LOCUS       Exported               10295 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCD341, complete sequence.
ACCESSION   AF053122
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10295)
  AUTHORS   Dehio M, Knorre A, Lanz C, Dehio C.
  TITLE     Construction of versatile high-level expression vectors for 
            Bartonella henselae and the use of green fluorescent protein as a 
            new expression marker
  JOURNAL   Gene 215 (2), 223-229 (1998)
  PUBMED    9714815
REFERENCE   2  (bases 1 to 10295)
  AUTHORS   Dehio C, Dehio M, Knorre A, Lanz C.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-MAR-1998) Infektionsbiologie, Max-Planck-Institut fuer
            Biologie, Spemannstrasse 34, Tuebingen D-72076, Germany
REFERENCE   3  (bases 1 to 10295)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     Name: SYNPCD341.
FEATURES             Location/Qualifiers
     source          1..10295
                     /organism="Cloning vector pCD341"
                     /mol_type="other DNA"
                     /db_xref="taxon:74542"
     primer_bind     complement(37..53)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      392..478
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      570..597
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        616..707
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1158..1260
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1750..2544
                     /codon_start=1
                     /gene="neo"
                     /product="neomycin phosphotransferase"
                     /label=neo
                     /protein_id="AAC35766.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            1750..2544
                     /gene="neo"
                     /label=neo
     rep_origin      3236..3630
                     /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)"
     CDS             complement(3657..3941)
                     /codon_start=1
                     /gene="mobC"
                     /product="conjugational DNA transfer protein"
                     /label=mobC
                     /protein_id="AAC35767.1"
                     /translation="MVKGSNKAADRLAKLEEQRARINAEIQRERAREQQQERKNETRRK
                     VLVGAMILAKVNSSEWPEDRLMAAMDAYLERDHDRALFGLPPRQKDEPG"
     gene            complement(3657..3941)
                     /gene="mobC"
                     /label=mobC
     misc_feature    3894..4051
                     /function="origin of conjugational DNA transfer"
     oriT            3972..4059
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     CDS             4140..6269
                     /codon_start=1
                     /gene="repB/mobA"
                     /product="RepB/MobA"
                     /label=repB/mobA
                     /protein_id="AAC35768.1"
                     /translation="MAIYHLTAKTGSRSGGQSARAKADYIQREGKYARDMDEVLHAESG
                     HMPEFVERPADYWDAADLYERANGRLFKEVEFALPVELTLDQQKALASEFAQHLTGAER
                     LPYTLAIHAGGGENPHCHLMISERINDGIERPAAQWFKRYNGKTPEKGGAQKTEALKPK
                     AWLEQTREAWADHANRALERAGHDARIDHRTLEAQGIERLPGVHLGPNVVEMEGRGIRT
                     DRADVALNIDTANAQIIDLQEYREAIDHERNRQSEEIQRHQRVSGADRTAGPEHGDTGR
                     RSPAGHEPDQAGQRGAGGGVAESPAPDRGGMGGAGQRVAGGSRRGEQRRAERPERVAGV
                     ALEAMANRDAGFHDAYGGAADRIVALARPDATDNRGRLDLAALGGPMKNDRTLQAIGRQ
                     LKAMGCERFDIGVRDAPTGQMMNREWSAAEVLQNTPWLKRMNAQGNDVYIRPAEQERHG
                     LVLVDDLSEFDLDDMKAEGREPALVVETSPKNYQAWVKVADAAGGELRGQIARTLASEY
                     DADPASADSRHYGRLAGFTNRKDKHTTRAGYQPWVLLRESKGKTATAGPALVQQAGQQI
                     EQAQRQQEKARRLASLELPERQLSRHRRTALDEYRSEMAGLVKRFGHDLSKCDFIAAQK
                     LASRGRSAEEIGKAMAEASPALAERKPGHEADYIERTVSKVMGLPSVQLARAELARAPA
                     PRQRGMDRGGPDFSM"
     gene            4140..6269
                     /gene="repB/mobA"
                     /label=repB/mobA
     CDS             4888..5301
                     /codon_start=1
                     /gene="mobB"
                     /product="conjugal DNA transfer protein"
                     /label=mobB
                     /protein_id="AAC35769.1"
                     /translation="MNAIDRVKKSRGINELAEQIEPLAQSMATLADEARQVMSQTKQAS
                     EAQAAEWLKAQRQTGAAWVELAKELREVAAEVSSAAQSARSASRGWHWKLWLTVMLASM
                     MPTVVLLIASLLLLDLTPLTTEDGSIWLRLVAR"
     gene            4888..5301
                     /gene="mobB"
                     /label=mobB
     CDS             5298..6269
                     /codon_start=1
                     /gene="repB"
                     /product="primase"
                     /label=repB
                     /protein_id="AAC35770.1"
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDAPTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGHDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     gene            5298..6269
                     /gene="repB"
                     /label=repB
     CDS             6330..6542
                     /codon_start=1
                     /product="unknown"
                     /label=unknown
                     /note="E"
                     /protein_id="AAC35771.1"
                     /translation="MEYEKSASGSVYLIKSDKGYWLPGGFGYTSNKAEAGRFSVADMAS
                     LNLDGCTLSLFREDKPFGPGKFLGD"
     CDS             6544..6750
                     /codon_start=1
                     /gene="cac"
                     /product="Cac"
                     /function="regulates repC and repA gene expression"
                     /label=cac
                     /protein_id="AAC35772.1"
                     /translation="MKDQKDKQTGDLLASPDAVRQARYAERMKAKGMRQRKFWLTDDEY
                     EALRECLEELRAAQGGGSDPASA"
     gene            6544..6750
                     /gene="cac"
                     /label=cac
     CDS             6780..7619
                     /codon_start=1
                     /gene="repA"
                     /product="DNA helicase"
                     /label=repA
                     /protein_id="AAC35773.1"
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     gene            6780..7619
                     /gene="repA"
                     /label=repA
     CDS             7606..8457
                     /codon_start=1
                     /gene="repC"
                     /product="replication initiator protein"
                     /label=repC
                     /protein_id="AAC35774.1"
                     /translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TAECHVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRKRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     gene            7606..8457
                     /gene="repC"
                     /label=repC
     CDS             complement(8877..9692)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /protein_id="AAC35776.1"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSR
                     ADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPA
                     LFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKY
                     LTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLR
                     VGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTTWTACCNSLRARR"
     gene            complement(8877..9692)
                     /gene="lacI"
                     /label=lacI
     promoter        complement(9816..9893)
                     /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        10123..10151
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     regulatory      10124..10152
                     /regulatory_class="promoter"
                     /note="tac"
     protein_bind    10159..10175
                     /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        10205..10235
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     regulatory      10207..10235
                     /regulatory_class="promoter"
                     /note="lacUV5"
     protein_bind    10243..10259
                     /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)."
     primer_bind     10267..10283
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             10279..181
                     /codon_start=1
                     /gene="lacZ"
                     /product="alpha-peptide"
                     /label=lacZ
                     /protein_id="AAC35775.1"
                     /translation="MTMITNSRGSVDLQPSLALAVVLQRRDWENPGVTQLNRLAAHPPF
                     ASWLIAKRPAPIALPNSCAA"
     gene            10279..181
                     /gene="lacZ"
                     /label=lacZ