pJLRCS vector (V005304)

Basic Vector Information

Vector Name:
pJLRCS
Antibiotic Resistance:
Ampicillin
Length:
7876 bp
Type:
Cloning vector
Replication origin:
ori
Source/Author:
Trachtmann N, Alvarez Fong KF, Guitart Font E, Sprenger GA.

pJLRCS vector Map

pJLRCS7876 bp30060090012001500180021002400270030003300360039004200450048005100540057006000630066006900720075007800flanking arm for integration5S terminatorrrnB T1 terminatorrrnB T2 terminatorAmpR promoterAmpRoribomroplacIlacIq promoterSmall ribosomal subunit protein bS20FRTcatcat promoterFRT (minimal)tac promoterlac operatoryeGFP

pJLRCS vector Sequence

LOCUS       V005304                 7876 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V005304
VERSION     V005304
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7876)
  AUTHORS   Trachtmann N, Alvarez Fong KF, Guitart Font E, Sprenger GA.
  TITLE     Construction of chromosomally encoded lacZ and gfp reporter strains
            of Escherichia coli for global regulation of metabolism
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7876)
  AUTHORS   Trachtmann N, Alvarez Fong KF, Guitart Font E, Sprenger GA.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-MAY-2016) Institute of microbiology, Stuttgart
            university, Allmandring 31, Stuttgart 70569, Germany
REFERENCE   3  (bases 1 to 7876)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7876)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (24-MAY-2016) Institute of microbiology, Stuttgart university,
            Allmandring 31, Stuttgart 70569, Germany"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7876
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    6..303
                     /label="flanking arm for integration"
                     /note="flanking arm for integration"
     regulatory      400..520
                     /label="5S terminator"
                     /note="5S terminator"
                     /regulatory_class="terminator"
     terminator      522..608
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      700..727
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        746..837
                     /label="AmpR promoter"
     CDS             838..1695
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      1869..2457
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(2643..2783)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(2888..3076)
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     CDS             complement(4097..5176)
                     /label="lacI"
                     /note="lac repressor"
     promoter        complement(5177..5254)
                     /label="lacIq promoter"
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             5605..5865
                     /gene="rpsT"
                     /label="Small ribosomal subunit protein bS20"
                     /note="Small ribosomal subunit protein bS20 from
                     Escherichia coli (strain K12). Accession#: P0A7U7"
     protein_bind    complement(5912..5959)
                     /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)."
     CDS             complement(5995..6633)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /function="chloramphenicol resistance"
                     /label="cat"
                     /protein_id="APZ86792.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRCLMNTTVLR"
     gene            complement(5995..6633)
                     /gene="cat"
                     /label="cat"
     promoter        complement(6634..6736)
                     /label="cat promoter"
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     protein_bind    complement(6842..6875)
                     /label="FRT (minimal)"
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     promoter        7067..7095
                     /label="tac promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    7103..7119
                     /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)."
     CDS             7159..7872
                     /label="yeGFP"
                     /note="yeast-enhanced green fluorescent protein"

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