LOCUS       Exported               10550 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pI2zCXCR4b6.6kbprom_mCherryF, complete sequence.
ACCESSION   GU394077
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10550)
  AUTHORS   Gamba L, Cubedo N, Ghysen A, Lutfalla G, Dambly-Chaudiere C.
  TITLE     Estrogen receptor ESR1 controls cell migration by repressing 
            chemokine receptor CXCR4 in the zebrafish posterior lateral line 
            system
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 107 (14), 6358-6363 (2010)
  PUBMED    20308561
REFERENCE   2  (bases 1 to 10550)
  AUTHORS   Gamba L, Cubedo N, Ghysen A, Lutfalla G, Dambly-Chaudiere C.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-JAN-2010) DIMNP, CNRS/Universite Montpellier2, Place 
            Eugene Bataillon, Montpellier, Cedex5 F-34095, France
REFERENCE   3  (bases 1 to 10550)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10550
                     /organism="Expression vector pI2zCXCR4b6.6kbprom_mCherryF"
                     /mol_type="other DNA"
                     /db_xref="taxon:754156"
     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"
     misc_feature    656..673
                     /label=I-SceI meganuclease site
                     /note="I-SceI meganuclease site"
     gene            695..7261
                     /gene="CXCR4b"
                     /label=CXCR4b
                     /note="derived from zebrafish"
     misc_feature    695..6647
                     /gene="CXCR4b"
                     /note="5 flanking sequence; derived from zebrafish"
     CDS             complement(5657..5668)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      6647..6791
                     /regulatory_class="promoter"
                     /gene="CXCR4b"
                     /note="derived from zebrafish"
     regulatory      6745..6791
                     /regulatory_class="promoter"
                     /gene="CXCR4b"
                     /note="minimal promoter; derived from zebrafish"
     exon            6792..6895
                     /gene="CXCR4b"
                     /number=1
                     /note="derived from zebrafish"
     misc_difference 6881
                     /gene="CXCR4b"
                     /replace="a"
                     /label=ATG start codon mutated to GTG
                     /note="ATG start codon mutated to GTG"
     intron          6896..7247
                     /gene="CXCR4b"
                     /number=1
                     /note="derived from zebrafish"
     exon            7248..7261
                     /gene="CXCR4b"
                     /number=2
                     /note="derived from zebrafish"
     regulatory      7284..7293
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             7290..8057
                     /codon_start=1
                     /product="mCherry-F"
                     /label=mCherry-F
                     /note="farnesylated mCherry version of the monomeric red 
                     fluorescent protein"
                     /protein_id="ADF31907.1"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGASGLRSKLNPPDESGPGCMSCKCVLS"
     regulatory      8100..8300
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV 40 polyA signal"
     polyA_signal    8191..8312
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    8323..8340
                     /label=I-SceI meganuclease site
                     /note="I-SceI meganuclease site"
     misc_feature    8341..655
                     /label=Bluescript SK (+)
                     /note="Bluescript SK (+)"
     promoter        complement(8359..8377)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8398..8414)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8422..8438
                     /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(8446..8476)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(8800..9388)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(9559..10419)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAISMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(10420..10524)
                     /gene="bla"
                     /label=AmpR promoter