LOCUS       Exported                8416 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector krt4-sfGFP-tVE1, complete sequence.
ACCESSION   KU645307
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8416)
  AUTHORS   Eskova A, Garcia-Junco R, Chauvigne F, Maischein H-M., Ammelburg M, 
            Cerda J, Kaderali L, Nusslein-Volhard C, Irion U.
  TITLE     Gain-of-function mutations of mau/DrAqp3a influence zebrafish 
            pigment pattern formation through the tissue environment
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8416)
  AUTHORS   Eskova A.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-JAN-2016) e-CNV group, Max Planck Institute for 
            Developmental Biology, Spemannstr. 35-39, Tuebingen 72076, Germany
REFERENCE   3  (bases 1 to 8416)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8416
                     /organism="Cloning vector krt4-sfGFP-tVE1"
                     /mol_type="other DNA"
                     /label=derived from Danio rerio
                     /note="derived from Danio rerio"
                     /db_xref="taxon:1891373"
     primer_bind     1061..1077
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1097..1117
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        1119..1137
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1163..1179
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      1195..3433
                     /regulatory_class="promoter"
                     /note="krt4"
     regulatory      3455..3464
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3461..5083
                     /codon_start=1
                     /product="SfGFP/Aqp3aR220Q fusion protein"
                     /label=SfGFP/Aqp3aR220Q fusion protein
                     /note="tVE1 allele"
                     /protein_id="AOE47049.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKSGLRSMGWQKSVLDKLAQTFQIRNKLLRQGLAECLGTLILVM
                     FGCGSLAQLKLSEGSHGLFLTANLAFGFGATLGILVCGQVSGGHLNPAVTFALCLLGRE
                     KWRKFPVYFLFQTLGSFLGAAIIFAEYHDAIYDYAGESNELLVLGEKETAGIFATYPSK
                     YLTPLNGFFDQVIGTASLIVCILAIVDPYNNPIPQGLEAFTVGFSVLIIGLSMGFNSGY
                     AVNPAQDFGPRLFTAMAGWGSEVFTARDYWFLVPIFAPFIGAVIGVIVYQLMVGWHVEG
                     EARDKKAKAREEVMNLNDVASKE"
     polyA_signal    5116..5250
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(5313..5333)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(5341..5357)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    5373..5507
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6179..6195)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(6232..6250)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6271..6287)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6295..6311
                     /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)."
     promoter        complement(6319..6349)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6364..6385
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6673..7261)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7432..8292)
                     /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"
     promoter        complement(8293..8397)
                     /gene="bla"
                     /label=AmpR promoter