LOCUS       Exported                9650 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector phis-3GFP3, complete sequence.
ACCESSION   JX156631
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9650)
  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 9650)
  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 9650)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9650
                     /organism="Cloning vector phis-3GFP3"
                     /mol_type="other DNA"
                     /db_xref="taxon:1198188"
     promoter        363..467
                     /gene="bla"
                     /label=AmpR promoter
     CDS             468..1328
                     /codon_start=1
                     /gene="bla"
                     /product="beta lactamase"
                     /function="ampicillin resistance protein"
                     /label=bla
                     /protein_id="AFP67891.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            468..1328
                     /gene="bla"
                     /label=bla
     rep_origin      1499..2087
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        2345..2363
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(2374..4006)
                     /function="targeting to the his-3 locus"
                     /label=3 flank of Neurospora crassa his-3 gene
                     /note="3 flank of Neurospora crassa his-3 gene"
     CDS             2638..2649
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        4008..4026
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4059..4980
                     /regulatory_class="promoter"
                     /gene="ccg-1"
                     /note="Neurospora crassa ccg-1 (grg-1) promoter"
     gene            4059..4980
                     /gene="ccg-1"
                     /label=ccg-1
     primer_bind     4981..4997
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    5002..6904
                     /function="inactive h1/sgfp fusion as substrate for meiotic
                     recombination"
                     /label=3 allele at 6815 renders sgfp non-functional
                     /note="3 allele at 6815 renders sgfp non-functional"
     CDS             6185..6904
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALS*DPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6925..6941)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(6971..6989)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    complement(7532..9633)
                     /function="targeting to the his-3 locus"
                     /label=truncated his-3 gene
                     /note="truncated his-3 gene"