LOCUS       Exported                6755 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Integration vector mini-CTX2, complete sequence.
ACCESSION   AF140577
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6755)
  AUTHORS   Hoang TT, Kutchma AJ, Becher A, Schweizer HP.
  TITLE     A site-specific gene integration system for Pseudomonas aeruginosa
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6755)
  AUTHORS   Hoang TT, Kutchma AJ, Becher A, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-APR-1999) Microbiology, Colorado State University, 
            Fort Collins, CO 80523, USA
REFERENCE   3  (bases 1 to 6755)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6755
                     /organism="Integration vector mini-CTX2"
                     /lab_host="Pseudomonas aeruginosa"
                     /mol_type="other DNA"
                     /db_xref="taxon:106407"
     protein_bind    238..285
                     /label=Flp recombinase binding site
                     /bound_moiety="Flp recombinase"
                     /note="Flp recombinase target site"
     misc_feature    495..521
                     /label=phage CTX attachment site, core sequence
                     /note="phage CTX attachment site, core sequence"
     regulatory      745..769
                     /regulatory_class="terminator"
                     /note="T4 transcription termination signal"
     regulatory      827..846
                     /regulatory_class="promoter"
                     /note="T3 promoter"
     promoter        827..845
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    858..965
                     /label=multiple cloning site
                     /note="multiple cloning site"
     primer_bind     882..898
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(932..948)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      973..992
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     promoter        complement(974..992)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1051..1075
                     /regulatory_class="terminator"
                     /note="T4 transcription termination signal"
     protein_bind    1166..1213
                     /label=Flp recombinase binding site
                     /bound_moiety="Flp recombinase"
                     /note="Flp recombinase target site"
     oriT            1413..1672
     oriT            1420..1528
                     /note="incP origin of transfer"
     CDS             complement(1687..2856)
                     /codon_start=1
                     /gene="int"
                     /product="integrase"
                     /label=int
                     /protein_id="AAF06114.1"
                     /translation="MADGVEVRGKRIRIYFRYQGELCRESIPGDATPENIANAERLAGI
                     INYEIKQGVFSYSRHFPDSPRVKSNTLGHYIDLWLDIKRNQIAASGFRGYTSRVETHIR
                     PRWGDSQADSIDHLDIQDWVQNTLMPKLHNKTVREIVSNLRQIFRLYRTRNRSAHDPTD
                     GIVITLPDADDPDPFTREEIDLILGTETARIGELNLAEFMIWSGPRVSEAIALAWEDVD
                     LDTGTVVFRRARVRSQYKVTKTRRSTRKVQLLAPALRALQQQAKLTRRLPPVQIEVIDR
                     DNRTRKPQRVRFVFHNSASGAAYSTSDTLRNGWWHGHLRNAGVRSRGPNQCRHTFASQM
                     LSSGIATPEWIADQMGHTSTAMIFKHYAKWISKDGPDIVGLLNQALKLS"
     gene            complement(1687..2856)
                     /gene="int"
                     /label=int
                     /note="modified phage CTX integrase gene"
     regulatory      2860..2946
                     /regulatory_class="promoter"
                     /note="trc promoter region with ribosome binding site"
     protein_bind    2877..2893
                     /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        complement(2901..2930)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     CDS             complement(3171..4253)
                     /codon_start=1
                     /gene="lacI"
                     /product="Lac repressor"
                     /label=lacI
                     /protein_id="AAF06115.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            complement(3171..4253)
                     /gene="lacI"
                     /label=lacI
                     /note="GTG start"
     promoter        complement(4254..4331)
                     /gene="lacI"
                     /label=lacI promoter
     misc_feature    4517..4657
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(4843..5431)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5508..6698)
                     /codon_start=1
                     /gene="tet"
                     /product="Tet protein"
                     /label=tet
                     /protein_id="AAF06116.1"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     gene            complement(5508..6698)
                     /gene="tet"
                     /label=tet
                     /note="modified tet gene"
     promoter        complement(6746..19)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"