LOCUS       sequence-223956-        6222 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 6222)
  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 6222)
  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..6222
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     385..404
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     638..655
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    670..812
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(738..757)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     898..920
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(914..1105)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2417..3499)
                     /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"
     primer_bind     3461..3480
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3500..3577)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3783..3802
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3886..3905
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3886..3904
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3905..3929
                     /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             3944..3966
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3985..4002
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4012..4029
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             4535..4552
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4605..4623)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4619..4666
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(4841..4846,4847..4863,4864..4869))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(4896..4914)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4982..5086
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5087..5155,5156..5947)
                     /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(5305..5324)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6118..6222,1..484)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
       61 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
      121 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
      181 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
      241 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
      301 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
      361 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
      421 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
      481 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
      541 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
      601 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
      661 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
      721 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
      781 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
      841 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
      901 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
      961 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
     1021 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     1081 gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg
     1141 gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac
     1201 atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc
     1261 agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac
     1321 agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact
     1381 tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg
     1441 tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct
     1501 gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca
     1561 tgcgcacccg tggccaggac ccaacgctgc ccgagatgcg ccgcgtgcgg ctgctggaga
     1621 tggcggacgc gatggatatg ttctgccaag ggttggtttg cgcattcaca gttctccgca
     1681 agaattgatt ggctccaatt cttggagtgg tgaatccgtt agcgaggtgc cgccggcttc
     1741 cattcaggtc gaggtggccc ggctccatgc accgcgacgc aacgcgggga ggcagacaag
     1801 gtatagggcg gcgcctacaa tccatgccaa cccgttccat gtgctcgccg aggcggcata
     1861 aatcgccgtg acgatcagcg gtccagtgat cgaagttagg ctggtaagag ccgcgagcga
     1921 tccttgaagc tgtccctgat ggtcgtcatc tacctgcctg gacagcatgg cctgcaacgc
     1981 gggcatcccg atgccgccgg aagcgagaag aatcataatg gggaaggcca tccagcctcg
     2041 cgtcgcgaac gccagcaaga cgtagcccag cgcgtcggcc gccatgccgg cgataatggc
     2101 ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg
     2161 caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc
     2221 ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa
     2281 gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg
     2341 gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac
     2401 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     2461 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt
     2521 ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga
     2581 gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg
     2641 ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat
     2701 ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat
     2761 cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt
     2821 gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc
     2881 gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc
     2941 ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg
     3001 taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa
     3061 ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg
     3121 gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac
     3181 aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg
     3241 cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg
     3301 caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt
     3361 aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg
     3421 cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt
     3481 ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg
     3541 ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta
     3601 tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc
     3661 gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
     3721 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
     3781 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
     3841 acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc gaaattaata cgactcacta
     3901 taggggaatt gtgagcggat aacaattccc ctctagaaat aattttgttt aactttaaga
     3961 aggagatata ccatgggctg gagccatcat catcatcatc acagcagcgg cctggtgccg
     4021 cgcggcagcc atatgctcaa aggattcaag gagtttctcg cgcggggtaa tatcgtcgac
     4081 ctggctgtcg cggtggtaat cggcacagcg ttcacggcgt tggtcaccaa gttcaccgac
     4141 agcatcatta cgccgctgat caaccggatc ggcgtcaacg cacagtccga cgtcggcatc
     4201 ttgcggatcg gtatcggcgg tggtcagacc attgacttga acgtcttgtt gtcggcagcg
     4261 atcaactttt tcctgatcgc gttcgcggtg tacttcctag tcgtgctgcc ctacaacaca
     4321 ctacgcaaga aggggtaagt cgagcagccg ggcgacaccc aagtcgtgct gctcaccgaa
     4381 atccgcgatc tgctcgcgca aacgaacggg gactcgccgg ggaggcacgg cggccgtggg
     4441 acaccatcgc caaccgacgg gcctcgcgcg agcacagaat cgcaacccgg gtaagaattc
     4501 gagctccgtc gacaagcttg cggccgcact cgagcaccac caccaccacc actgagatcc
     4561 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     4621 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
     4681 tatatccgga tatcccgcaa gaggcccggc agtaccggca taaccaagcc tatgcctaca
     4741 gcatccaggg tgacggtgcc gaggatgacg atgagcgcat tgttagattt catacacggt
     4801 gcctgactgc gttagcaatt taactgtgat aaactaccgc attaaagctt atcgatgata
     4861 agctgtcaaa catgagaatt cttgaagacg aaagggcctc gtgatacgcc tatttttata
     4921 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     4981 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     5041 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     5101 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     5161 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     5221 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     5281 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     5341 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     5401 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5461 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     5521 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     5581 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc
     5641 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     5701 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     5761 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     5821 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     5881 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     5941 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     6001 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     6061 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6121 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6181 ggtggtttgt ttgccggatc aagagctacc aactcttttt cc
//
