LOCUS       Exported                3810 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pST76A-SceP, complete sequence.
ACCESSION   AF170482
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3810)
  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 3810)
  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 3810)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3810
                     /organism="Cloning vector pST76A-SceP"
                     /lab_host="Escherichia coli"
                     /mol_type="genomic DNA"
                     /note="stimulates gene replacement; low copy number"
                     /db_xref="taxon:102131"
     promoter        57..112
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     regulatory      58..63
                     /regulatory_class="minus_35_signal"
                     /note="tetA promoter from transposon Tn10"
     protein_bind    63..81
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     misc_binding    73..81
                     /label=TetR binding site
                     /bound_moiety="TetR"
                     /note="site A"
     regulatory      82..88
                     /regulatory_class="minus_10_signal"
                     /note="tetA promoter from transposon Tn10"
     protein_bind    93..111
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_binding    103..111
                     /label=TetR binding site
                     /bound_moiety="TetR"
                     /note="site B"
     CDS             120..821
                     /codon_start=1
                     /gene="I-SceI"
                     /product="I-SceI meganuclease"
                     /label=I-SceI
                     /protein_id="AAD51640.1"
                     /translation="MHQKNQVMNLGPNSKLLKEYKSQLIELNIEQFEAGIGLILGDAYI
                     RSRDEGKTYCMQFEWKNKAYMDHVCLLYDQWVLSPPHKKQRVNHLGNLVITWGAQTFKH
                     QAFNKLANLFIVNNKKTIPNNLVENYLTPMSLAYWFMDDGGKWDYNKNSTNKSIVLNTQ
                     SFTFEEVEYLVKGLRNKFQLNCYVKINKNKPIIYIDSMSYLIFYNLIKPYLIPQMMYKL
                     PNTISSETFLK"
     gene            120..821
                     /gene="I-SceI"
                     /label=I-SceI
     CDS             complement(976..1926)
                     /codon_start=1
                     /gene="rep ts"
                     /product="Rep ts replication protein"
                     /label=rep ts
                     /note="derived from pHS1"
                     /protein_id="AAD51641.1"
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     gene            complement(976..1926)
                     /gene="rep ts"
                     /label=rep ts
     rep_origin      1927..2306
                     /label=derived from pHS1
                     /note="derived from pHS1"
     rep_origin      1974..2196
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             complement(2818..3678)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="AAD51642.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3679..3783)
                     /gene="bla"
                     /label=AmpR promoter