LOCUS       Exported               10400 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pMF309, complete sequence.
ACCESSION   AY598430
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10400)
  AUTHORS   Freitag M, Hickey PC, Raju NB, Selker EU, Read ND.
  TITLE     GFP as a tool to analyze the organization, dynamics and function of 
            nuclei and microtubules in Neurospora crassa
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10400)
  AUTHORS   Freitag M, Selker EU.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-APR-2004) Inst. of Mol. Biology, University of Oregon,
            OR, USA
REFERENCE   3  (bases 1 to 10400)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10400
                     /organism="Expression vector pMF309"
                     /mol_type="other DNA"
                     /db_xref="taxon:274627"
     regulatory      complement(1..17)
                     /regulatory_class="promoter"
                     /note="SP6 promoter"
     promoter        363..467
                     /gene="bla"
                     /label=AmpR promoter
     CDS             468..1328
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /function="ampicillin resistance"
                     /label=bla
                     /protein_id="AAT34985.1"
                     /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"
     regulatory      complement(2345..2365)
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     promoter        2345..2363
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(2374..4006)
                     /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"
     misc_feature    complement(4008..4058)
                     /note="T7 promoter and multiple cloning site from 
                     pBluescript KS+"
     promoter        4008..4026
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4059..4980
                     /regulatory_class="promoter"
                     /note="Neurospora crassa ccg-1 (grg-1) promoter"
     primer_bind     4981..4997
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(5007..5018,5259..5282,5357..5382,5451..5492,5558..5612,
                     5686..6476,6534..7653)
                     /codon_start=1
                     /gene="bml/sgfp fusion"
                     /product="beta-tubulin/green fluorescent protein S65T 
                     variant fusion protein"
                     /label=bml/sgfp fusion
                     /note="Bml/GFP"
                     /protein_id="AAT34986.1"
                     /translation="MREIVHLQTGQCGNQIGAAFWQTISGEHGLDASGVYNGTSELQLE
                     RMNVYFNEASGNKYVPRAVLVDLEPGTMDAVRAGPFGQLFRPDNFVFGQSGAGNNWAKG
                     HYTEGAELVDQVLDVVRREAEGCDCLQGFQITHSLGGGTGAGMGTLLISKIREEFPDRM
                     MATYSVVPSPKVSDTVVEPYNATLSVHQLVENSDETFCIDNEALYDICMRTLKLSNPSY
                     GDLNHLVSAVMSGVTVSLRFPGQLNSDLRKLAVNMVPFPRLHFFMVGFAPLTSRGAHHF
                     RAVSVPELTQQMFDPKNMMAASDFRNGRYLTCSAIFRGKVSMKEVEDQMRNVQNKNSSY
                     FVEWIPNNVQTALCSIPPRGLKMSSTFVGNSTAIQELFKRIGEQFTAMFKRKAFLHWYT
                     GEGMDEMEFTEAESNMNDLVSEYQQYQDAGVDEEEEEYEEEAPLEGEELINMVSKGEEL
                     FTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFTY
                     GVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIEL
                     KGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQ
                     NTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     gene            5007..7653
                     /gene="bml/sgfp fusion"
                     /label=bml/sgfp fusion
     CDS             6934..7653
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    complement(7654..7738)
                     /note="T3 promoter and multiple cloning site from 
                     pBluescript KS+"
     primer_bind     complement(7674..7690)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(7720..7738)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    complement(8280..10381)
                     /note="5 truncated histidine biosynthesis trifunctional 
                     protein; includes phosphoribosyl-AMP cyclohydrolase, 
                     phosphoribosyl-ATP cyclohydrolase and histindinol 
                     dehydrogenase"
     misc_feature    complement(10382..10397)
                     /label=multiple cloning site from pSP72
                     /note="multiple cloning site from pSP72"
     promoter        complement(10399..17)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"