LOCUS       Exported                6465 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector mTol2-B-P-GFF-CR3, complete sequence.
ACCESSION   KC800699
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6465)
  AUTHORS   Otsuna H, Scoresby AN, Gaynes B, Tong ZZ, Kwan KK, Hutcheson DA, 
            Chien CB, Dorsky R.
  TITLE     Large Scale Zebrafish Gal4 Enhancer Trap Screen for Neural Specific 
            Expression Patterns
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6465)
  AUTHORS   Otsuna H, Scoresby AN, Gaynes B, Tong ZZ, Kwan KK, Hutcheson DA, 
            Chien CB, Dorsky R.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-MAR-2013) Neurobiology and Anatomy, University of 
            Utah, 20 North 1900 East, Salt Lake City, UT 84132, USA
REFERENCE   3  (bases 1 to 6465)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6465
                     /organism="Expression vector mTol2-B-P-GFF-CR3"
                     /mol_type="other DNA"
                     /db_xref="taxon:1344358"
     primer_bind     complement(24..40)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(25..40)
                     /label=M13F
                     /note="M13F"
     rep_origin      181..636
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      198..504
                     /label=f1(+) ori
                     /note="f1(+) ori"
     promoter        714..818
                     /gene="bla"
                     /label=AmpR promoter
     CDS             819..1679
                     /codon_start=1
                     /product="ampR"
                     /label=ampR
                     /protein_id="AGN90800.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1774..2456
                     /label=colE1 ori
                     /note="colE1 ori"
     rep_origin      1824..2497
                     /label=pUC ori
                     /note="pUC ori"
     rep_origin      1850..2438
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2726..2747
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2762..2792
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2800..2816
                     /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     2824..2845
                     /label=M13R
                     /note="M13R"
     primer_bind     2824..2840
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2858..2876
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     regulatory      2858..2874
                     /regulatory_class="promoter"
                     /note="SP6"
     repeat_region   2927..2943
                     /label=Tol2 3 TIR
                     /note="Tol2 3 TIR"
     primer_bind     3108..3129
                     /label=M13R
                     /note="M13R"
     primer_bind     3108..3124
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_recomb     3144..3164
                     /label=attB4
                     /note="attB4"
     misc_feature    3144..3164
                     /label=att4_shared
                     /note="att4_shared"
     protein_bind    3144..3164
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     regulatory      3193..3203
                     /regulatory_class="promoter"
                     /note="adenovirus E1B TATA"
     5UTR           3211..3242
                     /label=carp beta-actin trx start
                     /note="carp beta-actin trx start"
     protein_bind    3248..3272
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_recomb     3249..3272
                     /label=attB1
                     /note="attB1"
     misc_recomb     3249..3255
                     /label=attB1
                     /note="attB1"
     misc_feature    3253..3266
                     /label=att1_shared
                     /note="att1_shared"
     misc_feature    3253..3255
                     /label=att1_shared
                     /note="att1_shared"
     misc_recomb     3256..3272
                     /label=attL1
                     /note="attL1"
     misc_feature    3256..3266
                     /label=att1_shared
                     /note="att1_shared"
     misc_feature    3281..3820
                     /label=Gal4FF
                     /note="Gal4FF"
     CDS             3281..3721
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     misc_feature    3281..3720
                     /label=Gal4 DBD
                     /note="Gal4 DBD"
     protein_bind    complement(3822..3846)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_recomb     3822..3845
                     /label=attB2
                     /note="attB2"
     misc_recomb     3822..3843
                     /label=attL2
                     /note="attL2"
     misc_feature    3828..3843
                     /label=att2_shared
                     /note="att2_shared"
     misc_recomb     3844..3845
                     /label=attB2
                     /note="attB2"
     misc_feature    3848..4043
                     /label=SV40 late polyA (from pCS2+)
                     /note="SV40 late polyA (from pCS2+)"
     polyA_signal    3848..3982
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_recomb     4045..4065
                     /label=attB3
                     /note="attB3"
     protein_bind    complement(4045..4065)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     misc_feature    4045..4059
                     /label=att3_shared
                     /note="att3_shared"
     primer_bind     complement(4073..4089)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4074..4089)
                     /label=M13F
                     /note="M13F"
     misc_feature    4105..4302
                     /label=SV40 late polyA from pCS2+
                     /note="SV40 late polyA from pCS2+"
     polyA_signal    4105..4239
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    complement(4366..4563)
                     /label=SV40 late polyA from pCS2+
                     /note="SV40 late polyA from pCS2+"
     polyA_signal    4429..4563
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        4568..4585
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(4601..5314)
                     /codon_start=1
                     /product="TagRFP"
                     /label=TagRFP
                     /protein_id="AGN90799.1"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     regulatory      complement(5315..6213)
                     /regulatory_class="promoter"
                     /note="cmlc2 promoter"
     repeat_region   6433..6451
                     /label=Tol2 5 TIR
                     /note="Tol2 5 TIR"
     primer_bind     complement(6461..18)
                     /label=T7 primer
                     /note="T7 primer"
     promoter        complement(6464..17)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"