pCFPbglS vector (V008553)

Basic Vector Information

Vector Name:
pCFPbglS
Antibiotic Resistance:
Ampicillin
Length:
6032 bp
Type:
Cloning vector
Replication origin:
ori
Source/Author:
Bisicchia P, Botella E, Devine KM.
Promoter:
T3

pCFPbglS vector Map

pCFPbglS6032 bp30060090012001500180021002400270030003300360039004200450048005100540057006000f1 oriM13 fwdT7 promoterSK primerbglS frontLIC sitemECFPKanRKS primerbglS backT3 promoterM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoter

pCFPbglS vector Sequence

LOCUS       40924_10511        6032 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCFPbglS, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6032)
  AUTHORS   Bisicchia P, Botella E, Devine KM.
  TITLE     Suite of novel vectors for ectopic insertion of GFP, CFP and IYFP 
            transcriptional fusions in single copy at the amyE and bglS loci in 
            Bacillus subtilis
  JOURNAL   Plasmid 64 (3), 143-149 (2010)
  PUBMED    20600285
REFERENCE   2  (bases 1 to 6032)
  AUTHORS   Bisicchia P.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-MAY-2010) Trinity College Dublin, 13 Meades Terrace, 
            Dublin 2, Ireland
REFERENCE   3  (bases 1 to 6032)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6032)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid"; 
            date: "2010"; volume: "64"; issue: "3"; pages: "143-149"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-MAY-2010) Trinity College Dublin, 13 Meades Terrace, Dublin 2, 
            Ireland"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6032
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(6..461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     677..693
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_recomb     695..1121
                     /label=bglS front
                     /note="bglS front"
     misc_feature    1134..1169
                     /label=LIC site
                     /note="LIC site"
     CDS             1170..1886
                     /codon_start=1
                     /label=mECFP
                     /note="enhanced CFP with monomerizing A206K mutation
                     (Zacharias et al., 2002)"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             2446..3237
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     primer_bind     complement(3397..3413)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_recomb     3416..3813
                     /label=bglS back
                     /note="bglS back"
     promoter        complement(3847..3865)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3886..3902)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3910..3926)
                     /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)."
     promoter        complement(3934..3964)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3979..4000)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4288..4876)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5050..5907)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5908..6012)
                     /label=AmpR promoter

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