LOCUS       Exported                9286 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pECGMC3, complete sequence.
ACCESSION   KM521211
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9286)
  AUTHORS   Cameron DE, Collins JJ.
  TITLE     Tunable protein degradation in bacteria
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9286)
  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 9286)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..9286
                     /organism="Cloning vector pECGMC3"
                     /mol_type="other DNA"
                     /db_xref="taxon:1565075"
     regulatory      1..276
                     /regulatory_class="promoter"
                     /note="P32 promoter"
     regulatory      226..231
                     /regulatory_class="minus_35_signal"
                     /note="-35 binding site"
     regulatory      249..254
                     /regulatory_class="minus_10_signal"
                     /note="-10 binding site"
     CDS             304..1011
                     /codon_start=1
                     /product="mcherry"
                     /label=mcherry
                     /note="L. lactis optimized"
                     /protein_id="AIU94528.1"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     misc_feature    1012..1095
                     /label=pdt#3
                     /note="pdt#3"
     regulatory      1114..1218
                     /regulatory_class="terminator"
                     /note="T1 terminator"
     terminator      1119..1205
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      1225..2033
                     /label=colE1
                     /note="colE1"
     rep_origin      complement(1370..1958)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      2040..2145
                     /regulatory_class="terminator"
                     /note="terminator T0"
     terminator      2046..2140
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2170..3030)
                     /codon_start=1
                     /product="bla"
                     /label=bla
                     /note="ampicillin resistance"
                     /protein_id="AIU94529.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3031..3135)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3167..5714)
                     /direction=LEFT
                     /label=AMB1
                     /note="AMB1"
     CDS             complement(5732..6469)
                     /codon_start=1
                     /product="emR"
                     /label=emR
                     /note="erythromycin resistance"
                     /protein_id="AIU94530.1"
                     /translation="MNKNIKYSQNFLTSEKVLNQIIKQLNLKETDTVYEIGTGKGHLTT
                     KLAKISKQVTSIELDSHLFNLSSEKLKLNTRVTLIHQDILQFQFPNKQRYKIVGSIPYH
                     LSTQIIKKVVFESHASDIYLIVEEGFYKRTLDIHRTLGLLLHTQVSIQQLLKLPAECFH
                     PKPKVNSVLIKLTRHTTDVPDKYWKLYTYFVSKWVNREYRQLFTKNQFHQAMKHAKVNN
                     LSTVTYEQVLSIFNSYLLFNGRK"
     regulatory      complement(6647..6691)
                     /regulatory_class="terminator"
                     /note="terminator"
     terminator      6655..6682
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(6723..9086)
                     /codon_start=1
                     /product="mf-lon"
                     /label=mf-lon
                     /protein_id="AIU94531.1"
                     /translation="MSKKIKLPIFQIRGSFIVPGIKENLEVGRKNTLASVNYAIKNSNN
                     QMIAIPQIDASVEKPEFSDLHEFGILIDFEVIKEWKDNSLTISTNPIQRCKVISFFENE
                     DQVPYAEVELIESINDFSDEELKELIEKISDAIKTKASLVTKQIKQLISGESDDLSLAF
                     DSIMFKLAPSKILTNPEYITSPSLKTRWSIIEKIIFAEDGIITRNAESIDAARQKNEIE
                     QELNHKLKEKMDKQQKEYYLREKMRIIKDELEDEDDSDDSSLEKYKERLAKEPFPEEVK
                     RKIMASIKRVEALQSGTPEWNTEKNYIDWMMSIPWWEETEDLTDLKYAKKILDKHHYGM
                     KKVKERIIEYLAVKTKTKSLKAPIITLVGPPGVGKTSLAKSIAEAVGKNFVKVSLGGVK
                     DESEIRGHRKTYVGSMPGRIIQTMKRAKVKNPLFLLDEIDKMASDHRGDPASAMLEVLD
                     PEQNKEFSDHYIEEPYDLSQVMFIATANYPEDIPEALYDRMEIINLSSYTEIEKVKIAQ
                     DYLVPKAIEQHELTSEEISFTEGAINEIIKYYTREAGVRQLERHINSIIRKYIVKNLNG
                     EMDKIVIDEKQVNDLLGKRIFDHTEKQEESQIGVVTGLAYTQFGGDILPIEVSLYPGKG
                     NLILTGKLGEVMKESATIALTYVKSNFEKFGVDKKVFEENDIHVHVPEGAVPKDGPSAG
                     ITITTALISALSDKPVSKEIGMTGEITLRGNVLPIGGLREKSISASRSGLKTIIIPKKN
                     ERDLDEIPDEVKAKLKIIPAEKYEEVFAIVFKTK"
     regulatory      complement(9105..9280)
                     /regulatory_class="promoter"
                     /note="nisA promoter"
     regulatory      complement(9132..9137)
                     /regulatory_class="minus_10_signal"
                     /note="-10 region"
     regulatory      complement(9158..9163)
                     /regulatory_class="minus_35_signal"
                     /note="-35 region"