LOCUS       Exported                8756 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pNG2, complete sequence.
ACCESSION   KR131849
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8756)
  AUTHORS   Gonzalez-Quinonez N, Lopez-Garcia MT, Yague P, Rioseras B, Pisciotta
            A, Alduina R, Manteca A.
  TITLE     New PhiBT1 site-specific integrative vectors with neutral phenotype 
            in Streptomyces
  JOURNAL   Appl. Microbiol. Biotechnol. 100 (6), 2797-2808 (2016)
  PUBMED    26758297
REFERENCE   2  (bases 1 to 8756)
  AUTHORS   Gonzalez-Quinonez N, Lopez-Garcia MT, Yague P, Rioseras B, Manteca 
            A.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-APR-2015) Biologia Funcional, Universidad de Oviedo, 
            c/Julian Claveria s/n Facultad de Medicina, Oviedo 33006, Spain
REFERENCE   3  (bases 1 to 8756)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8756
                     /organism="Cloning vector pNG2"
                     /mol_type="other DNA"
                     /note="derived from Streptomyces coelicolor A3(2) strain 
                     M145"
                     /db_xref="taxon:1780464"
     oriT            2150..2259
                     /note="incP origin of transfer"
     CDS             2292..2663
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     CDS             complement(3373..4371)
                     /codon_start=1
                     /gene="hyg"
                     /product="hygromycin resistance protein"
                     /label=hyg
                     /protein_id="AML61762.1"
                     /translation="MTQESLLLLDRIDSDDSYASLRNDQEFWEPLARRALEELGLPVPP
                     VLRVPGESTNPVLVGEPDPVIKLFGEHWCGPESLASESEAYAVLADAPVPVPRLLGRGE
                     LRPGTGAWPWPYLVMSRMTGTTWRSAMDGTTDRNALLALARELGRVLGRLHRVPLTGNT
                     VLTPHSEVFPELLRERRAATVEDHRGWGYLSPRLLDRLEDWLPDVDTLLAGREPRFVHG
                     DLHGTNIFVDLAATEVTGIVDFTDVYAGDSRYSLVQLHLNAFRGDREILAALLDGAQWK
                     RTEDFARELLAFTFLHDFEVFEETPLDLSGFTDPEELAQFLWGPPDTAPGA"
     gene            complement(3373..4371)
                     /gene="hyg"
                     /label=hyg
     CDS             complement(5056..6366)
                     /codon_start=1
                     /gene="SCO4849"
                     /product="SCO4849 protein"
                     /label=SCO4849
                     /protein_id="AML61763.1"
                     /translation="MVVVFVLVALLMVGVLVTANWYVWRRLFRDTTRAPGPVRRIGAAV
                     IAGGWLLAVGALVAERAGAPFWLQRVLAWPGFLWLALSIYLLLAVLAGEVVRPLLRRFL
                     ERRAAARRTAGAEPSVTPAADTSRIPAGTAPPKTPEAPETPEAPGTEALAAQGSPLAVP
                     SRRLFVSRVVAGAAAAAAVGTVGYGTYGVLSGPKVKRVTVPLAKLPRAAHGFRIAVVSD
                     IHLGPVLGRGFAQQVVDTINSTQPDLIAVVGDLVDGSVKDLGPAAAPLARLTARHGAYF
                     VTGNHEYFSGAEQWVAEVRRLGLLPLENARTELPHFDLAGVNDVAGEDEGQGPDYDRAL
                     GDRDRSRACVLLAHQPVQIHDAVDHGVDLQLSGHTHGGQLWPGNLIAGAANPTLAGLER
                     YGDTQLYVSRGAGAWGPPTRVGAPSDITVIELASRQA"
     gene            complement(5056..6366)
                     /gene="SCO4849"
                     /label=SCO4849
     regulatory      complement(6421..6843)
                     /regulatory_class="promoter"
     rep_origin      complement(7108..7722)
                     /direction=LEFT
     rep_origin      7119..7707
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    7995..8016
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8031..8061
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8069..8085
                     /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)."
     primer_bind     8093..8109
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(8124..8755)
                     /regulatory_class="promoter"
     regulatory      complement(8139..8450)
                     /regulatory_class="terminator"
     terminator      8272..8320
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     regulatory      complement(8631..8634)
                     /regulatory_class="ribosome_binding_site"