LOCUS       Exported                4534 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pECT3, complete sequence.
ACCESSION   KM521207
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4534)
  AUTHORS   Cameron DE, Collins JJ.
  TITLE     Tunable protein degradation in bacteria
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4534)
  AUTHORS   Cameron DE, Collins JJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-SEP-2014) Biomedical Engineering, Boston University, 
            44 Cummington St., Boston, MA 02215, USA
REFERENCE   3  (bases 1 to 4534)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4534
                     /organism="Cloning vector pECT3"
                     /mol_type="other DNA"
                     /db_xref="taxon:1565071"
     misc_feature    142..225
                     /label=pdt#3
                     /note="pdt#3"
     primer_bind     142..159
                     /label=P1 primer
                     /note="P1 primer"
     misc_feature    276..309
                     /label=FRT site
                     /note="FRT site"
     protein_bind    276..309
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             677..1471
                     /codon_start=1
                     /product="KanR"
                     /label=KanR
                     /note="kanamycin resistance"
                     /protein_id="AIU94516.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     protein_bind    1484..1531
                     /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)."
     misc_feature    1498..1531
                     /label=FRT site
                     /note="FRT site"
     primer_bind     complement(1535..1553)
                     /label=P2 primer
                     /note="P2 primer"
     CDS             complement(2344..3204)
                     /codon_start=1
                     /product="bla"
                     /label=bla
                     /note="ampicillin resistance"
                     /protein_id="AIU94517.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3205..3309)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3411..3830
                     /label=R6K origin
                     /note="R6K origin"
     rep_origin      3419..3807
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     rep_origin      complement(3870..4325)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     4467..4483
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4493..4511
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"