LOCUS       Exported               10504 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Knock-in vector FBDki, complete sequence.
ACCESSION   HM357235
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10504)
  AUTHORS   Tamayev R, Giliberto L, Matsuda S, DAbramo C, Arancio O, Vidal R, 
            DAdamio L.
  TITLE     Memory deficits due to Familial British Dementia BRI2 mutation are 
            caused by loss of BRI2 function rather than amyloidosis
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10504)
  AUTHORS   Giliberto L, DAdamio L.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-MAY-2010) Litwin-Zucker Center for the Study of 
            Alzheimers Disease, The Feinstein Institute for Medical 
            Research-North Shore-LIJ, 350 Community Drive, Manhasset, NY 11030, 
            USA
REFERENCE   3  (bases 1 to 10504)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10504
                     /organism="Knock-in vector FBDki"
                     /mol_type="other DNA"
                     /PCR_primers="fwd_name: BAC Primer, fwd_seq: 
                     cccaagctttttttttttttttttaaagacaac, rev_name: BAC Primer, 
                     rev_seq: gggaagcttgaagtggtcagcagggag"
                     /PCR_primers="fwd_name: ES Cells-Left Arm Genomic Primer,
                     fwd_seq: cagtcctaccttatccatgagcac, rev_name: ES Cells-Left 
                     Arm Neo Cassette Primer, rev_seq: cttcctcgtgctttacggtatc"
                     /PCR_primers="fwd_name: ES Cells-Right Arm Neo Cassette
                     Primer, fwd_seq: tgcacgagactagtgagacgtg, rev_name: ES 
                     Cells-Right Arm Genomic Primer, rev_seq: 
                     agtctgtatctcactacggcatcc"
                     /PCR_primers="fwd_name: Genomic Upstream Primer for PCR
                     and Digestion, fwd_seq: cagtcctaccttatccatgagcac, rev_name:
                     Genomic Downstream Primer for PCR and Digestion, rev_seq: 
                     agtctgtatctcactacggcatcc"
                     /PCR_primers="fwd_name: Mice-Targeting Vector-Genomic
                     Set1, fwd_seq: atggcaccacccacaatagg, rev_name: 
                     Mice-Targeting Vector-Genomic Set1/2, rev_seq: 
                     cctagcaactggtaacagtgc, rev_seq: ggtaagctctaaggagaagcg"
                     /note="artificially derived by cloning part of the genomic 
                     sequence of the ITMB2 mouse gene into a Soriano vector 
                     containing DTA and neomycin resistance cassette, flanked by
                     LoxPs; familial british dementia knock-in"
                     /db_xref="taxon:908007"
     misc_feature    complement(1..2221)
                     /label=Soriano vector
                     /note="Soriano vector"
     promoter        24..128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             129..989
                     /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"
     rep_origin      1160..1748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2036..2057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2072..2102
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2110..2126
                     /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)."
     primer_bind     2134..2150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2171..2189
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      complement(2223..2693)
                     /regulatory_class="polyA_signal_sequence"
     polyA_signal    2257..2481
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     intron          2555..2620
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             complement(2695..3267)
                     /codon_start=1
                     /gene="Corynebacterium diphtheriae tox"
                     /product="diphtheria toxin fragment A"
                     /label=DTA
                     /translation="DDVVDSSKSFVMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNYD
                     DDWKGFYSTDNKYDAAGYSVDNENPLSGKAGDVVKVTYPGLTKVLALKVDNAETIKKEL
                     GLSLTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVKLEI
                     NFETRGKRGQDAMYEYMAQACAGNRVRR"
     CDS             complement(3072..3131)
                     /codon_start=1
                     /product="DTA toxin"
                     /label=DTA toxin
                     /protein_id="ADO40088.1"
                     /translation="MIGKGFIVPTINTTLRDTL"
     regulatory      complement(3286..3797)
                     /regulatory_class="promoter"
                     /note="PGK1 promoter"
     promoter        complement(3294..3788)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     misc_feature    3828..4989
                     /note="left arm; contains intron 5"
     misc_feature    5038..5072
                     /label=LoxP
                     /note="LoxP"
     protein_bind    5038..5071
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     polyA_signal    5130..5354
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(5392..6195)
                     /codon_start=1
                     /product="neomycin resistance protein"
                     /label=neomycin resistance protein
                     /protein_id="ADO40089.1"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     regulatory      complement(6201..6712)
                     /regulatory_class="promoter"
                     /note="PGK1 promoter"
     promoter        complement(6209..6703)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     misc_feature    6733..6767
                     /label=LoxP
                     /note="LoxP"
     protein_bind    6733..6766
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     primer_bind     complement(6780..6796)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    6805..6825
                     /label=pBlueScript MCS fragment
                     /note="pBlueScript MCS fragment"
     primer_bind     6805..6821
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    6829..9835
                     /note="right arm fragment; contains intron 5, exon 6, and 
                     intron 6"
     exon            7037..7156
                     /number=6
     CDS             7037..7156
                     /codon_start=1
                     /product="ITM2B integral membrane protein 2B"
                     /label=ITM2B integral membrane protein 2B
                     /note="ABRI"
                     /protein_id="ADO40090.1"
                     /translation="GIQKREASNCFAIRHFENKFAVETLICSRTVKKNIIEEN"
     misc_difference 7070
                     /replace="a"
                     /label=humanized
                     /note="humanized"
     misc_feature    7121..7156
                     /label=ABRI mutant-extra bases
                     /note="ABRI mutant-extra bases"
     misc_difference 7121
                     /replace="t"
                     /label=ABRI mutation
                     /note="ABRI mutation"
     3UTR           7157..7960
                     /label=ITM2B 3 UTR
                     /note="ITM2B 3 UTR"
     misc_feature    complement(9836..10504)
                     /label=Soriano vector
                     /note="Soriano vector"
     promoter        complement(9861..9879)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9889..9905)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      10047..10502
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"