LOCUS       Exported                6058 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Shuttle expression vector pCK320, complete sequence.
ACCESSION   MG030637
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6058)
  AUTHORS   Kelly CL, Taylor GM, Hitchcock A, Torres-Mendez A, Heap JT.
  TITLE     A Rhamnose-Inducible System for Precise and Temporal Control of Gene
            Expression in Cyanobacteria
  JOURNAL   ACS Synth Biol (2018) In press
  PUBMED    29544054
REFERENCE   2  (bases 1 to 6058)
  AUTHORS   Kelly CL, Taylor GM, Mendez AT, Hitchcock A, Heap JT.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-OCT-2017) Centre for Synthetic Biology and Innovation,
            Department of Life Sciences, Imperial College London, South 
            Kensington Campus, London SW7 2AZ, United Kingdom
REFERENCE   3  (bases 1 to 6058)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6058
                     /organism="Shuttle expression vector pCK320"
                     /mol_type="other DNA"
                     /db_xref="taxon:2162677"
     misc_recomb     1..1015
                     /label=similar to ndhB
                     /note="similar to ndhB"
     regulatory      1022..1143
                     /regulatory_class="promoter"
                     /note="rhamnose"
     misc_binding    1038..1053
                     /label=CRP binding site
                     /bound_moiety="CRP"
     misc_binding    1057..1073
                     /label=RhaS binding site
                     /bound_moiety="RhaS"
                     /note="operator 1"
     misc_binding    1090..1105
                     /label=RhaS binding site
                     /bound_moiety="RhaS"
                     /note="operator 2"
     regulatory      1103..1108
                     /regulatory_class="minus_35_signal"
                     /note="PrhaBAD"
     regulatory      1127..1132
                     /regulatory_class="minus_10_signal"
                     /note="PrhaBAD"
     regulatory      1144..1177
                     /regulatory_class="ribosome_binding_site"
                     /note="synthetic"
     CDS             1178..1897
                     /codon_start=1
                     /product="YFP"
                     /label=YFP
                     /note="BBa_E0030"
                     /protein_id="AVX33632.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      1904..2013
                     /regulatory_class="promoter"
                     /note="kan"
     CDS             2014..2829
                     /codon_start=1
                     /product="KanR"
                     /label=KanR
                     /protein_id="AVX33633.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     regulatory      2830..2862
                     /regulatory_class="ribosome_binding_site"
                     /note="synthetic"
     CDS             2863..3699
                     /codon_start=1
                     /gene="rhaS"
                     /gene_synonym="ECK3898; JW3876; rhaC2"
                     /product="RhaS"
                     /label=rhaS
                     /note="positive regulator for rhaBAD operon; 
                     transcriptional activator of rhaBAD and rhaT"
                     /db_xref="ASAP:ABE-0012745"
                     /db_xref="EcoGene:EG10843"
                     /db_xref="UniProtKB/Swiss-Prot:P09377"
                     /db_xref="GeneID:948397"
                     /protein_id="AVX33634.1"
                     /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
                     GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
                     ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
                     ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
                     LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
     gene            2863..3699
                     /gene="rhaS"
                     /gene_synonym="ECK3898; JW3876; rhaC2"
                     /label=rhaS
                     /note="b3905"
                     /db_xref="EcoGene:EG10843"
                     /db_xref="GeneID:948397"
     misc_recomb     3700..4738
                     /label=similar to ndhB
                     /note="similar to ndhB"
     rep_origin      5054..5883
                     /label=p15A
                     /note="p15A"
     rep_origin      5071..5616
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."