pKC26-FB1.0 vector (V005253)

LOCUS       40924_26571        9041 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Gal4/UAS cloning vector pKC26-FB1.0, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9041)
  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 9041)
  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 9041)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9041)
  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..9041
                     /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
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     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
                     /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..4116
                     /label=multiple cloning site
                     /note="multiple cloning site"
     misc_recomb     4117..4150
                     /label=mFRT71
                     /note="mFRT71"
     CDS             4157..4822
                     /codon_start=1
                     /label=mCD8
                     /note="mouse lymphocyte antigen CD8 alpha chain"
                     /translation="MASPLTRFLSLNLLLLGESIILGSGEAKPQAPELRIFPKKMDAEL
                     GQKVDLVCEVLGSVSQGCSWLFQNSSSKLPQPTFVVYMASSHNKITWDEKLNSSKLFSA
                     MRDTNNKYVLTLNKFSKENEGYYFCSVISNSVMYFSSVVPVLQKVNSTTTKPVLRTPSP
                     VHPTGTSQPQRPEDCRPRGSVKGTGLDFACDIYIWAPLAGICVALLLSLIITLICYHSR
                     "
     CDS             4829..5536
                     /codon_start=1
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     intron          5622..5687
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             5817..5837
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     polyA_signal    6262..6396
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    complement(6404..6830)
                     /label=hsp70 poly(A)
                     /note="polyadenlyation signal from the Drosophila
                     melanogaster hsp70Ab gene"
     misc_feature    complement(6850..6894)
                     /label=Region: 1xV5 tag
                     /note="Region: 1xV5 tag"
     CDS             complement(6850..6891)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             complement(6901..7617)
                     /codon_start=1
                     /label=mCerulean
                     /note="enhanced monomeric variant of CFP (Rizzo et al.,
                     2004)"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLNL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(7624..8289)
                     /codon_start=1
                     /label=mCD8
                     /note="mouse lymphocyte antigen CD8 alpha chain"
                     /translation="MASPLTRFLSLNLLLLGESIILGSGEAKPQAPELRIFPKKMDAEL
                     GQKVDLVCEVLGSVSQGCSWLFQNSSSKLPQPTFVVYMASSHNKITWDEKLNSSKLFSA
                     MRDTNNKYVLTLNKFSKENEGYYFCSVISNSVMYFSSVVPVLQKVNSTTTKPVLRTPSP
                     VHPTGTSQPQRPEDCRPRGSVKGTGLDFACDIYIWAPLAGICVALLLSLIITLICYHSR
                     "
     misc_recomb     complement(8296..8329)
                     /label=mFRT71
                     /note="mFRT71"
     intron          8419..8484
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             8614..8634
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     polyA_signal    8906..9040
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"