LOCUS       Exported                3555 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pKILHIS-1, complete sequence.
ACCESSION   AF028837
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3555
                     /organism="Cloning vector pKILHIS-1"
                     /host="Escherichia coli"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:74553"
     regulatory      95..203
                     /regulatory_class="promoter"
                     /note="Lac"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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"
     regulatory      290..309
                     /regulatory_class="promoter"
                     /note="T7"
     promoter        290..308
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             338..360
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     gene            368..1059
                     /gene="ccdB"
                     /label=ccdB
     CDS             368..928
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB protein"
                     /label=ccdB
                     /note="CcdB fusion protein"
                     /protein_id="AAC78278.1"
                     /translation="MRGSHHHHHHGMASMTGGQQMGRDLYDDDDIGSVDLAVVLQRRDW
                     ENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLYVRQFKVYTYKRESRYRLFVD
                     VQSDIIDTPGRRMVIPLASARLLSDKVSRELYPVVHIGDESWRMMTTDMASVPVSVIGE
                     EVADLSHRENDIKNAINLMFWGI"
     CDS             380..397
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             401..433
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     misc_feature    458..470
                     /gene="ccdB"
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             626..928
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     regulatory      962..1059
                     /regulatory_class="terminator"
                     /gene="ccdB"
     terminator      1010..1057
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1153..1532
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      1154..1565
                     /label=f1
                     /note="f1"
     CDS             complement(1580..2374)
                     /codon_start=1
                     /product="kanamycin marker protein"
                     /label=kanamycin marker protein
                     /protein_id="AAC78279.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     rep_origin      2761..3432
                     /label=ColE1
                     /note="ColE1"
     rep_origin      2786..3374
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"