LOCUS       Exported               11285 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pJAK14-Blue, complete sequence.
ACCESSION   JX310866
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11285)
  AUTHORS   Kornacki JA.
  TITLE     A family of broad-host-range vectors for expression of cloned genes 
            in Gram-negative bacteria
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 11285)
  AUTHORS   Xu K, Figurski DH.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-JUL-2012) Microbiology and Immunology, Columbia 
            University, 701W, 168th Street, Room 1514A, NYC, NY 10027, USA
REFERENCE   3  (bases 1 to 11285)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11285
                     /organism="Cloning vector pJAK14-Blue"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /label=kanamycin resistant
                     /note="kanamycin resistant"
                     /db_xref="taxon:1233322"
     regulatory      1..158
                     /regulatory_class="terminator"
                     /note="rrnB_terminator"
     terminator      121..207
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      124..167
                     /regulatory_class="terminator"
                     /note="rrnB_T1_terminator"
     regulatory      299..326
                     /regulatory_class="terminator"
                     /note="rrnB_T2_terminator"
     terminator      299..326
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        345..436
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      367..395
                     /regulatory_class="promoter"
                     /note="ampicillin promoter"
     mobile_element  648..2852
                     /mobile_element_type="transposon:transposon:mini-Tn5 Km"
     CDS             1446..2240
                     /codon_start=1
                     /product="neomycin-kanamycin phosphotransferase type II"
                     /label=neomycin-kanamycin phosphotransferase type II
                     /protein_id="AFS32160.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     rep_origin      3861..4255
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     oriT            4597..4684
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     CDS             5923..6894
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein B of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepB
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDAPTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGHDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     CDS             7405..8244
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein A of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepA
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     CDS             8231..9082
                     /codon_start=1
                     /gene="repC"
                     /product="replication protein C of the broad-host-range 
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /label=RSF1010 RepC
                     /translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TAECHVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRKRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     regulatory      9244..9274
                     /regulatory_class="promoter"
                     /note="P5 promoter"
     CDS             complement(9502..10317)
                     /codon_start=1
                     /product="LacI"
                     /label=LacI
                     /note="represses the expression of downstream lacZ-alpha 
                     gene and relives the repression when IPTG is added"
                     /protein_id="AFS32161.1"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSR
                     ADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPA
                     LFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKY
                     LTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLR
                     VGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTTWTACCNSLRARR"
     promoter        complement(10441..10518)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     promoter        10748..10776
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     regulatory      10750..10778
                     /regulatory_class="promoter"
                     /note="tac promoter"
     protein_bind    10784..10800
                     /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)."
     misc_feature    10790..10812
                     /label=M13_pUC_rev_primer
                     /note="M13_pUC_rev_primer"
     CDS             10831..11196
                     /codon_start=1
                     /product="LacZ-alpha"
                     /label=LacZ-alpha
                     /note="contains MCS region and provides Blue/White 
                     screening function"
                     /protein_id="AFS32162.1"
                     /translation="MTMITPSAQLTLTKGNKSWVPGPPSRSTVSISLISNSCSPGDPLV
                     LERPPPRWSSNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWKL"
     promoter        10856..10874
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    10887..10994
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     10904..10920
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10954..10970)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(11003..11021)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(11031..11047)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    11237..11254
                     /note="pBAD_rev_primer; pTrcHis_rev_primer"