LOCUS       Exported                4491 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pJS31, complete sequence.
ACCESSION   KY407916
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4491)
  AUTHORS   Shen J, Chen J, Jensen PR, Solem C.
  TITLE     A novel genetic tool for metabolic optimization of Corynebacterium 
            glutamicum: efficient and repetitive chromosomal integration of 
            synthetic promoter-driven expression libraries
  JOURNAL   Appl. Microbiol. Biotechnol. (2017) In press
  PUBMED    28361238
REFERENCE   2  (bases 1 to 4491)
  AUTHORS   Shen J, Chen J, Jensen PR, Solem C.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-DEC-2016) National Food Institute, Technical 
            University of Denmark, Soltofts Plads Building 221, 056, Lyngby 
            2800, Denmark
REFERENCE   3  (bases 1 to 4491)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4491
                     /organism="Cloning vector pJS31"
                     /mol_type="other DNA"
                     /label=derived from Escherichia coli TOP10
                     /note="derived from Escherichia coli TOP10"
                     /db_xref="taxon:1977027"
     misc_feature    19..464
                     /label=synthetic DNA cassette
                     /note="synthetic DNA cassette"
     misc_feature    19..52
                     /note="lox66; loxP mutant; cre-mediated recombination 
                     occurs in the 8-bp core sequence (GCATACAT)"
     protein_bind    19..52
                     /label=lox66
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      53..100
                     /regulatory_class="terminator"
                     /note="lysA terminator from Corynebacterium glutamicum"
     gene            101..156
                     /gene="attP"
                     /label=attP
     gene            157..199
                     /gene="attB"
                     /label=attB
     misc_feature    200..414
                     /label=multiple cloning site
                     /note="multiple cloning site"
     misc_feature    415..448
                     /note="lox71; loxP mutant; cre-mediated recombination 
                     occurs in the 8-bp core sequence (GCATACAT)"
     protein_bind    415..448
                     /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)."
     oriT            complement(675..1263)
                     /direction=LEFT
                     /note="ori; high-copy-number ColE1/pMB1/pBR322/pUC origin 
                     of replication"
     rep_origin      complement(675..1263)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1439..2860)
                     /codon_start=1
                     /gene="sacB"
                     /product="SacB"
                     /label=sacB
                     /note="negative selection marker; derived from Bacillus 
                     subtilis"
                     /protein_id="ARE67151.1"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVSEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     gene            complement(1439..2860)
                     /gene="sacB"
                     /label=sacB
     misc_feature    complement(1442..2860)
                     /gene="sacB"
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     promoter        complement(2861..3306)
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             complement(3352..4146)
                     /codon_start=1
                     /gene="aph(3)-II (or nptII)"
                     /product="KanR"
                     /label=aph(3)-II (or nptII)
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /protein_id="ARE67152.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(3352..4146)
                     /gene="aph(3)-II (or nptII)"
                     /label=aph(3)-II (or nptII)
     misc_feature    complement(3355..4146)
                     /gene="aph(3)-II (or nptII)"
                     /label=aminoglycoside phosphotransferase from Tn5
                     /note="aminoglycoside phosphotransferase from Tn5"