pEMY128AD_pY128 vector (V007567)

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LOCUS       40924_17394        4928 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pEMY128AD_pY128, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4928)
  AUTHORS   Young EM, Zhao Z, Gielesen BE, Wu L, Benjamin Gordon D, Roubos JA, 
            Voigt CA.
  TITLE     Iterative algorithm-guided design of massive strain libraries, 
            applied to itaconic acid production in yeast
  JOURNAL   Metab. Eng. 48, 33-43 (2018)
  PUBMED    29753070
REFERENCE   2  (bases 1 to 4928)
  AUTHORS   Young E, Zhao Z, Gielesen B, Wu L, Gordon DB, Roubos J, Voigt C.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-MAY-2018) Genetics, DSM Biotechnology Center, 
            Alexander Fleminglaan 1, Delft, South Holland 2613AX, Netherlands
REFERENCE   3  (bases 1 to 4928)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4928)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Metab. Eng.
            48, 33-43 (2018)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (21-MAY-2018) Genetics, DSM Biotechnology Center, Alexander 
            Fleminglaan 1, Delft, South Holland 2613AX, Netherlands"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4928
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     229..245
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    246..302
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(315..331)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(339..355)
                     /label=lac operator
                     /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(363..393)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(408..429)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(659..1247)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      1346..2505
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     gene            complement(2547..3903)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin
                     resistance (Wach et al., 1994)"
     CDS             complement(4075..4731)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(4732..4835)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     terminator      complement(4894..4923)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"