LOCUS       Exported                9482 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Inducible shuttle vector pPW578, complete sequence.
ACCESSION   DQ164216
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9482)
  AUTHORS   Woo PC, Ma SS, Teng JL, Li MW, Kao RY, Lau SK, Yuen KY.
  TITLE     Construction of an inducible expression shuttle vector for 
            Laribacter hongkongensis, a novel bacterium associated with 
            gastroenteritis
  JOURNAL   FEMS Microbiol. Lett. 252 (1), 57-65 (2005)
  PUBMED    16165318
REFERENCE   2  (bases 1 to 9482)
  AUTHORS   Woo PCY., Ma SSL., Teng JLL., Li MWS., Lau SKP., Yuen KY.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-AUG-2005) Microbiology, The University of Hong Kong, 
            Hong Kong
REFERENCE   3  (bases 1 to 9482)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9482
                     /organism="Inducible shuttle vector pPW578"
                     /lab_host="Laribacter hongkongensis HLHK8"
                     /mol_type="other DNA"
                     /db_xref="taxon:342568"
     rep_origin      7..462
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     polyA_signal    469..590
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     952..968
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        975..993
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1981..2604
                     /codon_start=1
                     /gene="parA"
                     /product="ATPase"
                     /label=parA
                     /note="involved in chromosome partitioning"
                     /protein_id="AAZ91681.1"
                     /translation="MRTIVVASQKGGVGKTTIAGHLGVMAEQSKEGPVALIDTDPQGSL
                     ASWWNERTDEAPLFARVEINKLTEHLQALAKGGIKLAIIDTPPSVTEMIQQVLRTADLV
                     LIPTRPSPHDLRAVGSTVELVENAGKRMIFVINGAAPRARIAGEAAVALSQHGTVAPVT
                     LYQRTDFASSMIDGRTVQEIDPKGRSAEEIGKLWKYVSTQLRKI"
     gene            1981..2604
                     /gene="parA"
                     /label=parA
     CDS             3058..3975
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein"
                     /label=repA
                     /note="plasmid replication initiator protein"
                     /protein_id="AAZ91682.1"
                     /translation="MAEVVDEQPARRKRTPKGPRGLMPVHHPNRDFFLCDLFNYALKDD
                     GVSMEAPIFTLATKPDTSVWHWESKDGARAITVTPSVKGRATQFDKDLLIYVVSQMTEA
                     MNRGRPDANSRTVRFRVYDYLVSTNKPTGGKEYQRLEDALDRLRGTSIKTNIKTGGQRV
                     KEGFGIVDSWTIIEKAPDDDRMIAVEVTLSKWLFNAVQAHEVLTINPDYFRLRKPIERR
                     LYELARKHCGDQAFFVIGLELLQDKCGSKSALFEFRRALREIIKADTLPDYRMTLDNEK
                     DQVIFYTRDAKKLTASAALPQRFQ"
     gene            3058..3975
                     /gene="repA"
                     /label=repA
     misc_feature    5829..5866
                     /label=multiple cloning site
                     /note="multiple cloning site"
     regulatory      5867..6026
                     /regulatory_class="promoter"
                     /note="tetracycline inducible promoter region"
     protein_bind    5899..5917
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    5984..6002
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     misc_feature    6027..6650
                     /label=tetracycline repressor region
                     /note="tetracycline repressor region"
     CDS             6027..6650
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     rep_origin      complement(6974..7562)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     polyA_signal    7891..7938
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             complement(8170..8964)
                     /codon_start=1
                     /gene="aph(3)-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(8999..9356)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9013..9148
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(9358..9462)
                     /gene="bla"
                     /label=AmpR promoter