pKD227 vector (V005250)

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LOCUS       40924_26596        5596 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pKD227, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5596)
  AUTHORS   Landry BP, Palanki R, Dyulgyarov N, Hartsough LA, Tabor JJ.
  TITLE     Phosphatase activity tunes two-component system sensor detection 
            threshold
  JOURNAL   Nat Commun 9 (1), 1433 (2018)
  PUBMED    29650958
REFERENCE   2  (bases 1 to 5596)
  AUTHORS   Landry BP.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-OCT-2017) Bioengineering, Rice University, 6100 Main 
            St, Houston, TX 77005, USA
REFERENCE   3  (bases 1 to 5596)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5596)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun"; date: "2018"; volume: "9"; issue: "1"; pages: "1433"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (29-OCT-2017) Bioengineering, Rice University, 6100 Main St, 
            Houston, TX 77005, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5596
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..148
                     /label=pKanR from E. coli tn5
                     /note="pKanR from E. coli tn5"
                     /regulatory_class="promoter"
     CDS             149..937
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     terminator      952..1046
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     misc_feature    1048..1049
                     /label=GC>CA
                     /note="GC>CA"
     rep_origin      1160..1705
                     /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."
     promoter        1878..1955
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             1956..3035
                     /label=lacI
                     /note="lac repressor"
     terminator      3044..3130
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     misc_feature    complement(3144..3149)
                     /label=AvrII Scar
                     /note="AvrII Scar"
     misc_feature    complement(3149..3196)
                     /label=ColE1 Fragment
                     /note="ColE1 Fragment"
     misc_feature    3258
                     /label=+1
                     /note="+1"
     promoter        3302..3330
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     misc_feature    3336
                     /label=+1
                     /note="+1"
     protein_bind    3338..3354
                     /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)."
     misc_feature    3364..3397
                     /label=sRBS (1096.98)
                     /note="sRBS (1096.98)"
     CDS             3398..5413
                     /codon_start=1
                     /product="Sbal195_3859 (TtrS)"
                     /label=Sbal195_3859
                     /protein_id="AWU50966.1"
                     /translation="MFSSGMSLKWPLSIQGSIEPKMTPRIIFRMALRTKVFITLLACLF
                     ASANVFAVEPQNQSAPSDVMDAEVVAPVATEEPGYQVVDVGVLAIRGYKATINRWQPLM
                     GWLETQIPNSYFRLHPLSLDELAKGVETQGLDFVITNPGQSVLLARQYSLTWLATLRSP
                     LNNGAAMQVGSALVVRADSPYQTLTDLKGRRLGIVSKNAFGGYLTLVYEAQLKGIDLPR
                     FVGEIIPLGFPLDNLLYQLDDQKSGNDAAKDNRLDATVVPVCQLEQMQAEGLINIGHYR
                     VLDNQTPVGFHCQVSTRLYPNWSMAKTNRASQSLAKSVTQALLALPEDHLAAKSSDSAG
                     WTTAVSQLAIDQLLKDLDMHPLQTPWWQRAWQWVKLHQQWAWFILGILVLLNAYHFWLE
                     YRFSRRGRELINTQRQLNENRALLEHAQRIAIAGELGASLSHELNQPLAAIGHYCHGAE
                     VRLQRGTSPEELQSVLTLIQQEVTRADSIISRLRNLLKKRPVSKQALYLHELVNETVPL
                     LAYEFEQHQINLAVNVSGEPYLQSLDEVGMQQLLLNLLKNAFDACVQRLELESSGTEQS
                     GMRKPYVPTIDIDLRYQERTLLLTVTDNGTGLTEETSLLMQAFYSTKSEGLGLGLVICR
                     DIAESHGGTFSLESAMGGGCQAQVAIPRKPEPNGVL"
     misc_feature    3503..3557
                     /label=TM
                     /note="TM"
     regulatory      4490..4499
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    4532..4588
                     /label=TM
                     /note="TM"
     misc_feature    4718..4720
                     /label=Catalytic His
                     /note="Catalytic His"
     terminator      complement(5434..5477)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     misc_feature    5503..5510
                     /label=BB Scar
                     /note="BB Scar"
     terminator      5518..5545
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    5552..5596
                     /label=pUC19 MCS
                     /note="pUC19 MCS"