LOCUS       Exported                8875 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Integration vector mini-CTX-lacZ, complete sequence.
ACCESSION   AF140579
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8875)
  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  JOURNAL   Unpublished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 8875)
  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 8875)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8875
                     /organism="Integration vector mini-CTX-lacZ"
                     /lab_host="Pseudomonas aeruginosa"
                     /mol_type="other DNA"
                     /db_xref="taxon:106409"
     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"
     CDS             complement(921..3995)
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /protein_id="AAF06120.1"
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYG
                     QDSRLPSEFDLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQ
                     ISDFHVATRFNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIID
                     ERGGYADRVTLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIEN
                     GLLLLNGKPLLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLW
                     YTLCDRYGLYVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWS
                     LGNESGHGANHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVP
                     KWSIKKWLSLPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSL
                     IKYDENGNPWSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQ
                     TIEVTSEYLFRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQ
                     LWLTVRVVQPNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELG
                     NKRWQFNRQSGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWK
                     AAGHYQAEAALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDV
                     EVASDTPHPARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPY
                     VFPSENGLRCGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDG
                     FHMGIGGDDSWSPSVSAEFQLSAGRYHYQLVWCQK"
     gene            complement(921..3995)
                     /gene="lacZ"
                     /label=lacZ
     misc_feature    972..977
                     /note="location of uncharacterized mutation which removes 
                     EcoRI site"
     primer_bind     3959..3975
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3991..4007)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    4059..4135
                     /label=RNAse III processing site
                     /note="RNAse III processing site"
     misc_feature    4159..4230
                     /label=multiple cloning site
                     /note="multiple cloning site"
     primer_bind     complement(4197..4213)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      4238..4257
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     promoter        complement(4239..4257)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4316..4340
                     /regulatory_class="terminator"
                     /note="T4 transcription termination signal"
     protein_bind    4431..4478
                     /label=Flp recombinase binding site
                     /bound_moiety="Flp recombinase"
                     /note="Flp recombinase target site"
     oriT            4678..4937
     oriT            4685..4793
                     /note="incP origin of transfer"
     CDS             complement(4952..6121)
                     /codon_start=1
                     /gene="int"
                     /product="integrase"
                     /label=int
                     /protein_id="AAF06121.1"
                     /translation="MADGVEVRGKRIRIYFRYQGELCRESIPGDATPENIANAERLAGI
                     INYEIKQGVFSYSRHFPDSPRVKSNTLGHYIDLWLDIKRNQIAASGFRGYTSRVETHIR
                     PRWGDSQADSIDHLDIQDWVQNTLMPKLHNKTVREIVSNLRQIFRLYRTRNRSAHDPTD
                     GIVITLPDADDPDPFTREEIDLILGTETARIGELNLAEFMIWSGPRVSEAIALAWEDVD
                     LDTGTVVFRRARVRSQYKVTKTRRSTRKVQLLAPALRALQQQAKLTRRLPPVQIEVIDR
                     DNRTRKPQRVRFVFHNSASGAAYSTSDTLRNGWWHGHLRNAGVRSRGPNQCRHTFASQM
                     LSSGIATPEWIADQMGHTSTAMIFKHYAKWISKDGPDIVGLLNQALKLS"
     gene            complement(4952..6121)
                     /gene="int"
                     /label=int
                     /note="modified phage CTX integrase gene"
     rep_origin      complement(6218..6806)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7628..8818)
                     /codon_start=1
                     /gene="tet"
                     /product="Tet protein"
                     /label=tet
                     /protein_id="AAF06122.1"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     gene            complement(7628..8818)
                     /gene="tet"
                     /label=tet
                     /note="modified tet gene"
     promoter        complement(8866..19)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"