LOCUS       Exported                3430 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pRI857, complete sequence.
ACCESSION   KX774470
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3430)
  AUTHORS   Zhang K, Su H, Yang M, Ge J, Li G, Yi J, Wang Y.
  TITLE     Construction of a thermo-sensitive pRI857 vector for efficient DNA 
            capturing in Escherichia coli
  JOURNAL   Biotechnol. Lett. (2017) In press
  PUBMED    28251389
REFERENCE   2  (bases 1 to 3430)
  AUTHORS   Zhang K, Wang Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-AUG-2016) College of Environmental and Biological 
            Engineering, Nanjing University of Science and Technology, 200 
            Xiaolingwei, Nanjing, Jiangsu 210094, China
REFERENCE   3  (bases 1 to 3430)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3430
                     /organism="Cloning vector pRI857"
                     /mol_type="other DNA"
                     /db_xref="taxon:1965237"
     gene            complement(1..837)
                     /gene="KanR"
                     /label=KanR
     CDS             complement(1..795)
                     /codon_start=1
                     /gene="KanR"
                     /product="KanR"
                     /label=KanR
                     /note="kanamycin resistance determinant"
                     /protein_id="AQX17852.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     regulatory      complement(797..837)
                     /regulatory_class="promoter"
                     /gene="KanR"
     RBS             799..807
                     /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)"
     protein_bind    950..971
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        986..1016
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1024..1040
                     /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)."
     CDS             1060..1773
                     /codon_start=1
                     /gene="cI857"
                     /product="cI857"
                     /label=cI857
                     /note="cI repressor"
                     /protein_id="AQX17851.1"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLTKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPKLRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     gene            1060..1773
                     /gene="cI857"
                     /label=cI857
     regulatory      2026..2062
                     /regulatory_class="terminator"
     primer_bind     complement(2117..2133)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2162..2180)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    2204..2237
                     /label=lox71
                     /bound_moiety="Cre recombinase"
                     /note="Left element (LE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    2259..2275
                     /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(2283..2313)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2328..2349
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2637..3225)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"