LOCUS       Exported                9609 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Load vector pLD02X, complete sequence.
ACCESSION   KM457487
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9609)
  AUTHORS   Mishra D, Rivera PM, Lin A, Del Vecchio D, Weiss R.
  TITLE     A load driver device for engineering modularity in biological 
            networks
  JOURNAL   Nat. Biotechnol. 32 (12), 1268-1275 (2014)
  PUBMED    25419739
REFERENCE   2  (bases 1 to 9609)
  AUTHORS   Mishra D, Rivera-Ortiz PM, Lin A, Del Vecchio D, Weiss R.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-SEP-2014) Synthetic Biology Center, MIT, 500 
            Technology Square, NE47, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 9609)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9609
                     /organism="Load vector pLD02X"
                     /mol_type="other DNA"
                     /db_xref="taxon:1565121"
     regulatory      34..223
                     /regulatory_class="terminator"
                     /note="CYC1 terminator"
     terminator      34..223
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     rep_origin      410..1077
     rep_origin      complement(466..1054)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1225..2085)
                     /codon_start=1
                     /gene="ampR"
                     /product="AmpR"
                     /label=ampR
                     /protein_id="AIU94968.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(1225..2085)
                     /gene="ampR"
                     /label=ampR
     promoter        complement(2086..2190)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2217..3559
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     rep_origin      2219..3374
     protein_bind    complement(3180..3227)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        3828..4043
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4044..4847
                     /codon_start=1
                     /gene="URA3"
                     /product="URA3"
                     /label=URA3
                     /protein_id="AIU94970.1"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     gene            4044..4847
                     /gene="URA3"
                     /label=URA3
     rep_origin      complement(4978..5433)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      complement(5110..5416)
                     /direction=LEFT
     terminator      5512..5677
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     regulatory      complement(5512..5676)
                     /regulatory_class="terminator"
     misc_feature    5917..5938
                     /label=attB4
                     /note="attB4"
     protein_bind    5918..5938
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     regulatory      5972..6704
                     /regulatory_class="promoter"
     misc_feature    6388..6427
                     /label=SKN7 binding site
                     /note="SKN7 binding site"
     misc_feature    6428..6467
                     /label=SKN7 binding site
                     /note="SKN7 binding site"
     protein_bind    6716..6740
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    6716..6737
                     /label=attP1
                     /note="attP1"
     primer_bind     6886..6902
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6907..6931)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    complement(6909..6930)
                     /label=attB2
                     /note="attB2"
     regulatory      6936..7147
                     /regulatory_class="terminator"
     terminator      6954..7141
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     misc_feature    7244..7265
                     /label=attB4
                     /note="attB4"
     protein_bind    7245..7265
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     regulatory      7299..8031
                     /regulatory_class="promoter"
     misc_feature    7715..7754
                     /label=SKN7 binding site
                     /note="SKN7 binding site"
     misc_feature    7755..7794
                     /label=SKN7 binding site
                     /note="SKN7 binding site"
     protein_bind    8043..8067
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    8043..8064
                     /label=attP1
                     /note="attP1"
     primer_bind     8213..8229
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(8234..8258)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    complement(8236..8257)
                     /label=attB2
                     /note="attB2"
     regulatory      8263..8524
                     /regulatory_class="terminator"
     terminator      8273..8520
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     CDS             8681..9490
                     /codon_start=1
                     /gene="kanR"
                     /product="KanR"
                     /label=kanR
                     /protein_id="AIU94969.1"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            8681..9490
                     /gene="kanR"
                     /label=kanR