LOCUS       Exported                3211 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pFF749, complete sequence.
ACCESSION   KM923745
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3211)
  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 3211)
  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 3211)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3211
                     /organism="Cloning vector pFF749"
                     /mol_type="other DNA"
                     /db_xref="taxon:1572888"
     regulatory      15..88
                     /regulatory_class="promoter"
     protein_bind    17..33
                     /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)."
     protein_bind    40..56
                     /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)."
     misc_feature    97..178
                     /note="primer for the Ec86 RT; msr"
     misc_feature    complement(168..298)
                     /note="template for Ec86 RT; msd(kanR_ON)"
     CDS             318..1280
                     /codon_start=1
                     /product="retron EC86 RNA-directed DNA polymerase-like 
                     protein"
                     /label=retron EC86 RNA-directed DNA polymerase-like pr
                     /protein_id="AIZ09105.1"
                     /translation="MKSAEYLNTFRLRNLGLPVMNNLHDMSKATRISVETLRLLIYTAD
                     FRYRIYTVEKKGPEKRMRTIYQPSRELKALQGWVLRNILDKLSSSPFSIGFEKHQSILN
                     NATPHIGANFILNIDLEDFFPSLTANKVFGVFHSLGYNRLISSVLTKICCYKNLLPQGA
                     PSSPKLANLICSKLDYRIQGYAGSRGLIYTRYAAALTLSAQSMKKVVKARDFLFSIIPS
                     EGLVINSKKTCISGPRSQRKVTGLVISQEKVGIGREKYKEIRAKIHHIFCGKSSEIEHV
                     RGWLSFILSVDSKSHRRLITYISKLEKKYGKNPLNKAKT"
     regulatory      1305..1409
                     /regulatory_class="terminator"
     terminator      1310..1396
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      1552..2222
                     /label=colE1
                     /note="colE1"
     rep_origin      complement(1560..2148)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      2236..2330
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2351..3010)
                     /codon_start=1
                     /product="CmR"
                     /label=CmR
                     /protein_id="AIZ09106.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(3011..3113)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"