pCLF2 vector (V008451)

Basic Vector Information

Vector Name:
pCLF2
Antibiotic Resistance:
Ampicillin
Length:
3043 bp
Type:
PCR amplification vector
Replication origin:
R6K γ ori
Source/Author:
Santiviago CA, Blondel CJ.

pCLF2 vector Map

pCLF23043 bp6001200180024003000priming site P1FRTcat promotercatFRT (minimal)T7 promoterpriming site P2R6K gamma oriAmpRAmpR promoterrrnB T1 terminatorrrnB T2 terminator

pCLF2 vector Sequence

LOCUS       40924_11041        3043 bp DNA     circular SYN 17-DEC-2018
DEFINITION  PCR amplification vector pCLF2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3043)
  AUTHORS   Santiviago CA, Blondel CJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (31-MAR-2010) Laboratorio de Microbiologia, Departamento 
            de Bioquimica y Biologia Molecular, Facultad de Ciencias Quimicas y 
            Farmaceuticas, Universidad de Chile, Avenida Vicuna Mackenna 20, 
            Santiago, Region Metropolitana, Chile
REFERENCE   2  (bases 1 to 3043)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 3043)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (31-MAR-2010) Laboratorio de Microbiologia, Departamento de 
            Bioquimica y Biologia Molecular, Facultad de Ciencias Quimicas y 
            Farmaceuticas, Universidad de Chile, Avenida Vicuna Mackenna 20, 
            Santiago, Region Metropolitana, Chile"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3043
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     31..50
                     /label=priming site P1
                     /note="priming site P1"
     primer_bind     32..51
                     /label=priming site P3
                     /note="priming site P3"
     protein_bind    complement(51..98)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_feature    51..96
                     /note="FLP recognition target site; FRT"
     promoter        273..375
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             376..1014
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyl transferase"
                     /label=cat
                     /note="confers chloramphenicol resistance"
                     /protein_id="ADZ54044.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRCLMNTTVLR"
     gene            376..1014
                     /gene="cat"
                     /label=cat
     primer_bind     complement(470..492)
                     /label=priming site c3
                     /note="priming site c3"
     primer_bind     903..922
                     /label=priming site c1
                     /note="priming site c1"
     protein_bind    complement(1150..1183)
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     promoter        1203..1221
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1227..1246)
                     /label=priming site P2
                     /note="priming site P2"
     rep_origin      complement(1265..1653)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     CDS             complement(1758..2615)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(2616..2720)
                     /label=AmpR promoter
     terminator      2811..2897
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2989..3016
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"

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