LOCUS       Exported                4608 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  PCR template vector pTF104 DNA, complete sequence.
ACCESSION   AB303314
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4608)
  AUTHORS   Fukuda T, Ohya Y, Ohta K.
  TITLE     Conditional genomic rearrangement by designed meiotic recombination 
            using VDE (PI-SceI) in yeast
  JOURNAL   Mol. Genet. Genomics 278 (4), 467-478 (2007)
  PUBMED    17587055
REFERENCE   2  (bases 1 to 4608)
  AUTHORS   Fukuda T, Ohta K.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-MAY-2007) Tomoyuki Fukuda, RIKEN; 2-1 Hirosawa, Wako, 
            Saitama 351-0198, Japan (E-mail:tfukuda@riken.jp, 
            Tel:81-48-467-9277(ex.5552), Fax:81-48-462-4691)
REFERENCE   3  (bases 1 to 4608)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4608
                     /organism="PCR template vector pTF104"
                     /mol_type="other DNA"
                     /db_xref="taxon:442843"
     misc_feature    1..103
                     /note="sequence containing a part of the recognition 
                     sequence of VDE"
     gene            complement(167..1523)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     terminator      167..364
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(370..1179)
                     /codon_start=1
                     /gene="KanR"
                     /product="aminoglycoside 3-phosphotransferase"
                     /label=KanR
                     /note="kanamycin and geneticin G418 resistance protein"
                     /protein_id="BAF62427.1"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            complement(370..1179)
                     /gene="KanR"
                     /label=KanR
     promoter        complement(1180..1523)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     misc_feature    1607..1707
                     /note="sequence containing a part of the recognition 
                     sequence of VDE"
     promoter        complement(1737..1755)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1762..1778)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      1920..2375
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2401..2505
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2506..3366
                     /codon_start=1
                     /gene="AmpR"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="E. coli ampicillin resistance gene"
                     /protein_id="BAF62428.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            2506..3366
                     /gene="AmpR"
                     /label=AmpR
     rep_origin      3537..4125
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    4413..4434
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4449..4479
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4487..4503
                     /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     4511..4527
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4548..4566
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"