pMTL-SC7215 vector (V004427)

Basic Vector Information

Plasmid vector pMTL-SC7215, used for allele exchange in C. difficile R20291. Note that pMTL-SC7215 and pMTL-SC7315 are identical except that they have the pBP1 and pCB102 Gram-positive replicons, respectively (between AscI and FseI restriction sites).

Vector Name:: pMTL-SC7215

Antibiotic Resistance:: Chloramphenicol

Length:: 6778 bp

Type:: C. difficile

Replication origin:: ori

Source/Author:: Cartman ST, Kelly ML, Heeg D, Heap JT, Minton NP.

Growth Strain(s):: DH5alpha

Growth Temperature:: 37℃

pMTL-SC7215 vector Map

Copy Sequence View Map

pMTL-SC7215 vector Sequence

LOCUS       V004427                 6778 bp    DNA     circular SYN 04-JAN-2024
DEFINITION  Exported.
ACCESSION   V004427
VERSION     V004427
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6778)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6778
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     63..79
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(254..278)
                     /label="lac operator"
                     /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             297..305
                     /label="Shine-Dalgarno sequence"
                     /note="full consensus sequence for ribosome-binding sites
                     upstream of start codons in E. coli; complementary to a
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             318..1598
                     /label="codA"
                     /note="E. coli cytosine deaminase"
     primer_bind     complement(1723..1739)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             2449..3504
                     /codon_start=1
                     /gene="repA"
                     /product="plasmid replication protein"
                     /label="repA"
                     /protein_id="ACR43886.1"
                     /translation="MTNYNQKNENKQEVKTALEKCTDKKRKNPRFTSYIAPLTTKKNIE
                     RTSTCGDYLFMLSDADLEHFKLHKGNFCGNRFCPMCSWRLACKDSLEISILMEHLRKEE
                     NKEFIFLTLTTPNVKSYDLNYSIKQYNKSFKKLMERKEVKDITKGYIRKLEVTYQKEKY
                     ITKDLWKIKKDYYQKKGLEIGDLEPNFDTYNPHFHVVIAVNKSYFTDKNYYINRERWLE
                     LWKFATKDDSITQVDVRKAKINDYKEVYELAKYSAKDTDYLISRPVFEIFYKALKGKQV
                     LVFSGFFKDAHKLYKQGKLDVYKKKDEIKYVYIVYYNWCKKQYEKTRIRELTEDEKEEL
                     NQDLIDEIEID"
     gene            2449..3504
                     /gene="repA"
                     /label="repA"
     CDS             complement(3595..4098)
                     /codon_start=1
                     /product="putative plasmid replication protein"
                     /label="putative plasmid replication protein"
                     /note="orf2"
                     /protein_id="ACR43887.1"
                     /translation="MKFKKIILYIFIVIIPLSITGCSINQITTPVPKNYLDNIVKKYVG
                     PEGVQPSFGGKTFYDYKIIDSEKSKNIIKIYLYYIGQEYYIEDDKLVEGTGSASFATVT
                     IKKENNNYKLVSYEVPKSPAQEDTVKIFPKSVRKKAIHANTPSLKNIEKEAETYFKKEL
                     KTKN"
     CDS             4422..5042
                     /gene="catP"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from Clostridium
                     perfringens. Accession#: P26826"
     rep_origin      5222..5810
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     oriT            6131..6240
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             6273..6641
                     /label="traJ"
                     /note="oriT-recognizing protein"

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