LOCUS       Exported                7104 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pNT1.1 for cKO, complete sequence.
ACCESSION   JN935771
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7104)
  AUTHORS   Tokuhiro K, Ikawa M, Benham AM, Okabe M.
  TITLE     Protein disulfide isomerase homolog PDILT is required for quality 
            control of sperm membrane protein ADAM3 and male fertility 
            [corrected]
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 109 (10), 3850-3855 (2012)
  PUBMED    22357757
REFERENCE   2  (bases 1 to 7104)
  AUTHORS   Ikawa M.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-OCT-2011) Rimd, Osaka University, 3-1 Yamada-Oka, 
            Suita, Osaka 5650871, Japan
REFERENCE   3  (bases 1 to 7104)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7104
                     /organism="Cloning vector pNT1.1 for cKO"
                     /mol_type="other DNA"
                     /db_xref="taxon:1155085"
     misc_feature    7..61
                     /note="multi-cloning site 1 (PmeI, NotI, SalI, NsiI, AsiSI,
                     XhoI, SfiI)"
     misc_feature    91..124
                     /label=LoxP site 1
                     /note="LoxP site 1"
     protein_bind    91..124
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    137..142
                     /label=BamHI
                     /note="BamHI"
     misc_feature    146..193
                     /label=FRT site 1
                     /note="FRT site 1"
     protein_bind    complement(146..193)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_feature    complement(201..1600)
                     /label=PGK-neo-pA cassette
                     /note="PGK-neo-pA cassette"
     polyA_signal    203..251
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             complement(258..1061)
                     /codon_start=1
                     /gene="aph(3)-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(1075..1574)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     misc_feature    1601..1606
                     /label=HindIII
                     /note="HindIII"
     misc_feature    1610..1657
                     /label=FRT site 2
                     /note="FRT site 2"
     protein_bind    complement(1610..1657)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_feature    1661..1686
                     /label=multi-cloning site 2 (EcoRI, NheI, AflII, AscI)
                     /note="multi-cloning site 2 (EcoRI, NheI, AflII, AscI)"
     misc_feature    1687..1720
                     /label=LoxP site 2
                     /note="LoxP site 2"
     protein_bind    1687..1720
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    1721..1750
                     /label=multi-cloning site 3 (PacI, KpnI, MfeI, AvrII)
                     /note="multi-cloning site 3 (PacI, KpnI, MfeI, AvrII)"
     misc_feature    1751..4550
                     /label=PGK-tk-pA cassette
                     /note="PGK-tk-pA cassette"
     promoter        1765..2264
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     5UTR           2306..2345
                     /label=HSV TK 5-UTR
                     /note="5 untranslated region from the herpes simplex virus
                     thymidine kinase gene"
     CDS             2363..3493
                     /codon_start=1
                     /product="herpes simplex virus thymidine kinase"
                     /label=HSV TK
                     /note="confers sensitivity to nucleoside analogs such as 
                     acyclovir or ganciclovir"
                     /translation="MASYPCHQHASAFDQAARSRGHSNRRTALRPRRQQEATEVRLEQK
                     MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWQVLGASETIANIYTTQ
                     HRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGSSHAPPPALTLIFDR
                     HPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGALPEDRHIDRLAKRQR
                     PGERLDLAMLAAIRRVYGLLANTVRYLQGGGSWREDWGQLSGTAVPPQGAEPQSNAGPR
                     PHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRPMHVFILDYDQSPAGCRDAL
                     LQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     polyA_signal    3535..3583
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     misc_feature    4558..4563
                     /label=ClaI
                     /note="ClaI"
     misc_feature    4571..7104
                     /label=SKII plasmid
                     /note="SKII plasmid"
     promoter        4927..5031
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5032..5892
                     /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      6063..6651
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6939..6960
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6975..7005
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7013..7029
                     /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     7037..7053
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7074..7092
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"