LOCUS       Exported                4570 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pBlueTAL, complete sequence.
ACCESSION   KF724948
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4570)
  AUTHORS   Takasu Y, Sajwan S, Daimon T, Osanai-Futahashi M, Uchino K, Sezutsu 
            H, Tamura T, Zurovec M.
  TITLE     Efficient TALEN construction for Bombyx mori gene targeting
  JOURNAL   PLoS ONE 8 (9), E73458 (2013)
  PUBMED    24058473
REFERENCE   2  (bases 1 to 4570)
  AUTHORS   Takasu Y, Sajwan S, Daimon T, Osanai-Futahashi M, Uchino K, Sezutsu 
            H, Tamura T, Zurovec M.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-OCT-2013) Genetics, Biology Centre, CAS, Institute of 
            Entomology, Branisovska 31, Ceske Budejovice 37005, Czech Republic
REFERENCE   3  (bases 1 to 4570)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4570
                     /organism="Cloning vector pBlueTAL"
                     /mol_type="other DNA"
                     /db_xref="taxon:1435192"
     rep_origin      complement(6..461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        542..560
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      542..559
                     /regulatory_class="promoter"
                     /note="T7 promoter; 3-truncated"
     CDS             complement(625..1458)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWSSGGGPVPNSPYSESYYARSLAVVLQ
                     RRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGARLQGST
                     GQGVTPSMHGFNGGHSSPLGDLQQLSCIQLQRWSSEFTSHSQQSLQSTSTGPLLAHSND
                     GFVGSLRQGSDHILVLDSHCSESSWVLAQSHDVSVSEAVTNQGFVVLSDGRAHLGLDLL
                     LLLLGVAQSAQVNHRDSVDAHFTLLLGSHLVVVVLVVDVVVLVVAVVVLVIHFDF"
     CDS             637..1152
                     /codon_start=1
                     /product="TAL DNA-binding domain"
                     /label=TAL DNA-binding domain
                     /note="3-truncated; contains TAL N-terminus"
                     /protein_id="AHX83888.1"
                     /translation="MDYKDHDGDYKDHDIDYKDDDDKMAPKKKRKVGIHGVPMVDLRTL
                     GYSQQQQEKIKPKVRSTVAQHHEALVGHGFTHAHIVALSQHPAALGTVAVKYQDMIAAL
                     PEATHEAIVGVGKQWSGARALEALLTVAGELRGPPLQLDTGQLLKIAKRGGVTAVEAVH
                     AWRNALTGA"
     misc_feature    complement(1159..1164)
                     /label=BsmBI restriction site
                     /note="BsmBI restriction site"
     CDS             complement(1165..1458)
                     /codon_start=1
                     /product="LacZ-like protein"
                     /label=LacZ-like protein
                     /protein_id="AHX83889.1"
                     /translation="MTMITPSAQLTLTKGNKSWSSGGGPVPNSPYSESYYARSLAVVLQ
                     RRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSGA"
     primer_bind     1326..1342
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1352..1370
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1415..1433)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1454..1470)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1478..1494
                     /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(1502..1532)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1547..1568
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    1603..1608
                     /label=BsmBI restriction site
                     /note="BsmBI restriction site"
     CDS             1610..2350
                     /codon_start=1
                     /product="TAL/Fok I nuclease"
                     /label=TAL/Fok I nuclease
                     /note="5-truncated; contains TAL C-terminus"
                     /protein_id="AHX83890.1"
                     /translation="NDHLVALACLGGRPALDAVKKGLPHAPALIKRTNRRIPERTSHRV
                     AGSQLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKVMEFFMKVYGYR
                     GKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQRYVEENQTRNKH
                     INPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVLSVEELLIGGEM
                     IKAGTLTLEEVRRKFNNGEINFRS"
     CDS             1754..2341
                     /codon_start=1
                     /product="nonspecific DNA cleavage domain of the FokI 
                     endonuclease (Li et al., 1992)"
                     /label=FokI cleavage domain
                     /note="must dimerize to be catalytically active"
                     /translation="QLVKSELEEKKSELRHKLKYVPHEYIELIEIARNSTQDRILEMKV
                     MEFFMKVYGYRGKHLGGSRKPDGAIYTVGSPIDYGVIVDTKAYSGGYNLPIGQADEMQR
                     YVEENQTRNKHINPNEWWKVYPSSVTEFKFLFVSGHFKGNYKAQLTRLNHITNCNGAVL
                     SVEELLIGGEMIKAGTLTLEEVRRKFNNGEINF"
     polyA_signal    2401..2482
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    2590..532
                     /label=pBluescript SK+
                     /note="pBluescript SK+"
     rep_origin      complement(2826..3414)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3585..4445)
                     /codon_start=1
                     /gene="ampR"
                     /product="AmpR"
                     /label=ampR
                     /protein_id="AHX83891.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(3585..4445)
                     /gene="ampR"
                     /label=ampR
     promoter        complement(4446..4550)
                     /gene="bla"
                     /label=AmpR promoter