LOCUS       Exported                8047 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pDEST415TEFt7, complete sequence.
ACCESSION   JX901381
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8047)
  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 8047)
  AUTHORS   Giuraniuc C, Saka Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-OCT-2012) School of Medical Sciences, Institute of 
            Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen 
            AB25 2ZD, UK
REFERENCE   3  (bases 1 to 8047)
  AUTHORS   Giuraniuc C, Saka Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-JUL-2015) School of Medical Sciences, Institute of 
            Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen 
            AB25 2ZD, UK
REFERENCE   4  (bases 1 to 8047)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     On Jul 28, 2015 this sequence version replaced JX901381.1.
FEATURES             Location/Qualifiers
     source          1..8047
                     /organism="Cloning vector pDEST415TEFt7"
                     /mol_type="other DNA"
                     /db_xref="taxon:1333773"
     CDS             complement(663..1769)
                     /codon_start=1
                     /gene="LEU2"
                     /product="LEU2"
                     /label=LEU2
                     /note="derived from S. cerevisiae LEU2"
                     /protein_id="AGL80163.1"
                     /translation="MSAPMSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDF
                     ENHLIGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKEL
                     QLYANLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDS
                     EQYTVPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLK
                     VQHQLIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDK
                     NTAFGLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAG
                     IRTGDLGGSNSTTEVGDAVAEEVKKILA"
     gene            complement(663..1769)
                     /gene="LEU2"
                     /label=LEU2
     CDS             complement(663..1757)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(1770..2174)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(2474..2929)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      complement(2477..2932)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 ori"
     primer_bind     3074..3090
                     /label=M13 -20 primer
                     /note="M13 -20 primer"
     regulatory      3100..3119
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     promoter        3100..3118
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      complement(3147..3275)
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     terminator      3172..3219
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3217..3236
                     /label=T7 reverse primer
                     /note="T7 reverse primer"
     misc_recomb     3286..3410
                     /label=attR2 recombination site
                     /note="attR2 recombination site"
     protein_bind    3286..3410
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(3451..3756)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB"
                     /label=ccdB
                     /protein_id="AGL80164.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            complement(3451..3756)
                     /gene="ccdB"
                     /label=ccdB
     CDS             complement(4098..4757)
                     /codon_start=1
                     /gene="CmR"
                     /product="chloramphenicol resistance protein"
                     /label=CmR
                     /protein_id="AGL80165.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(4098..4757)
                     /gene="CmR"
                     /label=CmR
     promoter        complement(4811..4841)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     misc_recomb     complement(4866..4990)
                     /label=attR1 recombination site
                     /note="attR1 recombination site"
     protein_bind    complement(4866..4990)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      5011..5213
                     /regulatory_class="terminator"
                     /gene="TEF"
                     /note="derived from A. gossypii TEF terminator"
     gene            5011..5213
                     /gene="TEF"
                     /label=TEF
     terminator      5016..5213
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     regulatory      complement(5272..5291)
                     /regulatory_class="promoter"
                     /note="T3 promoter"
     promoter        complement(5273..5291)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5308..5326)
                     /label=M13 reverse primer
                     /note="M13 reverse primer"
     primer_bind     complement(5312..5328)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5336..5352
                     /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)."
     promoter        complement(5360..5390)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(5532..6412)
                     /direction=LEFT
                     /label=ColE1 ori
                     /note="ColE1 ori"
     rep_origin      complement(5714..6302)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6473..7333)
                     /codon_start=1
                     /gene="ampR"
                     /product="beta-lactamase"
                     /label=ampR
                     /protein_id="AGL80166.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(6473..7333)
                     /gene="ampR"
                     /label=ampR
                     /note="bla"
     promoter        complement(7334..7438)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7475..7978
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     misc_feature    7475..7591
                     /label=yeast centromere CEN6
                     /note="yeast centromere CEN6"
     rep_origin      7592..7966
                     /label=yeast replication origin ARSH4
                     /note="yeast replication origin ARSH4"