LOCUS       Exported                5709 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pNB20, complete sequence.
ACCESSION   KT826597
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5709)
  AUTHORS   Bongio NJ, Lampe DJ.
  TITLE     Inhibition of Plasmodium berghei Development in Mosquitoes by 
            Effector Proteins Secreted from Asaia sp. Bacteria Using a Novel 
            Native Secretion Signal
  JOURNAL   PLoS ONE 10 (12), E0143541 (2015)
  PUBMED    26636338
REFERENCE   2  (bases 1 to 5709)
  AUTHORS   Bongio NJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-SEP-2015) Biology, Shenandoah University, 1460 
            University Drive, Winchester, VA 22601, USA
REFERENCE   3  (bases 1 to 5709)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5709
                     /organism="Cloning vector pNB20"
                     /mol_type="other DNA"
                     /db_xref="taxon:1766770"
     CDS             458..1252
                     /codon_start=1
                     /gene="neo"
                     /product="neomycin phosphotransferase"
                     /label=neo
                     /protein_id="ALR84549.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            458..1252
                     /gene="neo"
                     /label=neo
     protein_bind    1657..1678
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1693..1723
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1731..1747
                     /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)."
     regulatory      1767..1783
                     /regulatory_class="enhancer"
                     /note="translational enhancer"
     CDS             1791..2651
                     /codon_start=1
                     /product="beta lactamase"
                     /label=beta lactamase
                     /protein_id="ALR84548.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3255..3917
     CDS             complement(3255..3917)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      3918..4687
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"