LOCUS       Exported                4099 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pFNK-101, complete sequence.
ACCESSION   AY952935
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4099)
  AUTHORS   Basu S, Gerchman Y, Collins CH, Arnold FH, Weiss R.
  TITLE     A synthetic multicellular system for programmed pattern formation
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4099)
  AUTHORS   Basu S, Gerchman Y, Weiss R.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-FEB-2005) Electrical Engineering, Princeton 
            University, B-312, E-Quad, Olden Street, Princeton, NJ 08544, USA
REFERENCE   3  (bases 1 to 4099)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4099
                     /organism="Expression vector pFNK-101"
                     /mol_type="other DNA"
                     /db_xref="taxon:316974"
     CDS             complement(76..1158)
                     /codon_start=1
                     /gene="lacI"
                     /product="LacI"
                     /label=lacI
                     /note="Lac repressor"
                     /protein_id="AAX29982.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            complement(76..1158)
                     /gene="lacI"
                     /label=lacI
     promoter        complement(1159..1236)
                     /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(1185..1240)
                     /regulatory_class="promoter"
                     /note="PlacIq"
     regulatory      1290..1373
                     /regulatory_class="promoter"
                     /note="Plac; LacI repressed promotor"
     protein_bind    1302..1323
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1338..1368
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1376..1392
                     /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)."
     RBS             1403..1414
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1421..2107
                     /codon_start=1
                     /product="HcRed"
                     /label=HcRed
                     /note="red fluorescence protein from Heteractis crispa"
                     /protein_id="AAX29983.1"
                     /translation="MVSGLLKESMRIKMYMEGTVNGHYFKCEGEGDGNPFAGTQSMRIH
                     VTEGAPLPFAFDILAPCCEYGSRTFVHHTAEIPDFFKQSFPEGFTWERTTTYEDGGILT
                     AHQDTSLEGNCLIYKVKVHGTNFPADGPVMKNKSGGWEPSTEVVYPENGVLCGRNVMAL
                     KVGDRHLICHHYTSYRSKKAVRALTMPGFHFTDIRLQMLRKKKDEYFELYEASVARYSD
                     LPEKAN"
     terminator      2186..2272
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      2440..3002
                     /label=p15A
                     /note="p15A"
     rep_origin      complement(2442..2987)
                     /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."
     terminator      3101..3195
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3226..4020)
                     /codon_start=1
                     /gene="kanR"
                     /product="KanR"
                     /label=kanR
                     /note="kanamycin resistance protein"
                     /protein_id="AAX29984.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(3226..4020)
                     /gene="kanR"
                     /label=kanR