LOCUS       Exported                7056 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Ascomycete silencing vector pSilent-1, complete sequence.
ACCESSION   LT827033
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7056)
  AUTHORS   Nakayashiki H, Hanada S, Nguyen BQ, Kadotani N, Tosa Y, Mayama S.
  TITLE     RNA silencing as a tool for exploring gene function in ascomycete 
            fungi
  JOURNAL   Fungal genetics and biology : FG & B. 42, 275-283 (2005)
REFERENCE   2  (bases 1 to 7056)
  AUTHORS   De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman 
            Fonseca M, Vanhoucke M, Beyaert R.
  TITLE     BCCM/LMBP Plasmid collection
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 7056)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-MAR-2017) BCCM/LMBP, Universiteit Gent, 
            Technologiepark 927, 9052, BELGIUM
REFERENCE   4  (bases 1 to 7056)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7056
                     /organism="Ascomycete silencing vector pSilent-1"
                     /mol_type="other DNA"
                     /note="BCCM/LMBP Plasmid collection (Ghent University, 
                     Belgium) accession number LMBP 8053."
                     /db_xref="taxon:1969823"
     promoter        24..128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             129..989
                     /codon_start=1
                     /note="unnamed protein product; ampicillin resistance"
                     /protein_id="SLM20106.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1160..1748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      1530..1932
                     /label=pMB1 ori
                     /note="pMB1 ori"
     regulatory      2024..2107
                     /regulatory_class="promoter"
                     /note="lac promoter"
     protein_bind    2036..2057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2072..2102
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     misc_feature    2108..2145
                     /label=5 UTR of lacZ, incl. RBS
                     /note="5 UTR of lacZ, incl. RBS"
     protein_bind    2110..2126
                     /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     2134..2150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2171..2189
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      2171..2187
                     /regulatory_class="promoter"
                     /note="T3 promoter"
     regulatory      2235..2592
                     /regulatory_class="promoter"
                     /note="trpC promoter"
     CDS             2593..3618
                     /codon_start=1
                     /note="unnamed protein product; hygromycin resistance"
                     /protein_id="SLM20107.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     regulatory      3638..4123
                     /regulatory_class="terminator"
                     /note="trpC terminator"
     primer_bind     4132..4148
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      4162..5447
                     /regulatory_class="promoter"
                     /note="trpC promoter"
     primer_bind     complement(5444..5460)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     intron          5491..5624
                     /note="CUT intron"
     regulatory      5666..6398
                     /regulatory_class="terminator"
                     /note="trpC terminator"
     promoter        complement(6413..6431)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      complement(6415..6431)
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     primer_bind     complement(6441..6457)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6599..7054
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 ori"