pUDE214 vector (V002340)

LOCUS       40924_45458       11757 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pUDE214, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11757)
  AUTHORS   Kozak BU, van Rossum HM, Benjamin KR, Wu L, Daran JM, Pronk JT, van 
            Maris AJ.
  TITLE     Replacement of the Saccharomyces cerevisiae acetyl-CoA synthetases 
            by alternative pathways for cytosolic acetyl-CoA synthesis
  JOURNAL   Metab. Eng. (2013) In press
  PUBMED    24269999
REFERENCE   2  (bases 1 to 11757)
  AUTHORS   Kozak BU, van Rossum HM, Benjamin KR, Wu L, Daran J-M.G., Pronk JT, 
            van Maris AJA.
  TITLE     Direct Submission
  JOURNAL   Submitted (31-MAY-2013) Biotechnology, Delft University of 
            Technology, Julianalaan 67, Delft 2628BC, The Netherlands
REFERENCE   3  (bases 1 to 11757)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 11757)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Metab. Eng.
            (2013) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (31-MAY-2013) Biotechnology, Delft University of Technology, 
            Julianalaan 67, Delft 2628BC, The Netherlands"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: IDBA v. 0.20
            Sequencing Technology :: Illumina
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..11757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(183..771)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(945..1802)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(1803..1907)
                     /label=AmpR promoter
     rep_origin      complement(1934..3276)
                     /direction=LEFT
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        3538..3753
                     /label=URA3 promoter
     CDS             3754..4554
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     rep_origin      complement(4688..5143)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     5284..5300
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5310..5328
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        5838..6357
                     /label=TPI1 promoter
                     /note="strong constitutive promoter for yeast triose
                     phosphate isomerase"
     CDS             6358..8616
                     /codon_start=1
                     /gene="pflB"
                     /product="formate C-acetyltransferase"
                     /label=pflB
                     /protein_id="AHB33316.1"
                     /translation="MITSEKTTKPAAWKGFKGGHWQEEINIRDFIQNNFTQYNGDESFL
                     AGPTAATKTLNDKVLELKKQERAAGGVLDADTKVVATITSHGPGYIQKDLEKIVGLQTD
                     KPLKRAFMPFGGIRMADDALKSYGYTPDEENDKIFTEYRKTHNQGVFDVYTPDMRKARH
                     YKIITGLPDAYARGRLIPDLPRVAVYGIDRLMEDKANDFAHIGDGELTDDVIRLREEVQ
                     DQYRALADMKKMAASYGYDISKPATNAQEAIQWMYFAYLAAIKTQNGAAMSVGRIDTTM
                     DIFIQRDLDNGVLDESQAQELIDQFVMKLRMVRFIRTEEYNSLFSGDPIWATLSMCGLG
                     VDGQHHVTKTAFRILKTLDNMGAAPEPNITILWSDRLPEDFKRYATEVSIDSSTIQYEN
                     DDLMRVQWGTDYYGIACCVSAQPIADGIQYFGARANLAKAILYAINGGRDEIAGDQVGP
                     AYEPITSEYIDYDEFMKKLDKQMDWLADTYVNSLNAIHYMHDKYYYEAAQLALKNTDLD
                     RTFATGISGLSHAADSISAIKYGHVKVIRDERGIAVDFKADNDYPRYGNNDDRADDIAK
                     WLVKELYSKMNTHHLYRNAKLSTSVLTITSNVVYGKNTGTTPNGRQKGEPFSPGANPAY
                     GAEKSGALASLLSTAKLPYRYATDGISNTFGVTPNTLGHDLESRKDTLVNMLDGYMKND
                     GMHLNINVFNKDTLIDAQKHPEEYPTLTVRVSGYCVYFADLTKEQQDDVISRTFFESM"
     gene            6358..8616
                     /gene="pflB"
                     /label=pflB
     regulatory      8617..9117
                     /label=GND2 terminator
                     /note="GND2 terminator"
                     /regulatory_class="terminator"
     regulatory      9122..10122
                     /label=FBA1 promoter
                     /note="FBA1 promoter"
                     /regulatory_class="promoter"
     CDS             10123..10944
                     /codon_start=1
                     /gene="pflA"
                     /product="formate C-acetyltransferase activating enzyme"
                     /label=pflA
                     /protein_id="AHB33317.1"
                     /translation="MENKQVSTTQAAAKEPLIGYVHSIETFGSVDGPGIRYVAFLQGCH
                     MRCQYCHNPDTWKLNVGDQMTADEILEDAAKYRAFWGKTGGITVSGGESLVQIDFILDL
                     FEKAKAMNISTCLDTSGQPFTREQPFFDKFERLMKVTDISLVDIKHIDSAKHKQLTQYG
                     NENILDMIQYMAQHHDDMWIRHVLVPQRTDYDEDLKKLGDYIAKIPNDVVQKVEVLPYH
                     TLGVKKYHEMKIKYRLEGIESPTQDRVANAEKLLHTADYNGYKTWMPLPKL"
     gene            10123..10944
                     /gene="pflA"
                     /label=pflA
     regulatory      10945..11445
                     /label=PMA1 terminator
                     /note="PMA1 terminator"
                     /regulatory_class="terminator"
     primer_bind     complement(11446..11462)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(11499..11517)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(11538..11554)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    11562..11578
                     /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(11586..11616)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(11631..11652)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."