LOCUS       Exported                4160 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pFF746, complete sequence.
ACCESSION   KM923748
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4160)
  AUTHORS   Farzadfard F, Lu TK.
  TITLE     Synthetic biology. Genomically encoded analog memory with precise in
            vivo DNA writing in living cell populations
  JOURNAL   Science 346 (6211), 1256272 (2014)
  PUBMED    25395541
REFERENCE   2  (bases 1 to 4160)
  AUTHORS   Farzadfard F, Lu T.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-DEC-2014) Biological Engineering, MIT, 77 
            Massachusetts Avenue, NE47, Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 4160)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4160
                     /organism="Cloning vector pFF746"
                     /mol_type="other DNA"
                     /db_xref="taxon:1572887"
     regulatory      7..80
                     /regulatory_class="promoter"
     promoter        7..80
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    7..25
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    32..50
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    89..170
                     /note="primer for the Ec86 RT; msr"
     misc_feature    complement(160..293)
                     /note="template for Ec86 RT; msd(galK_ON)"
     CDS             313..1275
                     /codon_start=1
                     /product="retron EC86 RNA-directed DNA polymerase-like 
                     protein"
                     /label=retron EC86 RNA-directed DNA polymerase-like pr
                     /protein_id="AIZ09112.1"
                     /translation="MKSAEYLNTFRLRNLGLPVMNNLHDMSKATRISVETLRLLIYTAD
                     FRYRIYTVEKKGPEKRMRTIYQPSRELKALQGWVLRNILDKLSSSPFSIGFEKHQSILN
                     NATPHIGANFILNIDLEDFFPSLTANKVFGVFHSLGYNRLISSVLTKICCYKNLLPQGA
                     PSSPKLANLICSKLDYRIQGYAGSRGLIYTRYADDLTLSAQSMKKVVKARDFLFSIIPS
                     EGLVINSKKTCISGPRSQRKVTGLVISQEKVGIGREKYKEIRAKIHHIFCGKSSEIEHV
                     RGWLSFILSVDSKSHRRLITYISKLEKKYGKNPLNKAKT"
     CDS             1312..2097
                     /codon_start=1
                     /product="bet protein"
                     /label=bet protein
                     /note="recombination"
                     /protein_id="AIZ09114.1"
                     /translation="MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFI
                     ALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESC
                     TCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLA
                     FGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLP
                     LCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA"
     regulatory      2110..2214
                     /regulatory_class="terminator"
     terminator      2115..2201
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      2222..3017
                     /label=p15A
                     /note="p15A"
     rep_origin      complement(2371..2916)
                     /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      3030..3124
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3155..3949)
                     /codon_start=1
                     /product="kanamycin phosphotransferase"
                     /label=kanamycin phosphotransferase
                     /protein_id="AIZ09113.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     regulatory      complement(4038..4087)
                     /regulatory_class="promoter"