacccctca agacccgttt agaggcccca aggggttatg
     6661 ctatcaatcg ttgcgttaca cacacaaaaa accaacacac atccatcttc gatggatagc
     6721 gattttatta tctaactgct gatcgagtgt agccagatct agtaatcaat tacggggtca
     6781 ttagttcata gcccatatat ggagttccg
//
