LOCUS       Exported               12538 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector HKBS1, complete sequence.
ACCESSION   AF251497
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12538)
  AUTHORS   Hoang TT, Kutchma AJ, Becher A, Schweizer HP.
  TITLE     Integration-proficient plasmids for Pseudomonas aeruginosa: 
            site-specific integration and use for engineering of reporter and 
            expression strains
  JOURNAL   Plasmid 43 (1), 59-72 (2000)
  PUBMED    10610820
REFERENCE   2  (bases 1 to 12538)
  AUTHORS   Becher A, Schweizer HP.
  TITLE     Integration-proficient Pseudomonas aeruginosa vectors for isolation 
            of single copy chromosomal lacZ and lux gene fusions
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 12538)
  AUTHORS   Becher A, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-APR-2000) Microbiology, Colorado State University, 
            Fort Collins, CO 80523, USA
REFERENCE   4  (bases 1 to 12538)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12538
                     /organism="Cloning vector HKBS1"
                     /mol_type="other DNA"
                     /label=mini-CTX-lacZ
                     /note="mini-CTX-lacZ"
                     /db_xref="taxon:136151"
     protein_bind    238..285
                     /label=Flp recombinase binding site
                     /bound_moiety="Flp recombinase"
     misc_feature    495..521
                     /note="phage CTX attachment site; core sequence"
     regulatory      827..864
                     /regulatory_class="promoter"
                     /note="T3 promoter"
     promoter        827..845
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             complement(1565..2677)
                     /codon_start=1
                     /gene="luxE"
                     /product="LuxE"
                     /label=luxE
                     /note="component of fatty acid reductase complex"
                     /protein_id="AAG09983.1"
                     /translation="MTSYVDKQEITASSEIDDLIFSSDPLVWSYDEQEKIRKKLVLDAF
                     RNHYKHCREYRHYCQAHKVDDNITEIDDIPVFPTSVFKFTRLLTSQENEIESWFTSSGT
                     NGLKSQVARDRLSIERLLGSVSYGMKYVGSWFDHQIELVNLGPDRFNAHNIWFKYVMSL
                     VELLYPTTFTVTEERIDFVKTLNSLERIKNQGKDLCLIGSPYFIYLLCHYMKDKKISFS
                     GDKSLYIITGGGWKSYEKESLKRDDFNHLLFDTFNLSDISQIRDIFNQVELNTCFFEDE
                     MQRKHVPPWVYARALDPETLKPVPDGTPGLMSYMDASATSYPAFIVTDDVGIISREYGK
                     YPGVLVEILRRVNTRTQKGCALSLTEAFDS"
     gene            complement(1565..2677)
                     /gene="luxE"
                     /label=luxE
     CDS             complement(2856..3839)
                     /codon_start=1
                     /gene="luxB"
                     /product="luciferase"
                     /label=luxB
                     /protein_id="AAG09984.1"
                     /translation="MKFGLFFLNFINSTTVQEQSIVRMQEITEYVDKLNFEQILVYENH
                     FSDNGVVGAPLTVSGFLLGLTEKIKIGSLNHIITTHHPVAIAEEACLLDQLSEGRFILG
                     FSDCEKKDEMHFFNRPVEYQQQLFEECYEIINDALTTGYCNPDNDFYSFPKISVNPHAY
                     TPGGPRKYVTATSHHIVEWAAKKGIPLIFKWDDSNDVRYEYAERYKAVADKYDVDLSEI
                     DHQLMILVNYNEDSNKAKQETRAFISDYVLEMHPNENFENKLEEIIAENAVGNYTECIT
                     AAKLAIEKCGAKSVLLSFEPMNDLMSQKNVINIVDDNIKKYHMEYT"
     gene            complement(2856..3839)
                     /gene="luxB"
                     /label=luxB
     CDS             complement(3854..4936)
                     /codon_start=1
                     /gene="luxA"
                     /product="luciferase"
                     /label=luxA
                     /protein_id="AAG09985.1"
                     /translation="MKFGNFLLTYQPPQFSQTEVMKRLVKLGRISEECGFDTVWLLEHH
                     FTEFGLLGNPYVAAAYLLGATKKLNVGTAAIVLPTAHPVRQLEDVNLLDQMSKGRFRFG
                     ICRGLYNKDFRVFGTDMNNSRALAECWYGLIKNGMTEGYMEADNEHIKFHKVKVNPAAY
                     SRGGAPVYVVAESASTTEWAAQFGLPMILSWIINTNEKKAQLELYNEVAQEYGHDIHNI
                     DHCLSYITSVDHDSIKAKEICRKFLGHWYDSYVNATTIFDDSDQTRGYDFNKGQWRDFV
                     LKGHKDTNRRIDYSYEINPVGTPQECIDIIQKDIDATGISNICCGFEANGTVDEIIASM
                     KLFQSDVMPFLKEKQRSLLY"
     gene            complement(3854..4936)
                     /gene="luxA"
                     /label=luxA
     CDS             complement(4985..5908)
                     /codon_start=1
                     /gene="luxD"
                     /product="acyltransferase"
                     /label=luxD
                     /protein_id="AAG09986.1"
                     /translation="MENESKYKTIDHVICVEGNKKIHVWETLPEENSPKRKNAIIIASG
                     FARRMDHFAGLAEYLSRNGFHVIRYDSLHHVGLSSGTIDEFTMSIGKQSLLAVVDWLTT
                     RKINNFGMLASSLSARIAYASLSEINASFLITAVGVVNLRYSLERALGFDYLSLPINEL
                     PDNLDFEGHKLGAEVFARDCLDFGWEDLASTINNMMYLDIPFIAFTANNDNWVKQDEVI
                     TLLSNIRSNRCKIYSLLGSSHDLSENLVVLRNFYQSVTKAAIAMDNDHLDIDVDITEPS
                     FEHLTIATVNERRMRIEIENQAISLS"
     gene            complement(4985..5908)
                     /gene="luxD"
                     /label=luxD
     CDS             complement(5920..7362)
                     /codon_start=1
                     /gene="lucC"
                     /product="fatty acid reductase"
                     /label=lucC
                     /protein_id="AAG09987.1"
                     /translation="MTKKISFIINGQVEIFPESDDLVQSINFGDNSVYLPILNDSHVKN
                     IIDCNGNNELRLHNIVNFLYTVGQRWKNEEYSRRRTYIRDLKKYMGYSEEMAKLEANWI
                     SMILCSKGGLYDVVENELGSRHIMDEWLPQDESYVRAFPKGKSVHLLAGNVPLSGIMSI
                     LRAILTKNQCIIKTSSTDPFTANALALSFIDVDPNHPITRSLSVIYWPHQGDTSLAKEI
                     MRHADVIVAWGGPDAINWAVEHAPSYADVIKFGSKKSLCIIDNPVDLTSAATGAAHDVC
                     FYDQRACFSAQNIYYMGNHYEEFKLALIEKLNLYAHILPNAKKDFDEKAAYSLVQKESL
                     FAGLKVEVDIHQRWMIIESNAGVEFNQPLGRCVYLHHVDNIEQILPYVQKNKTQTISIF
                     PWESSFKYRDALALKGAERIVEAGMNNIFRVGGSHDGMRPLQRLVTYISHERPSNYTAK
                     DVAVEIEQTRFLEEDKFLVFVP"
     gene            complement(5920..7362)
                     /gene="lucC"
                     /label=lucC
     misc_feature    7822..7893
                     /label=multiple cloning site
                     /note="multiple cloning site"
     primer_bind     complement(7860..7876)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      7901..7920
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     promoter        complement(7902..7920)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    8094..8141
                     /label=Flp recombinase binding site
                     /bound_moiety="Flp recombinase"
     oriT            8341..8600
     oriT            8348..8456
                     /note="incP origin of transfer"
     CDS             complement(8615..9784)
                     /codon_start=1
                     /gene="int"
                     /product="integrase"
                     /label=int
                     /note="modified integrase"
                     /protein_id="AAG09988.1"
                     /translation="MADGVEVRGKRIRIYFRYQGELCRESIPGDATPENIANAERLAGI
                     INYEIKQGVFSYSRHFPDSPRVKSNTLGHYIDLWLDIKRNQIAASGFRGYTSRVETHIR
                     PRWGDSQADSIDHLDIQDWVQNTLMPKLHNKTVREIVSNLRQIFRLYRTRNRSAHDPTD
                     GIVITLPDADDPDPFTREEIDLILGTETARIGELNLAEFMIWSGPRVSEAIALAWEDVD
                     LDTGTVVFRRARVRSQYKVTKTRRSTRKVQLLAPALRALQQQAKLTRRLPPVQIEVIDR
                     DNRTRKPQRVRFVFHNSASGAAYSTSDTLRNGWWHGHLRNAGVRSRGPNQCRHTFASQM
                     LSSGIATPEWIADQMGHTSTAMIFKHYAKWISKDGPDIVGLLNQALKLS"
     gene            complement(8615..9784)
                     /gene="int"
                     /label=int
     rep_origin      complement(9881..10469)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(11291..12481)
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline resistance protein"
                     /label=tet
                     /note="modified tet"
                     /protein_id="AAG09989.1"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     gene            complement(11291..12481)
                     /gene="tet"
                     /label=tet
     promoter        complement(12529..19)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"