LOCUS       Exported                9225 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pNG3, complete sequence.
ACCESSION   KR131850
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9225)
  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 9225)
  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 9225)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9225
                     /organism="Cloning vector pNG3"
                     /mol_type="other DNA"
                     /note="derived from Streptomyces coelicolor A3(2) strain 
                     M145"
                     /db_xref="taxon:1780465"
     rep_origin      complement(254..868)
                     /direction=LEFT
     rep_origin      265..853
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    1141..1162
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1177..1207
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1215..1231
                     /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     1239..1255
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     oriT            3480..3589
                     /note="incP origin of transfer"
     CDS             3622..3993
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     CDS             complement(4703..5701)
                     /codon_start=1
                     /gene="hyg"
                     /product="hygromycin resistance protein"
                     /label=hyg
                     /protein_id="AML61764.1"
                     /translation="MTQESLLLLDRIDSDDSYASLRNDQEFWEPLARRALEELGLPVPP
                     VLRVPGESTNPVLVGEPDPVIKLFGEHWCGPESLASESEAYAVLADAPVPVPRLLGRGE
                     LRPGTGAWPWPYLVMSRMTGTTWRSAMDGTTDRNALLALARELGRVLGRLHRVPLTGNT
                     VLTPHSEVFPELLRERRAATVEDHRGWGYLSPRLLDRLEDWLPDVDTLLAGREPRFVHG
                     DLHGTNIFVDLAATEVTGIVDFTDVYAGDSRYSLVQLHLNAFRGDREILAALLDGAQWK
                     RTEDFARELLAFTFLHDFEVFEETPLDLSGFTDPEELAQFLWGPPDTAPGA"
     gene            complement(4703..5701)
                     /gene="hyg"
                     /label=hyg
     CDS             complement(6386..7696)
                     /codon_start=1
                     /gene="SCO4849"
                     /product="SCO4849 protein"
                     /label=SCO4849
                     /protein_id="AML61765.1"
                     /translation="MVVVFVLVALLMVGVLVTANWYVWRRLFRDTTRAPGPVRRIGAAV
                     IAGGWLLAVGALVAERAGAPFWLQRVLAWPGFLWLALSIYLLLAVLAGEVVRPLLRRFL
                     ERRAAARRTAGAEPSVTPAADTSRIPAGTAPPKTPEAPETPEAPGTEALAAQGSPLAVP
                     SRRLFVSRVVAGAAAAAAVGTVGYGTYGVLSGPKVKRVTVPLAKLPRAAHGFRIAVVSD
                     IHLGPVLGRGFAQQVVDTINSTQPDLIAVVGDLVDGSVKDLGPAAAPLARLTARHGAYF
                     VTGNHEYFSGAEQWVAEVRRLGLLPLENARTELPHFDLAGVNDVAGEDEGQGPDYDRAL
                     GDRDRSRACVLLAHQPVQIHDAVDHGVDLQLSGHTHGGQLWPGNLIAGAANPTLAGLER
                     YGDTQLYVSRGAGAWGPPTRVGAPSDITVIELASRQA"
     gene            complement(6386..7696)
                     /gene="SCO4849"
                     /label=SCO4849
     regulatory      complement(7757..8179)
                     /regulatory_class="promoter"
     CDS             complement(8297..9157)
                     /codon_start=1
                     /gene="Bla"
                     /product="ampicillin resistance protein"
                     /label=Bla
                     /protein_id="AML61766.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(8297..9157)
                     /gene="Bla"
                     /label=Bla