LOCUS       Exported                6996 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pDEST413TEFt7, complete sequence.
ACCESSION   JX901379
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6996)
  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 6996)
  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 6996)
  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 6996)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     On Jul 28, 2015 this sequence version replaced JX901379.1.
FEATURES             Location/Qualifiers
     source          1..6996
                     /organism="Cloning vector pDEST413TEFt7"
                     /mol_type="other DNA"
                     /db_xref="taxon:1333770"
     promoter        317..503
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             504..1163
                     /codon_start=1
                     /gene="his3"
                     /product="HIS3"
                     /label=his3
                     /note="derived from S. cerevisiae HIS3"
                     /protein_id="AGL80155.1"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     gene            504..1163
                     /gene="his3"
                     /label=his3
     rep_origin      complement(1423..1878)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      complement(1426..1881)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 ori"
     primer_bind     2023..2039
                     /label=M13 -20 primer
                     /note="M13 -20 primer"
     regulatory      2049..2068
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     promoter        2049..2067
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      complement(2096..2224)
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     terminator      2121..2168
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2166..2185
                     /label=T7 reverse primer
                     /note="T7 reverse primer"
     misc_recomb     2235..2359
                     /label=attR2 recombination site
                     /note="attR2 recombination site"
     protein_bind    2235..2359
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(2400..2705)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB"
                     /label=ccdB
                     /protein_id="AGL80156.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            complement(2400..2705)
                     /gene="ccdB"
                     /label=ccdB
     CDS             complement(3047..3706)
                     /codon_start=1
                     /gene="CmR"
                     /product="chloramphenicol resistance protein"
                     /label=CmR
                     /protein_id="AGL80157.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(3047..3706)
                     /gene="CmR"
                     /label=CmR
     promoter        complement(3760..3790)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     misc_recomb     complement(3815..3939)
                     /label=attR1 recombination site
                     /note="attR1 recombination site"
     protein_bind    complement(3815..3939)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      3960..4162
                     /regulatory_class="terminator"
                     /gene="TEF"
                     /note="derived from A. gossypii TEF terminator"
     gene            3960..4162
                     /gene="TEF"
                     /label=TEF
     terminator      3965..4162
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     regulatory      complement(4221..4240)
                     /regulatory_class="promoter"
                     /note="T3 promoter"
     promoter        complement(4222..4240)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4257..4275)
                     /label=M13 reverse primer
                     /note="M13 reverse primer"
     primer_bind     complement(4261..4277)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4285..4301
                     /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(4309..4339)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(4481..5361)
                     /direction=LEFT
                     /label=ColE1 ori
                     /note="ColE1 ori"
     rep_origin      complement(4663..5251)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5422..6282)
                     /codon_start=1
                     /gene="ampR"
                     /product="beta-lactamase"
                     /label=ampR
                     /protein_id="AGL80158.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(5422..6282)
                     /gene="ampR"
                     /label=ampR
                     /note="bla"
     promoter        complement(6283..6387)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    6424..6927
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     misc_feature    6424..6540
                     /label=yeast centromere CEN6
                     /note="yeast centromere CEN6"
     rep_origin      6541..6915
                     /label=yeast replication origin ARSH4
                     /note="yeast replication origin ARSH4"