pCF vector (V016410)

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LOCUS       62056_6220        8800 bp DNA     circular SYN 18-AUG-2021
DEFINITION  Expression vector pCF, complete sequence.
ACCESSION   MZ567118
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8800)
  AUTHORS   Fang L, Fan J, Luo S, Chen Y, Wang C, Cao Y, Song H.
  TITLE     Genome-scale target identification in Escherichia coli for 
            high-titer production of free fatty acids
  JOURNAL   Nat Commun 12 (1), 4976 (2021)
REFERENCE   2  (bases 1 to 8800)
  AUTHORS   Fang L, Fan J, Luo S, Chen Y, Wang C, Cao Y, Song H.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-JUL-2021) School of Chemical Engineering, Tianjin 
            University, No. 135, Yaguan Road, Haihe Education Park, Jinnan 
            District, Tianjin, Tian Jin, Tian Jin 300350, China
REFERENCE   3  (bases 1 to 8800)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun"; date: "2021"; volume: "12"; issue: "1"; pages: "4976"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (13-JUL-2021) School of Chemical Engineering, Tianjin University, 
            No. 135, Yaguan Road, Haihe Education Park, Jinnan District, 
            Tianjin, Tian Jin, Tian Jin 300350, China"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8800
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(120..776)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(777..879)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     rep_origin      complement(1405..1949)
                     /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."
     protein_bind    complement(2140..2161)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(2177..3256)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(3257..3334)
                     /label=lacI promoter
     promoter        3405..3434
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    3442..3458
                     /label=lac operator
                     /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    3475..3544
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region
                     (Berg et al., 1989)"
     CDS             3595..3618
                     /label=minicistron
                     /note="synthetic cistron containing a ribosome binding site
                     (Shine-Dalgarno sequence), for enhancing the bacterial 
                     expression of a downstream cistron (Schoner, 1997)"
     CDS             3625..7728
                     /label=dCas9
                     /note="catalytically dead mutant of the Cas9 endonuclease
                     from the Streptococcus pyogenes Type II CRISPR/Cas system"
     terminator      7780..7826
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     promoter        7898..7916
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    7917..7941
                     /label=lac operator
                     /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)."
     gene            7984..8535
                     /gene="tesA'"
                     /label=tesA'
     CDS             7984..8535
                     /codon_start=1
                     /transl_table=11
                     /gene="tesA'"
                     /product="TesA'"
                     /label=tesA'
                     /note="truncated fatty acyl-ACP thioesterase TesA"
                     /protein_id="QXV67745.1"
                     /translation="MADTLLILGDSLSAGYRMSASAAWPALLNDKWQSKTSVVNASISG
                     DTSQQGLARLPALLKQHQPRWVLVELGGNDGLRGFQPQQTEQTLRQILQDVKAANAEPL
                     LMQIRLPANYGRRYNEAFSAIYPKLAKEFDVPLLPFFMEEVYLKPQWMQDDGIHPNRDA
                     QPFIADWMAKQLQPLVNHDS"
     CDS             8554..8598
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     terminator      8650..8697
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"