LOCUS       sequence-297077-        4427 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for SARS-CoV-2 M (membrane).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4427)
  TITLE     SARS-CoV-2 protein E. coli expression constructs, tagged, for 
            protein expression and/or purification
REFERENCE   2  (bases 1 to 4427)
  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-m4136454
FEATURES             Location/Qualifiers
     source          1..4427
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     14..33
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     162..181
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        162..180
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    181..205
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             220..242
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             253..270
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             589..609
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     terminator      1423..1454
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1455..1557
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(1558..1626,1627..2418)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1776..1795)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             2419..2610
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      2634..2661
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2673..3260)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2753..2772)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3282..3311)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     CDS             complement(join(3397..4427,1..52))
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc atattcacca
       61 ccctgaattg actgtcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt
      121 cgatactagt agcggccgga tggatctcga tcccgcgaaa ttaatacgac tcactatagg
      181 ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga
      241 gactcttcta tgcaccatca tcaccatcat ggtggttctg gtatgtctga ttctgaagtt
      301 aaccaagaag ctaaacctga ggttaagcca gaagtcaaac cagaaactca tattaacttg
      361 aaggtttctg atggttcttc tgaaatcttc ttcaagatca agaagaccac tccattgcgt
      421 agattgatgg aagctttcgc taaaagacaa ggtaaagaaa tggattcttt gagattcttg
      481 tacgacggta tcagaattca agctgatcaa actcctgaag atttggatat ggaggataat
      541 gacatcatcg aagctcatag agaacaaatt ggtggtggtg gttctggtga gaacctgtac
      601 tttcaaagcg ccgattcaaa cggtacgatt actgtagaag agcttaaaaa gctgttagaa
      661 caatggaatt tggtgatcgg gtttctgttc ctgacctgga tctgtctcct gcaattcgct
      721 tacgcgaacc gcaaccggtt tctgtatata atcaaattaa ttttcctgtg gctgctgtgg
      781 cccgtcaccc tggcatgctt cgttctggct gctgtttaca gaatcaattg gattactggc
      841 ggtatcgcaa tcgcgatggc atgcctagta ggcctgatgt ggctgtcgta ttttattgct
      901 tccttccgtc tgtttgcccg tacccgtagc atgtggtctt tcaacccaga aactaacatc
      961 ctgcttaacg ttcctctgca cggtacaatc ctgacccgcc cgctgctgga gagcgaactg
     1021 gtgattggcg ctgtcatctt acgtggtcat ctgcgtattg ctggccacca tttgggtcgc
     1081 tgtgacatca aagacttgcc gaaagaaatc actgttgcaa cctctcgtac actgtcctac
     1141 tacaaactgg gggcgagtca gcgtgttgcc ggcgattctg gattcgcggc ttacagccgc
     1201 tatcgcatcg gtaattacaa gctcaacact gaccacagct cctctagtga taacattgca
     1261 ctgctggtgc agtaaagaga atataaaaag ccagattatt aatccggctt ttttattatt
     1321 taggttctgt taagtaactg aacccaatgt cgttagtgac gcttacctct taagaggtca
     1381 ctgacctaac agcttgactc cagcgtaact ggactgcaat caactcaaaa gcctccgacc
     1441 ggaggctttt gacttgatcg gcacgtaaga ggttccaact ttcaccataa tgaaataaga
     1501 tcactaccgg gcgtattttt tgagttatcg agattttcag gagctaagga agctaaaatg
     1561 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     1621 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     1681 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt acgccccgaa
     1741 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     1801 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     1861 gagtactcac cagtcacaga aaagcatctc acggatggca tgacagtaag agaattatgc
     1921 agtgctgcca taaccatgag tgataacact gcggccaact tacttctggc aacgatcgga
     1981 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     2041 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     2101 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     2161 cggcaacaat taatagactg gatggaggcg gataaagttg caggatcact tctgcgctcg
     2221 gccctcccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     2281 ggtatcattg cagcactggg gccagatggt aagccctccc gcatcgtagt tatctacacg
     2341 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     2401 ctgattaagc attggtaagt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt
     2461 atcagaagcc agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag
     2521 gcagacatct gtgaatcgct tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg
     2581 cgtttcggtg atgacggtga aaacctctga tgagggccca aatgtaatca cctggctcac
     2641 cttcgggtgg gcctttctgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2701 catcacaaaa atcgatgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2761 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2821 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2881 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2941 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     3001 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     3061 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     3121 tttggtatct gcgctctgct gaagccagtt acctcggaaa aagagttggt agctcttgat
     3181 ccggcaaaca aaccaccgct ggtagcggtg gtctttttgt ttgcaagcag cagattacgc
     3241 gcagaaaaaa aggatctcaa gaagatcctt tgattttcta ccgaagaaag gcccacccgt
     3301 gaaggtgagc cagtgacttt ctgctatgga ggtcaggtat gatttaaatg gtcagtgcgg
     3361 cgcgccaaca aaaaacccct agccccccgt tttatatcac tgcccgcttt ccagtcggga
     3421 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     3481 attgggcgcc agggtggttt ttcttttcac cagtgagact ggcaacagct gattgccctt
     3541 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
     3601 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctatctt cggtatcgtc
     3661 gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
     3721 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
     3781 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
     3841 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
     3901 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
     3961 atgctccacg cccagtcgcg taccgtcctc atgggagaaa ataatactgt tgatgggtgt
     4021 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
     4081 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
     4141 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
     4201 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
     4261 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
     4321 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
     4381 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtct
//
