LOCUS       Exported                9649 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector phis-5GFP, complete sequence.
ACCESSION   JX156632
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9649)
  AUTHORS   Bowring FJ, Yeadon PJ, Catcheside DE.
  TITLE     Use of fluorescent protein to analyse recombination at three loci in
            Neurospora crassa
  JOURNAL   Fungal Genet. Biol. 49 (8), 619-625 (2012)
  PUBMED    22691725
REFERENCE   2  (bases 1 to 9649)
  AUTHORS   Bowring FJ, Yeadon J, Catcheside DEA.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-JUN-2012) School of Biological Sciences, Flinders 
            University, Sturt Road, Bedford Park, SA 5162, Australia
REFERENCE   3  (bases 1 to 9649)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9649
                     /organism="Cloning vector phis-5GFP"
                     /mol_type="other DNA"
                     /db_xref="taxon:1198189"
     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 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        239..257
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     misc_feature    336..1578
                     /function="targeting to the his-5 locus"
                     /label=truncated his-5 gene
                     /note="truncated his-5 gene"
     regulatory      1640..2561
                     /regulatory_class="promoter"
                     /gene="ccg-1"
                     /note="Neurospora crassa ccg-1 (grg-1) promoter"
     gene            1640..2561
                     /gene="ccg-1"
                     /label=ccg-1
     primer_bind     2562..2578
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(2583..2705,2760..2786,2967..2989,3155..3285,3353..3755,
                     3765..4485)
                     /codon_start=1
                     /gene="h1/sgfp fusion"
                     /product="histone h1/green fluorescent protein S65T variant
                     fusion protein"
                     /label=h1/sgfp fusion
                     /protein_id="AFP67894.1"
                     /translation="MPPKKTETKAADASAAAAPAPAAAPTSAPKTKSPSTHASYLDMIT
                     DAIVALKDRAGSSRQALKKYVRANNTLGNVTDNMFDSLFNKALKNGVDKGVFEQPKGPS
                     GGTKLAKKVAKPAPKKAAPKKETKEKKPAAAKKEGAAKKETKEKKAPAAKKAAAPKKAA
                     APKKEVKEKKAAAPKKKAAAPAVADKETVLTKTKSGRVAKSTAKPAAAKKAAAPKKAAA
                     SKKAEKAEPAAEKAMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     gene            2583..4485
                     /gene="h1/sgfp fusion"
                     /label=h1/sgfp fusion
     CDS             3766..4485
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    4494..6013
                     /function="targeting to the his-5 locus"
                     /label=3 flank ofNeurospora crassa his-5 gene
                     /note="3 flank ofNeurospora crassa his-5 gene"
     promoter        complement(6083..6101)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6108..6124)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6265..6693
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             7037..7831
                     /codon_start=1
                     /gene="neoR"
                     /product="neomycin resistance protein"
                     /label=neoR
                     /protein_id="AFP67892.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            7037..7831
                     /gene="neoR"
                     /label=neoR
     CDS             7849..8709
                     /codon_start=1
                     /gene="bla"
                     /product="ampicillin resistance protein"
                     /label=bla
                     /protein_id="AFP67893.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            7849..8709
                     /gene="bla"
                     /label=bla
     rep_origin      8880..9468
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"