LOCUS       sequence-300258-        4920 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 4920)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4920)
  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-m4134096
FEATURES             Location/Qualifiers
     source          1..4920
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             671..691
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             692..736
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     polyA_signal    789..910
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(800..819)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     854..873
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      1025..1071
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1101..1237
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1282..1353
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1579..2167)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1659..1678)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2913..2981,2982..3773)
                     /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(3131..3150)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3786..3922
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4074..4145
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4150..4196)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4214..4262
                     /note="synthetic polyadenylation signal"
     enhancer        4343..4646
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4648..4850
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4801..4821
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(4919..4920,1..8)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caccatggca gattccaacg gtactattac cgttgaagag cttaaaaagc tccttgaaca
       61 atggaaccta gtaataggtt tcctattcct tacatggatt tgtcttctac aatttgccta
      121 tgccaacagg aataggtttt tgtatataat taagttaatt ttcctctggc tgttatggcc
      181 agtaacttta gcttgttttg tgcttgctgc tgtttacaga ataaattgga tcaccggtgg
      241 aattgctatc gcaatggctt gtcttgtagg cttgatgtgg ctcagctact tcattgcttc
      301 tttcagactg tttgcgcgta cgcgttccat gtggtcattc aatccagaaa ctaacattct
      361 tctcaacgtg ccactccatg gcactattct gaccagaccg cttctagaaa gtgaactcgt
      421 aatcggagct gtgatccttc gtggacatct tcgtattgct ggacaccatc taggacgctg
      481 tgacatcaag gacctgccta aagaaatcac tgttgctaca tcacgaacgc tttcttatta
      541 caaattggga gcttcgcagc gtgtagcagg tgactcaggt tttgctgcat acagtcgcta
      601 caggattggc aactataaat taaacacaga ccattccagt agcagtgaca atattgcttt
      661 gcttgtacag gaaaaccttt actttcagtc gaaagaaacc gcggcagcta agttcgagag
      721 gcagcacatg gacagttaag ctagcttgac tgactgagat acagcgtacc ttcagctcac
      781 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
      841 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
      901 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
      961 ggaggttttt taaagcaagt aaaacctcta caaatgtggt attggcccat ctctatcggt
     1021 atcgtagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gtgcccctcg
     1081 ggccggattg ctatctaccg gcattggcgc agaaaaaaat gcctgatgcg acgctgcgcg
     1141 tcttatactc ccacatatgc cagattcagc aacggatacg gcttccccaa cttgcccact
     1201 tccatacgtg tcctccttac cagaaattta tccttaaggt cgtcagctat cctgcaggcg
     1261 atctctcgat ttcgatcaag acattccttt aatggtcttt tctggacacc actaggggtc
     1321 agaagtagtt catcaaactt tcttccctcc ctaatctcat tggttacctt gggctatcga
     1381 aacttaatta accagtcaag tcagctactt ggcgagatcg acttgtctgg gtttcgacta
     1441 cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc tattgcaccc gttctccgat
     1501 tacgagtttc atttaaatca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1561 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1621 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1681 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1741 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1801 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1861 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1921 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1981 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     2041 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2101 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2161 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2221 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2281 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2341 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2401 tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa aatcttcaaa cctttcgtcc
     2461 gatccatctt gcaggctacc tctcgaacga actatcgcaa gtctcttggc cggccttgcg
     2521 ccttggctat tgcttggcag cgcctatcgc caggtattac tccaatcccg aatatccgag
     2581 atcgggatca cccgagagaa gttcaaccta catcctcaat cccgatctat ccgagatccg
     2641 aggaatatcg aaatcggggc gcgcctggtg taccgagaac gatcctctca gtgcgagtct
     2701 cgacgatcca tatcgttgct tggcagtcag ccagtcggaa tccagcttgg gacccaggaa
     2761 gtccaatcgt cagatattgt actcaagcct ggtcacggca gcgtaccgat ctgtttaaac
     2821 ctagatattg atagtctgat cggtcaacgt ataatcgagt cctagctttt gcaaacatct
     2881 atcaagagac aggatcagca ggaggctttc gcatgagtat tcaacatttc cgtgtcgccc
     2941 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3001 aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa ctggatctca
     3061 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ctttccaatg atgagcactt
     3121 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     3181 gtcgccgcat acactattct cagaatgact tggttgagta ttcaccagtc acagaaaagc
     3241 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     3301 acactgcggc caacttactt ctgacaacga ttggaggacc gaaggagcta accgcttttt
     3361 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     3421 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca accttgcgta
     3481 aactattaac tggcgaacta cttactctag cttcccggca acagttgata gactggatgg
     3541 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3601 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3661 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3721 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc
     3781 taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca
     3841 tatgccagat tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc
     3901 cttaccagaa atttatcctt aagatcccga atcgtttaaa ctcgactctg gctctatcga
     3961 atctccgtcg tttcgagctt acgcgaacag ccgtggcgct catttgctcg tcgggcatcg
     4021 aatctcgtca gctatcgtca gcttaccttt ttggcagcga tcgcggctcc cgacatcttg
     4081 gaccattagc tccacaggta tcttcttccc tctagtggtc ataacagcag cttcagctac
     4141 ctctcaattc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctatcaa
     4201 tcgttgcgtt acacacacaa aaaaccaaca cacatccatc ttcgatggat agcgatttta
     4261 ttatctaact gctgatcgag tgtagccaga tctagtaatc aattacgggg tcattagttc
     4321 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     4381 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     4441 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     4501 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     4561 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     4621 acgtattagt catcgctatt accatgctga tgcggttttg gcagtacatc aatgggcgtg
     4681 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     4741 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
     4801 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga
     4861 accgtcagat cagatctttg tcgatcctac catccactcg acacacccgc cagcggccgc
//
