LOCUS       Exported                3581 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPh079, complete sequence.
ACCESSION   KT003680
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3581)
  AUTHORS   Krom RJ, Bhargava P, Lobritz MA, Collins JJ.
  TITLE     Engineered Phagemids for Nonlytic, Targeted Antibacterial Therapies
  JOURNAL   Nano Lett. 15 (7), 4808-4813 (2015)
  PUBMED    26044909
REFERENCE   2  (bases 1 to 3581)
  AUTHORS   Krom RJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-JUN-2015) Molecular Medicine, Boston University, 72 
            East Concord St, Boston, MA 02118, USA
REFERENCE   3  (bases 1 to 3581)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3581
                     /organism="Cloning vector pPh079"
                     /mol_type="other DNA"
                     /db_xref="taxon:1680897"
     regulatory      31..104
                     /regulatory_class="promoter"
                     /note="PLtet0"
     promoter        31..104
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    31..49
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    56..74
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             142..264
                     /codon_start=1
                     /product="cecropin PR-39"
                     /label=cecropin PR-39
                     /protein_id="AKQ98650.1"
                     /translation="MRRRPRPPYLPRPRPPPFFPPRLPPRIPPGFPPRFPPRFP"
     CDS             313..435
                     /codon_start=1
                     /product="cecropin PR-39"
                     /label=cecropin PR-39
                     /protein_id="AKQ98651.1"
                     /translation="MRRRPRPPYLPRPRPPPFFPPRLPPRIPPGFPPRFPPRFP"
     regulatory      442..564
                     /regulatory_class="terminator"
                     /note="T0"
     terminator      456..550
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     regulatory      631..704
                     /regulatory_class="promoter"
                     /note="PLtet0"
     promoter        631..704
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    631..649
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    656..674
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             730..738
                     /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             745..804
                     /codon_start=1
                     /product="apidaecin"
                     /label=apidaecin
                     /protein_id="AKQ98652.1"
                     /translation="MGNNRPVYIPQPRPPHPRI"
     RBS             843..851
                     /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             857..916
                     /codon_start=1
                     /product="apidaecin"
                     /label=apidaecin
                     /protein_id="AKQ98653.1"
                     /translation="MGNNRPVYIPQPRPPHPRI"
     regulatory      923..1045
                     /regulatory_class="terminator"
                     /note="T0"
     terminator      937..1031
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      1148..1576
                     /label=F1
                     /note="F1"
     terminator      1600..1686
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(1850..2438)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      2526..2620
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2651..3445)
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"