pKL13 载体 (V005219#)

载体基本信息
载体名称 pKL13 载体抗性 Ampicillin
载体长度 14206 bp 载体类型 Cloning vector
载体来源 Lam KN.

pKL13 载体载体图谱

pKL1314206 bp7001400210028003500420049005600630070007700840091009800105001120011900126001330014000M13 fwdT7 promoterEcoRI siterepAermFEcoRI siteSgsI sitePacI siteilvGEDA transcriptional terminatorKL-JC103 sequencing primer3-frame translational stop 2NheI siteEco72I siteEco81I sitelac operatortac promoterEco81I siteBstBI siteApmRBstBI siteEco72I siteNsiI site3-frame translational stop 1KL-JC102 sequencing primerrnpB T1 transcriptional terminatorCpoI siteSfaAI siteoriTtraJM13 revlac operatorlac promoterCAP binding siteCmRcat promoterRedForiVori2repEincCsopAsopBsopCcosloxP

pKL13 载体载体序列

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  JOURNAL   Unpublished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            "Unpublished"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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