LOCUS       Exported                7720 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Triparental mating vector pCD13SK-Flp-oriT, complete sequence.
ACCESSION   EU034638
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7720)
  AUTHORS   Son MS, Nguyen DT, Kang Y, Hoang TT.
  TITLE     Engineering of FRT-lacZ fusion constructs: induction of the 
            Pseudomonas aeruginosa fadAB1 operon by medium and long chain-length
            fatty acids
  JOURNAL   Plasmid 59 (2), 111-118 (2008)
  PUBMED    18221997
REFERENCE   2  (bases 1 to 7720)
  AUTHORS   Son MS, Kang Y, Nguyen DT, Hoang TT.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-JUL-2007) Microbiology, University of Hawaii Manoa, 
            2538 McCarthy Mall, Honolulu, HI 96822, USA
REFERENCE   3  (bases 1 to 7720)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7720
                     /organism="Triparental mating vector pCD13SK-Flp-oriT"
                     /mol_type="other DNA"
                     /db_xref="taxon:475612"
     protein_bind    678..909
                     /gene="attP"
                     /label=-lambda attP
                     /bound_moiety="lambda integrase"
                     /note="integrase from phage lambda"
     CDS             complement(1798..2652)
                     /codon_start=1
                     /gene="aad"
                     /product="aminoglycoside adenylyltransferase"
                     /label=aad
                     /note="streptomycin/spectinomycin resistance"
                     /protein_id="ABW06018.1"
                     /translation="MLWSSNDVTQQGSRPKTKLNIMREAVIAEVSTQLSEVVGVIERHL
                     EPTLLAVHLYGSAVDGGLKPHSDIDLLVTVTVRLDETTRRALINDLLETSASPGESEIL
                     RAVEVTIVVHDDIIPWRYPAKRELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHS
                     VALVGPAAEELFDPVPEQDLFEALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGK
                     IAPKDVAADWAMERLPAQYQPVILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKV
                     VGK"
     gene            complement(1798..2652)
                     /gene="aad"
                     /label=aad
     CDS             complement(1798..2589)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      complement(3413..3841)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     3985..4001
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4011..4029
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     4055..4071
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     oriT            4636..4744
                     /note="incP origin of transfer"
     CDS             complement(4958..6229)
                     /codon_start=1
                     /gene="flp"
                     /product="Flp"
                     /label=flp
                     /note="flippase; recognizes and recombines flp recognition 
                     targets (FRT)"
                     /protein_id="ABW06019.1"
                     /translation="MPQFGILCKTPPKVLVRQFVERFERPSGEKIALCAAELTYLCWMI
                     THNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWEFTI
                     IPYYGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEKILN
                     SFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCLVTET
                     KTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLKDNLVR
                     SYNKALKKNAPYSIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKRASAVAR
                     TTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKGSAEGSIR
                     YPAWNGIISQEVLDYLSSYINRRI"
     gene            complement(4958..6229)
                     /gene="flp"
                     /label=flp
     RBS             6233..6241
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3 end of the 16S rRNA (Chen et al., 1994)"
     CDS             6330..7043
                     /codon_start=1
                     /gene="cI857"
                     /product="cI857"
                     /label=cI857
                     /note="temperature sensitive lambda-repressor"
                     /protein_id="ABW06020.1"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLTKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPKLRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     gene            6330..7043
                     /gene="cI857"
                     /label=cI857
     promoter        complement(7329..7347)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7368..7384)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7392..7408
                     /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(7416..7446)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7461..7482
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."