LOCUS       Exported                9494 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector eif1a-dsRed-eif1a-sfGFP-tVE1, complete sequence.
ACCESSION   KU645306
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9494)
  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  JOURNAL   Unpublished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 9494)
  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 9494)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9494
                     /organism="Cloning vector eif1a-dsRed-eif1a-sfGFP-tVE1"
                     /mol_type="other DNA"
                     /label=derived from Danio rerio
                     /note="derived from Danio rerio"
                     /db_xref="taxon:1891372"
     primer_bind     111..127
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      438..1597
                     /regulatory_class="promoter"
                     /note="eif1alpha"
     intron          959..1531
                     /label=-beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     protein_bind    1600..1633
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      1640..1649
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1646..2323
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     CDS             1679..2323
                     /codon_start=1
                     /product="DsRed"
                     /label=DsRed
                     /note="red fluorescent protein"
                     /protein_id="AOE47047.1"
                     /translation="MRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVKLKVTKGGPLPF
                     AWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGVATVTQDSSLQD
                     GCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHKALKLKDGGHYL
                     VEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHHLFL"
     polyA_signal    2444..2565
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    2819..2852
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      2900..4059
                     /regulatory_class="promoter"
                     /note="eif1alpha"
     intron          3421..3993
                     /label=-beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     5UTR           4068..4110
                     /label=Xenopus globin 5-UTR
                     /note="translational enhancer from the 5-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     regulatory      4135..4144
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4141..5763
                     /codon_start=1
                     /product="SfGFP/Aqp3aR220Q fusion protein"
                     /label=SfGFP/Aqp3aR220Q fusion protein
                     /note="tVE1 allele"
                     /protein_id="AOE47048.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKSGLRSMGWQKSVLDKLAQTFQIRNKLLRQGLAECLGTLILVM
                     FGCGSLAQLKLSEGSHGLFLTANLAFGFGATLGILVCGQVSGGHLNPAVTFALCLLGRE
                     KWRKFPVYFLFQTLGSFLGAAIIFAEYHDAIYDYAGESNELLVLGEKETAGIFATYPSK
                     YLTPLNGFFDQVIGTASLIVCILAIVDPYNNPIPQGLEAFTVGFSVLIIGLSMGFNSGY
                     AVNPAQDFGPRLFTAMAGWGSEVFTARDYWFLVPIFAPFIGAVIGVIVYQLMVGWHVEG
                     EARDKKAKAREEVMNLNDVASKE"
     polyA_signal    5796..5930
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(5993..6013)
                     /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(6021..6037)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    6053..6187
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(6769..6787)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6805..6821)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6829..6845
                     /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(6853..6883)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6898..6919
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(7207..7795)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7966..8826)
                     /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(8827..8931)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(9009..9464)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"