pMTL575555 vector (V016037)

Basic Vector Information

Vector Name:
pMTL575555
Length:
7196 bp
Type:
Cloning vector
Replication origin:
ori
Source/Author:
Lau MSH., Sheng L, Zhang Y, Minton NP.

pMTL575555 vector Map

pMTL5755557196 bp30060090012001500180021002400270030003300360039004200450048005100540057006000630066006900St1Cas9lactate dehydrogenase promoter and RBSglyceraldehyde 3-phosphate dehydrogenase promoterrrnB T1 terminatorrrnB T2 terminatorM13 fwdrepBkanamycin nucleotidyltransferaseori

pMTL575555 vector Sequence

LOCUS       62056_17560        7196 bp DNA     circular SYN 28-JUL-2021
DEFINITION  Cloning vector pMTL575555, complete sequence.
ACCESSION   MZ182078
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7196)
  AUTHORS   Lau MSH., Sheng L, Zhang Y, Minton NP.
  TITLE     Development of a Suite of Tools for Genome Editing in 
            Parageobacillus thermoglucosidasius and Their Use to Identify the 
            Potential of a Native Plasmid in the Generation of Stable Engineered
            Strains
  JOURNAL   ACS Synth Biol 10 (7), 1739-1749 (2021)
  PUBMED    34197093
REFERENCE   2  (bases 1 to 7196)
  AUTHORS   Lau MSH., Sheng L, Zhang Y, Minton NP.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-MAY-2021) Biodiscovery Institute, School of Life 
            Sciences, University of Nottingham, Science Road, University Park, 
            Nottingham, Nottinghamshire NG7 2RD, United Kingdom
REFERENCE   3  (bases 1 to 7196)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol"; date: "2021"; volume: "10"; issue: "7"; pages: "1739-1749"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (10-MAY-2021) Biodiscovery Institute, School of Life Sciences, 
            University of Nottingham, Science Road, University Park, Nottingham,
            Nottinghamshire NG7 2RD, United Kingdom"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7196
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(12..3374)
                     /label=St1Cas9
                     /note="Cas9 endonuclease from the Streptococcus
                     thermophilus Type II CRISPR/Cas system (Esvelt et al., 
                     2013)"
     regulatory      complement(3375..3559)
                     /label=lactate dehydrogenase promoter and RBS
                     /note="lactate dehydrogenase promoter and RBS"
                     /regulatory_class="promoter"
     regulatory      3566..3656
                     /label=glyceraldehyde 3-phosphate dehydrogenase promoter
                     /note="glyceraldehyde 3-phosphate dehydrogenase promoter"
                     /regulatory_class="promoter"
     terminator      3794..3880
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3972..3999
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     primer_bind     complement(4009..4025)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4461..5462
                     /label=repB
                     /note="RepB replication protein"
     CDS             5619..6389
                     /codon_start=1
                     /transl_table=11
                     /product="kanamycin nucleotidyltransferase"
                     /label=kanamycin nucleotidyltransferase
                     /protein_id="QXV87316.1"
                     /translation="MRIVNGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGR
                     QTDGPYSDIEMMCVMSTEEAEFSHEWTTGEWKVEVNFYSEEILLDYASQVESDWPLTHG
                     QFFSILPIYDSGGYLEKVYQTAKSVEAQKFHDAICALIVEELFEYAGKWRNIRVQGPTT
                     FLPSLTVQVAMAGAMLIGLHHRICYTTSASVLTEAVKQSDLPSGYDHLCQFVMSGQLSD
                     SEKLLESLENFWNGIQEWTERHGYIVDVSKRIPF"
     rep_origin      6397..6980
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"

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