LOCUS       sequence-295925-        4893 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 4893)
  TITLE     SARS-CoV-2 protein Mammalian expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4893)
  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-m4133720
FEATURES             Location/Qualifiers
     source          1..4893
                     /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..709
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    762..883
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(773..792)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     827..846
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      998..1044
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1074..1210
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    1255..1326
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(1552..2140)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1632..1651)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(2886..2954,2955..3746)
                     /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(3104..3123)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3759..3895
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    4047..4118
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(4123..4169)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    4187..4235
                     /note="synthetic polyadenylation signal"
     enhancer        4316..4619
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4621..4823
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4774..4794
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(4892..4893,1..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 gcaccatcac catcaccatt aagctagctt
      721 gactgactga gatacagcgt accttcagct cacagacatg ataagataca ttgatgagtt
      781 tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc
      841 tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat
      901 tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct
      961 ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct tggggcctct
     1021 aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta ccggcattgg
     1081 cgcagaaaaa aatgcctgat gcgacgctgc gcgtcttata ctcccacata tgccagattc
     1141 agcaacggat acggcttccc caacttgccc acttccatac gtgtcctcct taccagaaat
     1201 ttatccttaa ggtcgtcagc tatcctgcag gcgatctctc gatttcgatc aagacattcc
     1261 tttaatggtc ttttctggac accactaggg gtcagaagta gttcatcaaa ctttcttccc
     1321 tccctaatct cattggttac cttgggctat cgaaacttaa ttaaccagtc aagtcagcta
     1381 cttggcgaga tcgacttgtc tgggtttcga ctacgctcag aattgcgtca gtcaagttcg
     1441 atctggtcct tgctattgca cccgttctcc gattacgagt ttcatttaaa tcatgtgagc
     1501 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1561 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1621 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1681 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1741 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1801 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1861 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1921 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     1981 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2041 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2101 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2161 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2221 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2281 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2341 ctcagcgatc tgtctatttc gttcatccat agttgcattt aaatttccga actctccaag
     2401 gccctcgtcg gaaaatcttc aaacctttcg tccgatccat cttgcaggct acctctcgaa
     2461 cgaactatcg caagtctctt ggccggcctt gcgccttggc tattgcttgg cagcgcctat
     2521 cgccaggtat tactccaatc ccgaatatcc gagatcggga tcacccgaga gaagttcaac
     2581 ctacatcctc aatcccgatc tatccgagat ccgaggaata tcgaaatcgg ggcgcgcctg
     2641 gtgtaccgag aacgatcctc tcagtgcgag tctcgacgat ccatatcgtt gcttggcagt
     2701 cagccagtcg gaatccagct tgggacccag gaagtccaat cgtcagatat tgtactcaag
     2761 cctggtcacg gcagcgtacc gatctgttta aacctagata ttgatagtct gatcggtcaa
     2821 cgtataatcg agtcctagct tttgcaaaca tctatcaaga gacaggatca gcaggaggct
     2881 ttcgcatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     2941 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3001 gtgcgcgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3061 gccccgaaga acgctttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     3121 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3181 acttggttga gtattcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3241 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3301 cgattggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3361 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3421 cgatgcctgt agcaatggca acaaccttgc gtaaactatt aactggcgaa ctacttactc
     3481 tagcttcccg gcaacagttg atagactgga tggaggcgga taaagttgca ggaccacttc
     3541 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3601 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3661 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3721 gtgcctcact gattaagcat tggtaaccga ttctaggtgc attggcgcag aaaaaaatgc
     3781 ctgatgcgac gctgcgcgtc ttatactccc acatatgcca gattcagcaa cggatacggc
     3841 ttccccaact tgcccacttc catacgtgtc ctccttacca gaaatttatc cttaagatcc
     3901 cgaatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag cttacgcgaa
     3961 cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg tcagcttacc
     4021 tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag gtatcttctt
     4081 ccctctagtg gtcataacag cagcttcagc tacctctcaa ttcaaaaaac ccctcaagac
     4141 ccgtttagag gccccaaggg gttatgctat caatcgttgc gttacacaca caaaaaacca
     4201 acacacatcc atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc
     4261 agatctagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     4321 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     4381 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     4441 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     4501 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     4561 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgc
     4621 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     4681 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     4741 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     4801 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatcagatct ttgtcgatcc
     4861 taccatccac tcgacacacc cgccagcggc cgc
//
