LOCUS       Exported                4848 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pECM1, complete sequence.
ACCESSION   KM521208
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4848)
  AUTHORS   Cameron DE, Collins JJ.
  TITLE     Tunable protein degradation in bacteria
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4848)
  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 4848)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4848
                     /organism="Cloning vector pECM1"
                     /mol_type="other DNA"
                     /db_xref="taxon:1565072"
     primer_bind     142..165
                     /label=P1M primer
                     /note="P1M primer"
     misc_feature    142..147
                     /label=MmeI
                     /note="MmeI"
     misc_feature    148..231
                     /label=pdt#1
                     /note="pdt#1"
     misc_feature    282..315
                     /label=FRT site
                     /note="FRT site"
     protein_bind    282..315
                     /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             683..1477
                     /codon_start=1
                     /product="KanR"
                     /label=KanR
                     /note="kanamycin resistance"
                     /protein_id="AIU94518.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     protein_bind    1490..1537
                     /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    1504..1537
                     /label=FRT site
                     /note="FRT site"
     primer_bind     complement(1545..1562)
                     /label=P2M primer
                     /note="P2M primer"
     misc_feature    1557..1562
                     /label=MmeI
                     /note="MmeI"
     CDS             complement(2658..3518)
                     /codon_start=1
                     /product="bla"
                     /label=bla
                     /note="ampicillin resistance"
                     /protein_id="AIU94519.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3519..3623)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3725..4144
                     /label=R6K origin
                     /note="R6K origin"
     rep_origin      3733..4121
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     rep_origin      complement(4184..4639)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     4781..4797
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4807..4825
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"