LOCUS       Exported               11474 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pAct26CRFB6, complete sequence.
ACCESSION   EF014967
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11474)
  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 11474)
  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 11474)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11474
                     /organism="Expression vector pAct26CRFB6"
                     /mol_type="other DNA"
                     /note="derived from pAct26mRFP-F; expression of zebrafish 
                     CRFB6 receptor under the control of the zebrafish 
                     beta-actin promoter"
                     /db_xref="taxon:407286"
     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             5827..6777
                     /codon_start=1
                     /product="zebrafish CRFB6 receptor"
                     /label=zebrafish CRFB6 receptor
                     /note="helical cytokine receptor; transmembrane"
                     /protein_id="ABJ97330.1"
                     /translation="MTYIIHLTVFMLIVNIERTCCLNPPQDLKIVKSQLLWKPPEVDGD
                     LRYSLQYKLDSKAEDKWYNVSSHINKNVFNITDEFYGALFRVRAEKGYHISEWAISNRV
                     NCVNVNSCAPVVNLSEKPGIVSLTLAHMDQSLEKEHGEHLEFNISSWSVNSRENPEEDV
                     VINSKDYIFNDLESGQIYCFQVEYLLYHKPYGKASKERCVFIPETPEAKKKRVLIYGPL
                     ITCFVLMVCGFCIFLFCKCKSNKRVQSVVKEWFQPFKLDLPDHYKEFLSSEFPVGLAPS
                     PSIPSLQSDDFIILKENADVEENGQEAEQQKTEIL"
     primer_bind     complement(8185..8201)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      8223..8232
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             8229..8981
                     /codon_start=1
                     /product="farnesylated mRFP1"
                     /label=farnesylated mRFP1
                     /protein_id="ABJ97331.1"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGASGLRSKLNPPDESGPGCMSCKCVLS"
     CDS             8229..8903
                     /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    9115..9236
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(9283..9301)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(9322..9338)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    9346..9362
                     /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(9370..9400)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(9724..10312)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(10483..11343)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="ABJ97332.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAISMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(11344..11448)
                     /gene="bla"
                     /label=AmpR promoter