LOCUS       Exported                5818 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pFNK-806, complete sequence.
ACCESSION   MH300677
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5818)
  AUTHORS   Karig D, Martini KM, Lu T, DeLateur NA, Goldenfeld N, Weiss R.
  TITLE     Stochastic Turing patterns in a synthetic bacterial population
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. (2018) In press
  PUBMED    29891706
REFERENCE   2  (bases 1 to 5818)
  AUTHORS   Karig DK, Lu T.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-MAY-2018) Research and Development Department, Johns 
            Hopkins University Applied Physics Laboratory, 11100 Johns Hopkins 
            Rd, Laurel, MD 20723, USA
REFERENCE   3  (bases 1 to 5818)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5818
                     /organism="Cloning vector pFNK-806"
                     /mol_type="other DNA"
                     /db_xref="taxon:2231589"
     regulatory      complement(35..140)
                     /regulatory_class="terminator"
                     /note="T0"
     terminator      40..134
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(165..959)
                     /codon_start=1
                     /product="Kan(r)"
                     /label=Kan
                     /protein_id="AWV91943.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(1103..1828)
                     /codon_start=1
                     /product="RhlR IS3-3"
                     /label=RhlR IS3-3
                     /protein_id="AWV91944.1"
                     /translation="MRNDGGFLLWWDGLRSEMQPIHDSQGVFAVLEKEVRRLGFDYYAY
                     GVRHTIPFTRPKTEVHGTYPKAWLERYQMQNYGAVDPAILNGLRSSEMVVWSDSLFDQS
                     RMLWNEARDWGLCVGATLPFRAPNNLLSVLSVARDQQNISSFEREEIRLRLRCMIELLT
                     QKLTDLEHPMLMSNPVCLSHREREILQWTADGKSSGEIAIILSISESTVNFHHKNIQKK
                     FDAPNKTLAAAYAAALGLI"
     regulatory      complement(1829..1847)
                     /regulatory_class="ribosome_binding_site"
                     /note="RBS G"
     regulatory      complement(1848..1897)
                     /regulatory_class="promoter"
     regulatory      complement(1861..1866)
                     /regulatory_class="minus_10_signal"
     regulatory      complement(1885..1890)
                     /regulatory_class="minus_35_signal"
     regulatory      1975..1995
                     /regulatory_class="transcriptional_cis_regulatory_region"
                     /note="lacO"
     protein_bind    1977..1993
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     regulatory      2002..2018
                     /regulatory_class="ribosome_binding_site"
                     /note="RBSH"
     CDS             2019..2771
                     /codon_start=1
                     /product="cI(LVA)"
                     /label=cI
                     /protein_id="AWV91945.1"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSIEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFGRPAANDENYALVA"
     CDS             2019..2729
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=-lambda repressor
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSIEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     misc_feature    2730..2768
                     /label=Region: LVA
                     /note="Region: LVA"
     RBS             2776..2787
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     regulatory      2776..2784
                     /regulatory_class="ribosome_binding_site"
                     /note="RBSII"
     CDS             2794..3549
                     /codon_start=1
                     /product="GFP(LVA)"
                     /label=GFP
                     /protein_id="AWV91946.1"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYKRPAANDENYALVA"
     CDS             2794..3507
                     /codon_start=1
                     /product="Aequoria victoria green fluorescent protein"
                     /label=GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     regulatory      3765..3869
                     /regulatory_class="terminator"
                     /note="T1"
     terminator      3769..3855
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3942..5024)
                     /codon_start=1
                     /product="LacI"
                     /label=LacI
                     /protein_id="AWV91947.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(5025..5102)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     regulatory      complement(5051..5106)
                     /regulatory_class="promoter"
     regulatory      complement(5060..5065)
                     /regulatory_class="minus_10_signal"
     regulatory      complement(5083..5088)
                     /regulatory_class="minus_35_signal"
     rep_origin      5197..5759
                     /label=p15A ori
                     /note="p15A ori"
     rep_origin      complement(5199..5744)
                     /direction=LEFT
                     /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."