LOCUS       sequence-228502-        6218 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  M. tuberculosis MscL (delta49) in pET19b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6218)
  AUTHORS   Herrera N, Maksaev G, Haswell ES, Rees DC
  TITLE     Elucidating a role for the cytoplasmic domain in the Mycobacterium 
            tuberculosis mechanosensitive channel of large conductance.
  JOURNAL   Sci Rep. 2018 Oct 1;8(1):14566. doi: 10.1038/s41598-018-32536-6.
  PUBMED    30275500
REFERENCE   2  (bases 1 to 6218)
  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"; journalName: "Sci Rep."; 
            date: "2018-10-1"; volume: "8(1):14566. doi"; pages: " 
            10.1038/s41598-018-32536-6"
COMMENT     Sequence Label: MtMscL delta49
FEATURES             Location/Qualifiers
     source          1..6218
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             19..36
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             46..63
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             569..586
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(639..657)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      653..700
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(875..880,881..897,898..903))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(930..948)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1016..1120
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1121..1189,1190..1981)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1339..1358)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2152..2740
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2641..2660
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2894..2911
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2926..3065
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2991..3010)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3151..3173
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3167..3358)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5713..5732
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5752..5829)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6035..6054
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6138..6157
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6138..6156
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    6157..6181
                     /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             6196..6218
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 tataccatgg gctggagcca tcatcatcat catcacagca gcggcctggt gccgcgcggc
       61 agccatatgc tcaaaggatt caaggagttt ctcgcgcggg gtaatatcgt cgacctggct
      121 gtcgcggtgg taatcggcac agcgttcacg gcgttggtca ccaagttcac cgacagcatc
      181 attacgccgc tgatcaaccg gatcggcgtc aacgcacagt ccgacgtcgg catcttgcgg
      241 atcggtatcg gcggtggtca gaccattgac ttgaacgtct tgttgtcggc agcgatcaac
      301 tttttcctga tcgcgttcgc ggtgtacttc ctagtcgtgc tgccctacaa cacactacgc
      361 aagaaggggt aagtcgagca gccgggcgac acccaagtcg tgctgctcac cgaaatccgc
      421 gatctgctcg cgcaaacgaa cggggactcg ccggggaggc acggcggccg tgggacacca
      481 tcgccaaccg acgggcctcg cgcgagcaca gaatcgcaac ccgggtaaga attcgagctc
      541 cgtcgacaag cttgcggccg cactcgagca ccaccaccac caccactgag atccggctgc
      601 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
      661 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc
      721 cggatatccc gcaagaggcc cggcagtacc ggcataacca agcctatgcc tacagcatcc
      781 agggtgacgg tgccgaggat gacgatgagc gcattgttag atttcataca cggtgcctga
      841 ctgcgttagc aatttaactg tgataaacta ccgcattaaa gcttatcgat gataagctgt
      901 caaacatgag aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa
      961 tgtcatgata ataatggttt cttagaggtc aggtggcact tttcggggaa atgtgcgcgg
     1021 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     1081 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     1141 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     1201 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     1261 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     1321 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
     1381 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     1441 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     1501 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     1561 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     1621 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac
     1681 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     1741 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     1801 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     1861 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     1921 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     1981 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     2041 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     2101 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     2161 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     2221 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     2281 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     2341 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     2401 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     2461 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     2521 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     2581 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     2641 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     2701 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     2761 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     2821 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     2881 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     2941 tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc
     3001 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     3061 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3121 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     3181 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     3241 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     3301 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg
     3361 atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga
     3421 ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg
     3481 gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc
     3541 agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc
     3601 agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac
     3661 ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc
     3721 ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc
     3781 ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggccag gacccaacgc
     3841 tgcccgagat gcgccgcgtg cggctgctgg agatggcgga cgcgatggat atgttctgcc
     3901 aagggttggt ttgcgcattc acagttctcc gcaagaattg attggctcca attcttggag
     3961 tggtgaatcc gttagcgagg tgccgccggc ttccattcag gtcgaggtgg cccggctcca
     4021 tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg gcggcgccta caatccatgc
     4081 caacccgttc catgtgctcg ccgaggcggc ataaatcgcc gtgacgatca gcggtccaat
     4141 gatcgaagtt aggctggtaa gagccgcgag cgatccttga agctgtccct gatggtcgtc
     4201 atctacctgc ctggacagca tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag
     4261 aagaatcata atggggaagg ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc
     4321 cagcgcgtcg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc
     4381 gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag
     4441 gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc
     4501 cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt
     4561 catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg
     4621 agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc
     4681 tttccagtcg ggaaactgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4741 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
     4801 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
     4861 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
     4921 ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt
     4981 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
     5041 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
     5101 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
     5161 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
     5221 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
     5281 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
     5341 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
     5401 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
     5461 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
     5521 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
     5581 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     5641 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     5701 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     5761 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     5821 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     5881 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     5941 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     6001 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     6061 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
     6121 tctcgatccc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattc
     6181 ccctctagaa ataattttgt ttaactttaa gaaggaga
//
