LOCUS       Exported                5193 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pWAS-anti-yfiD-anti-purB, complete sequence.
ACCESSION   MH488949
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5193)
  AUTHORS   Kim B, Binkley R, Kim HU, Lee SY.
  TITLE     Metabolic engineering of Escherichia coli for the enhanced 
            production of l-tyrosine
  JOURNAL   Biotechnol. Bioeng. (2018) In press
  PUBMED    30019750
REFERENCE   2  (bases 1 to 5193)
  AUTHORS   Yang D, Kim WJ, Yoo SM, Choi JH, Ha SH, Lee MH, Lee SY.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-JUN-2018) Dept. Chemical and Biomolecular Engineering,
            Korea Advanced Institute of Science and Technology, Daehak-ro 291, 
            Daejeon 34141, South Korea
REFERENCE   3  (bases 1 to 5193)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5193
                     /organism="Cloning vector pWAS-anti-yfiD-anti-purB"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /db_xref="taxon:2301190"
     rep_origin      complement(281..895)
                     /direction=LEFT
                     /label=ColE1
                     /note="ColE1"
     rep_origin      292..880
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      complement(1117..1245)
                     /regulatory_class="terminator"
                     /note="T1/TE"
     terminator      1123..1150
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      1166..1237
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     ncRNA           1252..1330
                     /product="micC"
                     /label=antisense_RNA
                     /ncRNA_class="antisense_RNA"
     misc_binding    complement(1331..1354)
                     /label=anti-yfiD binding site
                     /bound_moiety="anti-yfiD"
     regulatory      complement(1355..1403)
                     /regulatory_class="promoter"
                     /note="lambda PR promoter"
     primer_bind     complement(1423..1439)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      1446..1473
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      1489..1560
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1575..2288)
                     /codon_start=1
                     /gene="cI(ts2)"
                     /product="lambda cI repressor (temperature sensitive)"
                     /label=cI(ts2)
                     /protein_id="AXN70072.1"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPKLRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGEVIASQWPEETFG"
     gene            complement(1575..2288)
                     /gene="cI(ts2)"
                     /label=cI(ts2)
     promoter        complement(2331..2615)
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     regulatory      complement(2331..2365)
                     /regulatory_class="promoter"
                     /note="pBAD"
     CDS             2642..3520
                     /codon_start=1
                     /gene="araC"
                     /product="arabinose operon regulatory protein"
                     /label=araC
                     /protein_id="AXN70073.1"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     gene            2642..3520
                     /gene="araC"
                     /label=araC
     regulatory      3712..3760
                     /regulatory_class="promoter"
                     /note="lambda PR promoter"
     misc_binding    3761..3784
                     /label=anti-purB binding site
                     /bound_moiety="anti-purB"
     ncRNA           3785..3863
                     /product="micC"
                     /label=antisense_RNA
                     /ncRNA_class="antisense_RNA"
     regulatory      3870..3998
                     /regulatory_class="terminator"
                     /note="T1/TE"
     terminator      3878..3949
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3965..3992
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        4050..4154
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4155..5015
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="AXN70074.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            4155..5015
                     /gene="bla"
                     /label=bla