LOCUS       Exported                3474 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pUC19RP12, complete sequence.
ACCESSION   AF170481
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3474)
  AUTHORS   Posfai G, Kolisnychenko V, Bereczki Z, Blattner FR.
  TITLE     Markerless gene replacement in Escherichia coli stimulated by a 
            double-strand break in the chromosome
  JOURNAL   Nucleic Acids Res. 27 (22), 4409-4415 (1999)
  PUBMED    10536150
REFERENCE   2  (bases 1 to 3474)
  AUTHORS   Posfai G, Kolisnychenko V, Bereczki Z, Blattner FR.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-JUL-1999) Institute of Biochemistry, Biol. Res. Center
            Hung. Acad.Sci., Temesvari krt.,62, Szeged H-6726, Hungary
REFERENCE   3  (bases 1 to 3474)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3474
                     /organism="Cloning vector pUC19RP12"
                     /lab_host="Escherichia coli"
                     /mol_type="genomic DNA"
                     /note="stimulates gene replacement; high copy number"
                     /db_xref="taxon:102133"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        450..505
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     regulatory      451..456
                     /regulatory_class="minus_35_signal"
                     /note="tetA promoter from transposon Tn10"
     protein_bind    456..474
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     misc_binding    466..474
                     /label=TetR binding site
                     /bound_moiety="TetR"
                     /note="site A"
     regulatory      475..481
                     /regulatory_class="minus_10_signal"
                     /note="tetA promoter from transposon Tn10"
     protein_bind    486..504
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_binding    496..502
                     /label=TetR binding site
                     /bound_moiety="TetR"
                     /note="site B"
     CDS             513..1214
                     /codon_start=1
                     /gene="I-SceI"
                     /product="I-SceI meganuclease"
                     /label=I-SceI
                     /protein_id="AAD51638.1"
                     /translation="MHQKNQVMNLGPNSKLLKEYKSQLIELNIEQFEAGIGLILGDAYI
                     RSRDEGKTYCMQFEWKNKAYMDHVCLLYDQWVLSPPHKKQRVNHLGNLVITWGAQTFKH
                     QAFNKLANLFIVNNKKTIPNNLVENYLTPMSLAYWFMDDGGKWDYNKNSTNKSIVLNTQ
                     SFTFEEVEYLVKGLRNKFQLNCYVKINKNKPIIYIDSMSYLIFYNLIKPYLIPQMMYKL
                     PNTISSETFLK"
     gene            513..1214
                     /gene="I-SceI"
                     /label=I-SceI
     primer_bind     complement(1253..1269)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1277..1293
                     /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(1301..1331)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1346..1367
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      1652
                     /label=ColE1
                     /note="ColE1"
     rep_origin      complement(1655..2243)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2414..3274)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="AAD51639.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3275..3379)
                     /gene="bla"
                     /label=AmpR promoter