pKILHIS-1 vector (V005235)

LOCUS       40924_26724        3555 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pKILHIS-1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3555)
  AUTHORS   Van Reeth T, Dreze PL, Szpirer J, Szpirer C, Gabant P.
  TITLE     Positive selection vectors to generate fused genes for the 
            expression of his-tagged proteins
  JOURNAL   BioTechniques 25 (5), 898-904 (1998)
  PUBMED    9821593
REFERENCE   2  (bases 1 to 3555)
  AUTHORS   Van Reeth T, Gabant P, Dreze P, Szpirer J, Szpirer C.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-OCT-1997) Departement de Biologie Moleculaire 
            Laboratoire de Biologie du Developpement, Universite Libre de 
            Bruxelles, 67, Rue des Chevaux, 1640 Rhode Saint-Genese, Belgium
REFERENCE   3  (bases 1 to 3555)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3555)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "BioTechniques"; date: "1998"; volume: "25"; issue: "5"; pages: 
            "898-904"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (07-OCT-1997) Departement de Biologie Moleculaire Laboratoire de 
            Biologie du Developpement, Universite Libre de Bruxelles, 67, Rue 
            des Chevaux, 1640 Rhode Saint-Genese, Belgium"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3555
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        239..257
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     promoter        290..308
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             338..360
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             380..397
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             401..433
                     /codon_start=1
                     /label=T7 tag (gene 10 leader)
                     /note="leader peptide from bacteriophage T7 gene 10"
                     /translation="MASMTGGQQMG"
     misc_feature    458..470
                     /gene="ccdB"
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             626..925
                     /codon_start=1
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     terminator      1010..1057
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1153..1532
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(1583..2374)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     rep_origin      2786..3374
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"