LOCUS       Exported               10914 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pAct26CRFB1, complete sequence.
ACCESSION   EF014965
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10914)
  AUTHORS   Levraud JP, Boudinot P, Colin I, Benmansour A, Peyrieras N, Herbomel
            P, Lutfalla G.
  TITLE     Identification of the zebrafish IFN receptor: implications for the 
            origin of the vertebrate IFN system
  JOURNAL   J. Immunol. 178 (7), 4385-4394 (2007)
  PUBMED    17371995
REFERENCE   2  (bases 1 to 10914)
  AUTHORS   Lutfalla G.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-SEP-2006) UMR5124, CNRS/UMII, place Eugene Bataillon, 
            Montpellier cedex 5 34095, France
REFERENCE   3  (bases 1 to 10914)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10914
                     /organism="Expression vector pAct26CRFB1"
                     /mol_type="other DNA"
                     /note="expression of zebrafish IFNLR1 (CRFB1 receptor) 
                     under the control of the zebrafish beta-actin promoter"
                     /db_xref="taxon:407284"
     rep_origin      complement(1..456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     598..614
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        624..642
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5745..7385
                     /codon_start=1
                     /product="zebrafish IFNLR1"
                     /label=zebrafish IFNLR1
                     /note="zCRFB1; helical cytokine receptor; transmembrane"
                     /protein_id="ABJ97324.1"
                     /translation="MKTSSSYTGRLFLIHLYFTVIYAIPAPVNFTIWSHNFRHILHWNY
                     GVNSPPQSVFNLQRFEKGKQTEHLNIRNTTMDVSEACQDIYIPCTFLIWASLDNMNSSV
                     IEKKFIPYEDTIIGPPVIFLSGCGDCLNISISLPNESRKDDKLRRFYNSVSYSISWKKH
                     GNNEVREILRSQEPSKQYVLENLQPGVQYCVKVLPQINSNQNTQPSSWQCEYTSKEEAQ
                     RVLYFMSWLLGATLSGSCVMMLAWILVYTGFLCKPKNPPLKSLSNIVPATYLIPEQTLS
                     ESLSLTEVQLIHLSYDTTENNSQLKNPEKREYEGSVLKSDTEDEEDSNDEDEVKCVYMS
                     CEIDDISCQNNSTKKDIVSALPFNKSSIVCYTKEPAMEKIQYDDLKSEKELLSELFEEG
                     HGHLEVTNTQLKTEGNEKGSTNTPGNINLFSVTVRTFGAEDDLDKEDVYKPLLPKLFVK
                     TDKWTEELLPVSNKIQISIPIETGMKTDDLIHLDSQNDLVHVCSLAHNVETNPDTTMSA
                     YMMNHIGKMHDQNKDLLEETNCDTSYITR"
     primer_bind     complement(7625..7641)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      7663..7672
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             7669..8421
                     /codon_start=1
                     /product="farnesylated mRFP1"
                     /label=farnesylated mRFP1
                     /protein_id="ABJ97325.1"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGASGLRSKLNPPDESGPGCMSCKCVLS"
     CDS             7669..8343
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     polyA_signal    8555..8676
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(8723..8741)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8762..8778)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8786..8802
                     /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(8810..8840)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(9164..9752)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(9923..10783)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="ABJ97326.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAISMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(10784..10888)
                     /gene="bla"
                     /label=AmpR promoter