LOCUS       Exported                5821 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pFNK-804-lacO-lacI, complete sequence.
ACCESSION   MH300676
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5821)
  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 5821)
  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 5821)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5821
                     /organism="Cloning vector pFNK-804-lacO-lacI"
                     /mol_type="other DNA"
                     /db_xref="taxon:2231588"
     CDS             complement(44..769)
                     /codon_start=1
                     /product="RhlR IS3-3"
                     /label=RhlR IS3-3
                     /protein_id="AWV91938.1"
                     /translation="MRNDGGFLLWWDGLRSEMQPIHDSQGVFAVLEKEVRRLGFDYYAY
                     GVRHTIPFTRPKTEVHGTYPKAWLERYQMQNYGAVDPAILNGLRSSEMVVWSDSLFDQS
                     RMLWNEARDWGLCVGATLPFRAPNNLLSVLSVARDQQNISSFEREEIRLRLRCMIELLT
                     QKLTDLEHPMLMSNPVCLSHREREILQWTADGKSSGEIAIILSISESTVNFHHKNIQKK
                     FDAPNKTLAAAYAAALGLI"
     RBS             776..787
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     regulatory      complement(779..787)
                     /regulatory_class="ribosome_binding_site"
                     /note="RBSII"
     regulatory      complement(802..844)
                     /regulatory_class="promoter"
     regulatory      complement(812..817)
                     /regulatory_class="minus_10_signal"
     regulatory      complement(834..840)
                     /regulatory_class="minus_35_signal"
     regulatory      857..939
                     /regulatory_class="promoter"
                     /note="prokaryotic; qsc119-lacO"
     regulatory      919..939
                     /regulatory_class="transcriptional_cis_regulatory_region"
                     /note="lacO"
     protein_bind    921..937
                     /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      946..962
                     /regulatory_class="ribosome_binding_site"
                     /note="RBSH"
     CDS             963..1715
                     /codon_start=1
                     /product="cI(LVA)"
                     /label=cI
                     /protein_id="AWV91939.1"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSIEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFGRPAANDENYALVA"
     CDS             963..1673
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=-lambda repressor
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSIEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     misc_feature    1674..1712
                     /label=Region: LVA
                     /note="Region: LVA"
     RBS             1720..1731
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     regulatory      1720..1728
                     /regulatory_class="ribosome_binding_site"
                     /note="RBSII"
     CDS             1738..2493
                     /codon_start=1
                     /product="GFP(LVA)"
                     /label=GFP
                     /protein_id="AWV91940.1"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYKRPAANDENYALVA"
     CDS             1738..2451
                     /codon_start=1
                     /product="Aequoria victoria green fluorescent protein"
                     /label=GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     regulatory      2709..2813
                     /regulatory_class="terminator"
                     /note="T1"
     terminator      2713..2799
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2886..3968)
                     /codon_start=1
                     /product="LacI"
                     /label=LacI
                     /protein_id="AWV91941.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(3969..4046)
                     /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(3995..4050)
                     /regulatory_class="promoter"
     regulatory      complement(4004..4009)
                     /regulatory_class="minus_10_signal"
     regulatory      complement(4027..4032)
                     /regulatory_class="minus_35_signal"
     rep_origin      4141..4703
                     /label=p15A ori
                     /note="p15A ori"
     rep_origin      complement(4143..4688)
                     /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."
     regulatory      complement(4797..4902)
                     /regulatory_class="terminator"
                     /note="T0"
     terminator      4802..4896
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4927..5721)
                     /codon_start=1
                     /product="Kan(r)"
                     /label=Kan
                     /protein_id="AWV91942.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"