LOCUS       Exported               12615 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Gal4/UAS cloning vector pKC26-FB1.1, complete sequence.
ACCESSION   HQ998854
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12615
                     /organism="Gal4/UAS cloning vector pKC26-FB1.1"
                     /mol_type="other DNA"
                     /db_xref="taxon:983950"
     misc_feature    6..866
                     /label=5 mini-white fragment
                     /note="5 mini-white fragment"
     misc_recomb     873..925
                     /label=attB site
                     /note="attB site"
     protein_bind    884..917
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /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
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1530..2390
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2561..3149
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    3437..3458
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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"
     misc_feature    3598..3808
                     /label=10xUAS
                     /note="10xUAS"
     protein_bind    3598..3692
                     /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)"
     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"
     CDS             4151..5542
                     /codon_start=1
                     /product="Cd8a-EGFP fusion protein"
                     /label=Cd8a-EGFP fusion protein
                     /protein_id="ADY04825.1"
                     /translation="MASPLTRFLSLNLLLLGESIILGSGEAKPQAPELRIFPKKMDAEL
                     GQKVDLVCEVLGSVSQGCSWLFQNSSSKLPQPTFVVYMASSHNKITWDEKLNSSKLFSA
                     MRDTNNKYVLTLNKFSKENEGYYFCSVISNSVMYFSSVVPVLQKVNSTTTKPVLRTPSP
                     VHPTGTSQPQRPEDCRPRGSVKGTGLDFACDIYIWAPLAGICVALLLSLIITLICYHSR
                     GSMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVP
                     WPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFE
                     GDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDG
                     SVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGM
                     DELYK"
     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..5542
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    4823..5539
                     /label=Region: EGFP
                     /note="Region: EGFP"
     regulatory      5549..6399
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 polyA signal"
     intron          5625..5690
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             5820..5840
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     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)
                     /regulatory_class="polyA_signal_sequence"
                     /note="hsp70Ab polyA signal"
     CDS             complement(6859..7638)
                     /codon_start=1
                     /product="membrane-anchored mCitrine protein"
                     /label=membrane-anchored mCitrine protein
                     /note="contains Lyn kinase palm/myr signal membrane anchor"
                     /protein_id="ADY04826.1"
                     /translation="MGCIKSKRKDNLNDDEAAGSMVSKGEELFTGVVPILVELDGDVNG
                     HKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDF
                     FKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLE
                     YNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYL
                     SYQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     misc_feature    complement(6862..7578)
                     /label=Region: mCitrine
                     /note="Region: mCitrine"
     CDS             complement(6862..7578)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     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..9113
                     /codon_start=1
                     /product="Cd8a-mCherry fusion protein"
                     /label=Cd8a-mCherry fusion protein
                     /protein_id="ADY04827.1"
                     /translation="MASPLTRFLSLNLLLLGESIILGSGEAKPQAPELRIFPKKMDAEL
                     GQKVDLVCEVLGSVSQGCSWLFQNSSSKLPQPTFVVYMASSHNKITWDEKLNSSKLFSA
                     MRDTNNKYVLTLNKFSKENEGYYFCSVISNSVMYFSSVVPVLQKVNSTTTKPVLRTPSP
                     VHPTGTSQPQRPEDCRPRGSVKGTGLDFACDIYIWAPLAGICVALLLSLIITLICYHSR
                     GSMVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGTQTAKLKVTKGG
                     PLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVVTVTQDS
                     SLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGEIKQRLKLKDG
                     GHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAEGRHSTGGMDEL
                     YK"
     misc_feature    7731..8396
                     /note="membrane anchor; Region: Cd8a"
     CDS             7731..8396
                     /codon_start=1
                     /product="mouse lymphocyte antigen CD8 alpha chain"
                     /label=mCD8
                     /translation="MASPLTRFLSLNLLLLGESIILGSGEAKPQAPELRIFPKKMDAEL
                     GQKVDLVCEVLGSVSQGCSWLFQNSSSKLPQPTFVVYMASSHNKITWDEKLNSSKLFSA
                     MRDTNNKYVLTLNKFSKENEGYYFCSVISNSVMYFSSVVPVLQKVNSTTTKPVLRTPSP
                     VHPTGTSQPQRPEDCRPRGSVKGTGLDFACDIYIWAPLAGICVALLLSLIITLICYHSR
                     "
     CDS             8403..9113
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     misc_feature    8403..9110
                     /label=Region: mCherry
                     /note="Region: mCherry"
     regulatory      9120..9970
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 polyA signal"
     intron          9196..9261
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             9391..9411
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    9836..9970
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    9978..10404
                     /label=hsp70 poly(A)
                     /note="polyadenlyation signal from the Drosophila 
                     melanogaster hsp70Ab gene"
     regulatory      complement(9979..10407)
                     /regulatory_class="polyA_signal_sequence"
                     /note="hsp70Ab polyA signal"
     CDS             complement(10409..11863)
                     /codon_start=1
                     /product="Cd8a-Cerulean-1xV5 tag fusion protein"
                     /label=Cd8a-Cerulean-1xV5 tag fusion protein
                     /protein_id="ADY04828.1"
                     /translation="MASPLTRFLSLNLLLLGESIILGSGEAKPQAPELRIFPKKMDAEL
                     GQKVDLVCEVLGSVSQGCSWLFQNSSSKLPQPTFVVYMASSHNKITWDEKLNSSKLFSA
                     MRDTNNKYVLTLNKFSKENEGYYFCSVISNSVMYFSSVVPVLQKVNSTTTKPVLRTPSP
                     VHPTGTSQPQRPEDCRPRGSVKGTGLDFACDIYIWAPLAGICVALLLSLIITLICYHSR
                     GSMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLNLKFICTTGKLPVP
                     WPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFE
                     GDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIKANFKIRHNIEDG
                     SVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLEFVTAAGITLGM
                     DELYKSPSGKPIPNPLLGLDSTAGLT"
     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"
     misc_feature    complement(10472..11191)
                     /label=Region: Cerulean
                     /note="Region: Cerulean"
     CDS             complement(10475..11191)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLNL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    complement(11198..11863)
                     /note="membrane anchor; Region: Cd8a"
     CDS             complement(11198..11863)
                     /codon_start=1
                     /product="mouse lymphocyte antigen CD8 alpha chain"
                     /label=mCD8
                     /translation="MASPLTRFLSLNLLLLGESIILGSGEAKPQAPELRIFPKKMDAEL
                     GQKVDLVCEVLGSVSQGCSWLFQNSSSKLPQPTFVVYMASSHNKITWDEKLNSSKLFSA
                     MRDTNNKYVLTLNKFSKENEGYYFCSVISNSVMYFSSVVPVLQKVNSTTTKPVLRTPSP
                     VHPTGTSQPQRPEDCRPRGSVKGTGLDFACDIYIWAPLAGICVALLLSLIITLICYHSR
                     "
     misc_recomb     11870..11903
                     /label=mFRT71
                     /note="mFRT71"
     regulatory      11917..12615
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 polyA signal"
     intron          11993..12058
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             12188..12208
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    12480..12614
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"