Basic Vector Information
- Vector Name:
- pKL13
- Antibiotic Resistance:
- Ampicillin
- Length:
- 14206 bp
- Type:
- Cloning vector
- Replication origin:
- ori2
- Source/Author:
- Lam KN.
pKL13 vector Map
pKL13 vector Sequence
LOCUS 40924_26839 14206 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pKL13, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14206)
AUTHORS Lam KN.
TITLE Developing a Bacteroides system for function-based screening of DNA
from the human gut microbiome
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 14206)
AUTHORS Lam KN.
TITLE Direct Submission
JOURNAL Submitted (22-FEB-2016) Biology, University of Waterloo, 200
University Avenue West, Waterloo, Canada
REFERENCE 3 (bases 1 to 14206)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 14206)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(22-FEB-2016) Biology, University of Waterloo, 200 University Avenue
West, Waterloo, Canada"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..14206
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 288..304
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 311..329
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 332..337
/label=EcoRI site
/note="EcoRI site"
CDS complement(931..1620)
/codon_start=1
/gene="repA"
/product="replication protein"
/label=repA
/protein_id="AOP12487.1"
/translation="MILNRKIIFPLILFLWKIIVYFVENNISQKHIKMENKKAVKLTDF
QKNEENPFMKQAIEGIENHVVKKYKSNSGGDKRAVVALADTETGEVFKTSFIRQIEVDE
EQFTKLYLSNFAAFFDLSQAAIRVFGYFMTCMKPKNDLIIFNRKKCLEYTKYKTDKAVY
KGLAELVKAEIIARGPADNLWFINPLIVFNGDRVTFAKTYVRKKTLAAQKKEEAEKRQL
SLGFDEQ"
gene complement(931..1620)
/gene="repA"
/label=repA
CDS 3183..3983
/codon_start=1
/gene="ermF"
/product="erythromycin resistance"
/label=ermF
/protein_id="AOP12484.1"
/translation="MTKKKLPVRFTGQHFTIDKVLIKDAIRQANISNQDTVLDIGAGKG
FLTVHLLKIANNVVAIENDTALVEHLRKLFSDARNVQVVGCDFRNFAVPKFPFKVVSNI
PYGITSDIFKILMFESLGNFLGGSIVLQLEPTQKLFSRKLYNPYTVFYHTFFDLKLVYE
VGPESFLPPPTVKSALLNIKRKHLFFDFKFKAKYLAFISCLLEKPDLSVKTALKSIFRK
SQVRSISEKFGLNLNAQIVCLSPSQWLNCFLEMLEVVPEKFHPS"
gene 3183..3983
/gene="ermF"
/label=ermF
misc_feature 4133..4138
/label=EcoRI site
/note="EcoRI site"
misc_feature 4167..4174
/label=SgsI site
/note="SgsI site"
misc_feature 4183..4190
/label=PacI site
/note="PacI site"
regulatory complement(4191..4279)
/label=ilvGEDA transcriptional terminator
/note="ilvGEDA transcriptional terminator"
/regulatory_class="terminator"
primer_bind 4280..4310
/label=KL-JC103 sequencing primer
/note="KL-JC103 sequencing primer"
misc_feature complement(4311..4321)
/label=3-frame translational stop 2
/note="3-frame translational stop 2"
misc_feature 4322..4327
/label=NheI site
/note="NheI site"
misc_feature 4328..4333
/label=Eco72I site
/note="Eco72I site"
misc_feature 4334..4340
/label=Eco81I site
/note="Eco81I site"
protein_bind 4349..4365
/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)."
promoter complement(4373..4401)
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
misc_feature 4408..4414
/label=Eco81I site
/note="Eco81I site"
misc_feature 4415..4420
/label=BstBI site
/note="BstBI site"
CDS 4607..5407
/label=ApmR
/note="aminoglycoside 3-N-acetyltransferase type IV"
misc_feature 5476..5481
/label=BstBI site
/note="BstBI site"
misc_feature 5482..5487
/label=Eco72I site
/note="Eco72I site"
misc_feature 5488..5493
/label=NsiI site
/note="NsiI site"
misc_feature 5494..5504
/label=3-frame translational stop 1
/note="3-frame translational stop 1"
primer_bind complement(5505..5535)
/label=KL-JC102 sequencing primer
/note="KL-JC102 sequencing primer"
primer_bind complement(5506..5522)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
regulatory 5536..5617
/label=rnpB T1 transcriptional terminator
/note="rnpB T1 transcriptional terminator"
/regulatory_class="terminator"
misc_feature 5618..5624
/label=CpoI site
/note="CpoI site"
misc_feature 5633..5640
/label=SfaAI site
/note="SfaAI site"
oriT 5822..5931
/label=oriT
/note="incP origin of transfer"
CDS 5964..6332
/label=traJ
/note="oriT-recognizing protein"
primer_bind complement(6510..6526)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 6534..6550
/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)."
promoter complement(6558..6588)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(6603..6624)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(6847..7503)
/label=CmR
/note="chloramphenicol acetyltransferase"
promoter complement(7504..7606)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
CDS 7722..8069
/codon_start=1
/gene="redF"
/product="RedF"
/label=redF
/protein_id="AOP12489.1"
/translation="MERRNRRTGRTEKARIWEVTDRTVRTWIGEAVAAAAADGVTFSVP
VTPHTFRHSYAMHMLYAGIPLKVLQSLMGHKSISSTEVYTKVFALDVAARHRVQFAMPE
SDAVAMLKQLS"
gene 7722..8069
/gene="redF"
/label=redF
rep_origin 8464..9078
/label=oriV
/note="origin of replication for the bacterial F plasmid"
rep_origin 9154..9373
/label=ori2
/note="secondary origin of replication for the bacterial F
plasmid; also known as oriS"
CDS 9464..10216
/label=repE
/note="replication initiation protein for the bacterial F
plasmid"
misc_feature 10222..10472
/label=incC
/note="incompatibility region of the bacterial F plasmid"
CDS 10798..11970
/label=sopA
/note="partitioning protein for the bacterial F plasmid"
CDS 11973..12941
/label=sopB
/note="partitioning protein for the bacterial F plasmid"
misc_feature 13017..13490
/label=sopC
/note="centromere-like partitioning region of the bacterial
F plasmid"
misc_feature complement(13750..14148)
/label=cos
/note="lambda cos site; allows packaging into phage lambda
particles"
protein_bind complement(14166..14199)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
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