LOCUS       Exported                7208 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSINTK DNA, complete sequence.
ACCESSION   AB242435
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7208)
  AUTHORS   Suemizu H, Ito M.
  TITLE     pSINTK cloning vector for homologous recombination
  JOURNAL   Published Only in Database (2005)
REFERENCE   2  (bases 1 to 7208)
  AUTHORS   Suemizu H, Ito M.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-NOV-2005) Contact:Hiroshi Suemizu Central Institute 
            for Experimental Animals, Biomedical Research Department; 3-25-12 
            Tonomachi, Kawasaki-ku, Kawasaki, Kanagawa 210-0821, Japan
REFERENCE   3  (bases 1 to 7208)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7208
                     /organism="Cloning vector pSINTK"
                     /mol_type="other DNA"
                     /db_xref="taxon:357972"
     promoter        36..54
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    72..96
                     /note="multiple cloning site 1 (Sfi I, Not I, Nhe I, Kpn 
                     I)"
     misc_feature    141..181
                     /label=lox p site 1
                     /note="lox p site 1"
     protein_bind    complement(148..181)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    366..1994
                     /label=neomycin resistance gene expression cassette
                     /note="neomycin resistance gene expression cassette"
     regulatory      366..881
                     /regulatory_class="promoter"
                     /gene="mouse phosphoglycerate kinase-1"
     gene            366..881
                     /gene="mouse phosphoglycerate kinase-1"
                     /label=mouse phosphoglycerate kinase-1
     promoter        378..877
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             891..1694
                     /codon_start=1
                     /gene="neo"
                     /product="neomycin resistance protein"
                     /label=neo
                     /protein_id="BAE48697.1"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            891..1694
                     /gene="neo"
                     /label=neo
     misc_feature    1703..1884
                     /label=DNA fragment contains poly A signal
                     /note="DNA fragment contains poly A signal"
     regulatory      1738..1743
                     /regulatory_class="polyA_signal_sequence"
                     /note="poly A signal"
     misc_feature    2007..2041
                     /label=lox p site 2
                     /note="lox p site 2"
     protein_bind    complement(2008..2041)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    2055..2088
                     /label=multiple cloning site 2 (Xba I, Eco T22I, Xho I)
                     /note="multiple cloning site 2 (Xba I, Eco T22I, Xho I)"
     misc_feature    2089..4413
                     /label=HSV-TK gene expression cassette
                     /note="HSV-TK gene expression cassette"
     regulatory      2089..2596
                     /regulatory_class="promoter"
                     /gene="mouse phosphoglycerate kinase-1"
     gene            2089..2596
                     /gene="mouse phosphoglycerate kinase-1"
                     /label=mouse phosphoglycerate kinase-1
     promoter        2093..2592
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     5UTR           2634..2673
                     /label=HSV TK 5-UTR
                     /note="5 untranslated region from the herpes simplex virus
                     thymidine kinase gene"
     CDS             2691..3821
                     /codon_start=1
                     /gene="HSV-TK"
                     /product="thymidine kinase"
                     /label=HSV-TK
                     /protein_id="BAE48698.1"
                     /translation="MASYPCHQHASAFDQAARSRGHSNRRTALRPRRQQEATEVRLEQK
                     MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWQVLGASETIANIYTTQ
                     HRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGSSHAPPPALTLIFDR
                     HPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGALPEDRHIDRLAKRQR
                     PGERLDLAMLAAIRRVYGLLANTVRYLQGGGSWREDWGQLSGTAVPPQGAEPQSNAGPR
                     PHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRPMHVFILDYDQSPAGCRDAL
                     LQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     gene            2691..3821
                     /gene="HSV-TK"
                     /label=HSV-TK
     misc_feature    3822..4413
                     /label=DNA fragment contains poly A signal
                     /note="DNA fragment contains poly A signal"
     polyA_signal    3863..3911
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     regulatory      3880..3885
                     /regulatory_class="polyA_signal_sequence"
                     /note="poly A signal"
     promoter        complement(4430..4448)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4458..4474)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4616..5071
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5097..5201
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5202..6062
                     /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      6233..6821
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    7109..7130
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7145..7175
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7183..7199
                     /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     7207..15
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"