LOCUS       Exported                3043 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  PCR amplification vector pCLF2, complete sequence.
ACCESSION   HM047089
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3043
                     /organism="PCR amplification vector pCLF2"
                     /mol_type="other DNA"
                     /label=derived from pCLF3 in INSD accession EU629213
                     /note="derived from pCLF3 in INSD accession EU629213"
                     /db_xref="taxon:997460"
     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"
     misc_feature    1150..1193
                     /note="FLP recognition target site; FRT"
     protein_bind    complement(1150..1183)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     regulatory      1197..1226
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     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      1265..1653
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     rep_origin      1268..1685
                     /label=R6K gamma replication origin
                     /note="R6K gamma replication origin"
     CDS             complement(1755..2615)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="confers ampicillin resistance"
                     /protein_id="ADZ54045.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(1755..2615)
                     /gene="bla"
                     /label=bla
     promoter        complement(2616..2720)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      2811..2897
                     /gene="Escherichia coli rrnB"
                     /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"