LOCUS       Exported                6695 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Neurospora cloning vector pTH1067.9, complete sequence.
ACCESSION   JF749198
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6695)
  AUTHORS   Hammond TM, Xiao H, Rehard DG, Boone EC, Perdue TD, Pukkila PJ, Shiu
            PK.
  TITLE     Fluorescent and bimolecular-fluorescent protein tagging of genes at 
            their native loci in Neurospora crassa using specialized 
            double-joint PCR plasmids
  JOURNAL   Fungal Genet. Biol. 48 (9), 866-873 (2011)
  PUBMED    21664475
REFERENCE   2  (bases 1 to 6695)
  AUTHORS   Hammond TM, Shiu PKT.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-MAR-2011) Biological Sciences, University of 
            Missouri-Columbia, 103 Tucker Hall, Columbia, MO 65211, USA
REFERENCE   3  (bases 1 to 6695)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6695
                     /organism="Neurospora cloning vector pTH1067.9"
                     /mol_type="other DNA"
                     /db_xref="taxon:1051833"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     205..221
                     /label=M13 reverse primer
                     /note="M13 reverse primer"
     promoter        239..257
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     239..256
                     /label=SP6 primer
                     /note="SP6 primer"
     regulatory      337..700
                     /regulatory_class="promoter"
                     /note="trpC"
     CDS             701..1726
                     /codon_start=1
                     /product="hph"
                     /label=hph
                     /note="hygromycin resistance protein"
                     /protein_id="AEJ38146.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANILFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     regulatory      complement(1755..2314)
                     /regulatory_class="terminator"
                     /note="trpC"
     CDS             complement(2315..3034)
                     /codon_start=1
                     /product="GFP"
                     /label=GFP
                     /protein_id="AEJ38145.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    complement(3035..3058)
                     /label=4X(GlyAla)-linker
                     /note="4X(GlyAla)-linker"
     primer_bind     complement(3128..3147)
                     /label=T7 primer
                     /note="T7 primer"
     promoter        complement(3129..3147)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3154..3170)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3155..3170)
                     /label=M13 (-20) forward primer
                     /note="M13 (-20) forward primer"
     rep_origin      3311..3739
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      3312..3749
                     /label=f1
                     /note="f1"
     CDS             4083..4877
                     /codon_start=1
                     /product="Kan(R)"
                     /label=Kan
                     /note="kanamycin resistance protein"
                     /protein_id="AEJ38143.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4895..5755
                     /codon_start=1
                     /product="Amp(R)"
                     /label=Amp
                     /note="ampicillin resistance protein"
                     /protein_id="AEJ38144.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5900..6573
                     /label=pUC
                     /note="pUC"
     rep_origin      5926..6514
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"