LOCUS       Exported                7072 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pDEST375, complete sequence.
ACCESSION   KC614689
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7072)
  AUTHORS   Giuraniuc CV, Macpherson M, Saka Y.
  TITLE     Gateway Vectors for Efficient Artificial Gene Assembly In Vitro and 
            Expression in Yeast Saccharomyces cerevisiae
  JOURNAL   PLoS ONE 8 (5), E64419 (2013)
  PUBMED    23675537
REFERENCE   2  (bases 1 to 7072)
  AUTHORS   Giuraniuc C, Macpherson M, Saka Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-FEB-2013) School of Medical Sciences, Institute of 
            Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen 
            AB25 2ZD, UK
REFERENCE   3  (bases 1 to 7072)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7072
                     /organism="Cloning vector pDEST375"
                     /mol_type="other DNA"
                     /db_xref="taxon:1333695"
     misc_feature    1..4879
                     /label=pIS375 vector backbone
                     /note="pIS375 vector backbone"
     misc_feature    complement(1..784)
                     /label=5 flanking MET15 YIp-Out fragment
                     /note="5 flanking MET15 YIp-Out fragment"
     misc_feature    787..4261
                     /label=YIplac211 vector backbone
                     /note="YIplac211 vector backbone"
     gene            complement(1105..2213)
                     /gene="ura3"
                     /label=ura3
     CDS             complement(1184..1987)
                     /codon_start=1
                     /gene="ura3"
                     /product="orotidine-5-phosphate decarboxylase"
                     /label=ura3
                     /note="OMP decarboxylase"
                     /protein_id="AGL76312.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     promoter        1990..2204
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     promoter        2301..2405
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2406..3266
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="AmpR"
                     /protein_id="AGL76313.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            2406..3266
                     /gene="bla"
                     /label=bla
     rep_origin      3421..4040
                     /label=pBR322 origin of replication
                     /note="pBR322 origin of replication"
     rep_origin      3437..4025
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4265..4866)
                     /label=3 flanking MET15 YIp-In fragment
                     /note="3 flanking MET15 YIp-In fragment"
     regulatory      complement(4920..5122)
                     /regulatory_class="terminator"
                     /gene="tef"
                     /note="Ashbya gossypii TEF terminator"
     gene            complement(4920..5122)
                     /gene="tef"
                     /label=tef
     terminator      4920..5117
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     misc_recomb     5143..5267
                     /label=attR1 recombination site
                     /note="attR1 recombination site"
     protein_bind    5143..5267
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        5292..5322
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             5376..6035
                     /codon_start=1
                     /gene="cmr"
                     /product="chloramphenicol resistance protein"
                     /label=cmr
                     /protein_id="AGL76314.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            5376..6035
                     /gene="cmr"
                     /label=cmr
     CDS             6377..6682
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB protein"
                     /label=ccdB
                     /protein_id="AGL76315.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            6377..6682
                     /gene="ccdB"
                     /label=ccdB
     misc_recomb     complement(6723..6847)
                     /label=attR2 recombination site
                     /note="attR2 recombination site"
     protein_bind    complement(6723..6847)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      6858..6986
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     primer_bind     complement(6897..6916)
                     /label=T7 reverse primer
                     /note="T7 reverse primer"
     terminator      6914..6961
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"