pKC26-FB1.1 vector (V005252)

Basic Vector Information

Vector Name:
pKC26-FB1.1
Antibiotic Resistance:
Ampicillin
Length:
12615 bp
Type:
Gal4/UAS cloning vector
Replication origin:
ori
Source/Author:
Hadjieconomou D, Rotkopf S, Alexandre C, Bell DM, Dickson BJ, Salecker I.
Promoter:
hsp70

pKC26-FB1.1 vector Map

pKC26-FB1.112615 bp60012001800240030003600420048005400600066007200780084009000960010200108001140012000126005' mini-white fragmentattBM13 fwdAmpR promoterAmpRoriCAP binding sitelac promoterlac operatorM13 rev5X UAS5X UAShsp70 promoteraltered multiple cloning sitemFRT71membrane anchor; Region: Cd8aEGFPsmall t intronSV40 NLSSV40 poly(A) signalhsp70Ab polyA signalCitrineRegion: Lyn kinasemFRT71mFRT71mCD8mCherrysmall t intronSV40 NLSSV40 poly(A) signalhsp70 poly(A)Region: 1xV5 tagmCeruleanmCD8mFRT71small t intronSV40 NLSSV40 poly(A) signal

pKC26-FB1.1 vector Sequence

LOCUS       40924_26576       12615 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Gal4/UAS cloning vector pKC26-FB1.1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12615)
  AUTHORS   Hadjieconomou D, Rotkopf S, Alexandre C, Bell DM, Dickson BJ, 
            Salecker I.
  TITLE     Flybow: genetic multicolor cell labeling for neural circuit analysis
            in Drosophila melanogaster
  JOURNAL   Nat. Methods (2011) In press
  PUBMED    21297619
REFERENCE   2  (bases 1 to 12615)
  AUTHORS   Hadjieconomou D, Rotkopf S, Alexandre C, Dickson BJ, Salecker I.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-JAN-2011) Division of Molecular Neurobiology, MRC 
            National Institute for Medical Research, The Ridgeway, London, 
            England NW7 1AA, UK
REFERENCE   3  (bases 1 to 12615)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 12615)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. 
            Methods (2011) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (24-JAN-2011) Division of Molecular Neurobiology, MRC National 
            Institute for Medical Research, The Ridgeway, London, England NW7 
            1AA, UK"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..12615
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    6..866
                     /label=5' mini-white fragment
                     /note="5' mini-white fragment"
     protein_bind    884..917
                     /label=attB
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et
                     al., 2000)"
     primer_bind     complement(935..951)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1425..1529
                     /label=AmpR promoter
     CDS             1530..2387
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      2561..3149
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    3437..3458
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3473..3503
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3511..3527
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     primer_bind     3535..3551
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3598..3692
                     /label=5X UAS
                     /note="five tandem copies of the 'ScaI site' 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     protein_bind    3716..3810
                     /label=5X UAS
                     /bound_moiety="GAL4"
                     /note="five tandem copies of the ""ScaI site"" 17-mer 
                     CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) 
                     that efficiently binds yeast Gal4 (Webster et al., 1988; 
                     Pfeiffer et al., 2010)"
     promoter        3829..4067
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     misc_feature    4081..4110
                     /label=altered multiple cloning site
                     /note="altered multiple cloning site"
     misc_recomb     4111..4144
                     /label=mFRT71
                     /note="mFRT71"
     misc_feature    4151..4816
                     /note="membrane anchor; Region: Cd8a"
     CDS             4151..4816
                     /codon_start=1
                     /product="mouse lymphocyte antigen CD8 alpha chain"
                     /label=mCD8
                     /translation="MASPLTRFLSLNLLLLGESIILGSGEAKPQAPELRIFPKKMDAEL
                     GQKVDLVCEVLGSVSQGCSWLFQNSSSKLPQPTFVVYMASSHNKITWDEKLNSSKLFSA
                     MRDTNNKYVLTLNKFSKENEGYYFCSVISNSVMYFSSVVPVLQKVNSTTTKPVLRTPSP
                     VHPTGTSQPQRPEDCRPRGSVKGTGLDFACDIYIWAPLAGICVALLLSLIITLICYHSR
                     "
     CDS             4823..5539
                     /label=EGFP
                     /note="enhanced GFP"
     intron          5625..5690
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             5820..5840
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     polyA_signal    6265..6399
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    6423..6849
                     /label=hsp70 poly(A)
                     /note="polyadenlyation signal from the Drosophila
                     melanogaster hsp70Ab gene"
     regulatory      complement(6424..6852)
                     /label=hsp70Ab polyA signal
                     /note="hsp70Ab polyA signal"
                     /regulatory_class="polyA_signal_sequence"
     CDS             complement(6862..7578)
                     /label=Citrine
                     /note="enhanced variant of YFP (Heikal et al., 2001)"
     misc_feature    complement(7591..7638)
                     /label=Region: Lyn kinase
                     /note="Region: Lyn kinase"
     misc_recomb     complement(7645..7678)
                     /label=mFRT71
                     /note="mFRT71"
     misc_recomb     7691..7724
                     /label=mFRT71
                     /note="mFRT71"
     CDS             7731..8396
                     /label=mCD8
                     /note="mouse lymphocyte antigen CD8 alpha chain"
     CDS             8403..9110
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     intron          9196..9261
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             9391..9411
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     polyA_signal    9836..9970
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    complement(9978..10404)
                     /label=hsp70 poly(A)
                     /note="polyadenlyation signal from the Drosophila
                     melanogaster hsp70Ab gene"
     misc_feature    complement(10424..10468)
                     /label=Region: 1xV5 tag
                     /note="Region: 1xV5 tag"
     CDS             complement(10424..10465)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(10475..11191)
                     /label=mCerulean
                     /note="enhanced monomeric variant of CFP (Rizzo et al.,
                     2004)"
     CDS             complement(11198..11863)
                     /label=mCD8
                     /note="mouse lymphocyte antigen CD8 alpha chain"
     misc_recomb     11870..11903
                     /label=mFRT71
                     /note="mFRT71"
     intron          11993..12058
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             12188..12208
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     polyA_signal    12480..12614
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"

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