LOCUS       Exported                8509 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pFF706, complete sequence.
ACCESSION   KM923751
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8509)
  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 8509)
  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 8509)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8509
                     /organism="Cloning vector pFF706"
                     /mol_type="other DNA"
                     /db_xref="taxon:1572884"
     misc_feature    42..184
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(286..477)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1278..1895)
                     /codon_start=1
                     /product="FixJ"
                     /label=FixJ
                     /protein_id="AIZ09126.1"
                     /translation="MTTKGHIYVIDDDAAMRDSLNFLLDSAGFGVTLFDDAQAFLDALP
                     GLSFGCVVSDVRMPGLDGIELLKRMKAQQSPFPILIMTGHGDVPLAVEAMKLGAVDFLE
                     KPFEDDRLTAMIESAIRQAEPAAKSEAVAQDIAARVASLSPRERQVMEGLIAGLSNKLI
                     AREYDISPRTIEVYRANVMTKMQANSLSELVRLAMRAGMLND"
     CDS             complement(1892..3034)
                     /codon_start=1
                     /product="YF1"
                     /label=YF1
                     /protein_id="AIZ09121.1"
                     /translation="MASFQSFGIPGQLEVIKKALDHVRVGVVITDPALEDNPIVYVNQG
                     FVQMTGYETEEILGKNCRFLQGKHTDPAEVDNIRTALQNKEPVTVQIQNYKKDGTMFWN
                     ELNIDPMEIEDKTYFVGIQNDITEHQQTQARLQELQSELVHVSRLSAMGEMASALAHEL
                     NQPLAAISNYMKGSRRLLAGSSDPNTPKVESALDRAAEQALRAGQIIRRLRDFVARGES
                     EKRVESLSKLIEEAGALGLAGAREQNVQLRFSLDPGADLVLADRVQIQQVLVNLFRNAL
                     EAMAQSQRRELVVTNTPAADDMIEVEVSDTGSGFQDDVIPNLFQTFFTTKDTGMGVGLS
                     ISRSIIEAHGGRMWAESNASGGATFRFTLPAADEMIGGLA"
     promoter        complement(3035..3112)
                     /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      3407..3653
                     /regulatory_class="promoter"
                     /note="FixK2 promoter"
     RBS             3660..3671
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3678..4424
                     /codon_start=1
                     /product="cI-LVA"
                     /label=cI-LVA
                     /note="lambda cI-LVA"
                     /protein_id="AIZ09124.1"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFGAANDENYALVA"
     CDS             3678..4388
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=-lambda repressor
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     CDS             4389..4421
                     /codon_start=1
                     /product="C-terminal peptide that mediates degradation in 
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
                     /label=ssrA tag (LVA)
                     /note="mutant LVA variant that confers accelerated 
                     degradation under some conditions (Andersen et al., 1998)"
                     /translation="AANDENYALVA"
     terminator      4469..4540
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4556..4583
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     regulatory      4598..4646
                     /regulatory_class="promoter"
                     /note="lambda pR promoter"
     misc_feature    4655..4736
                     /note="primer for the Ec86 RT; msr"
     misc_feature    complement(4726..4856)
                     /note="template for Ec86 RT; msd(kanR_ON)"
     CDS             4876..5838
                     /codon_start=1
                     /product="retron EC86 RNA-directed DNA polymerase-like 
                     protein"
                     /label=retron EC86 RNA-directed DNA polymerase-like pr
                     /protein_id="AIZ09122.1"
                     /translation="MKSAEYLNTFRLRNLGLPVMNNLHDMSKATRISVETLRLLIYTAD
                     FRYRIYTVEKKGPEKRMRTIYQPSRELKALQGWVLRNILDKLSSSPFSIGFEKHQSILN
                     NATPHIGANFILNIDLEDFFPSLTANKVFGVFHSLGYNRLISSVLTKICCYKNLLPQGA
                     PSSPKLANLICSKLDYRIQGYAGSRGLIYTRYADDLTLSAQSMKKVVKARDFLFSIIPS
                     EGLVINSKKTCISGPRSQRKVTGLVISQEKVGIGREKYKEIRAKIHHIFCGKSSEIEHV
                     RGWLSFILSVDSKSHRRLITYISKLEKKYGKNPLNKAKT"
     CDS             5875..6660
                     /codon_start=1
                     /product="bet protein"
                     /label=bet protein
                     /note="recombination"
                     /protein_id="AIZ09123.1"
                     /translation="MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFI
                     ALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESC
                     TCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLA
                     FGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLP
                     LCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA"
     regulatory      6673..6777
                     /regulatory_class="terminator"
     terminator      6678..6764
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      6920..7590
                     /label=colE1
                     /note="colE1"
     rep_origin      complement(6928..7516)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      7604..7698
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(7719..8378)
                     /codon_start=1
                     /product="CmR"
                     /label=CmR
                     /protein_id="AIZ09125.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(8379..8481)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"