LOCUS       Exported                3903 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector patt-shyg-oriT, complete sequence.
ACCESSION   KF488575
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3903)
  AUTHORS   Myronovskyi M, Rosenkranzer B, Luzhetskyy A.
  TITLE     Iterative marker excision system
  JOURNAL   Appl. Microbiol. Biotechnol. 98 (10), 4557-4570 (2014)
  PUBMED    24473925
REFERENCE   2  (bases 1 to 3903)
  AUTHORS   Myronovskyi M, Rosenkraenzer B, Luzhetskyy A.
  TITLE     Direct Submission
  JOURNAL   Submitted (31-JUL-2013) AMEG, Helmholtz Institute for Pharmaceutical
            Research Saarland, Campus, Geb. C2.3 Universitaet des Saarlandes, 
            Saarbruecken, Saarland 66123, Germany
REFERENCE   3  (bases 1 to 3903)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..3903
                     /organism="Cloning vector patt-shyg-oriT"
                     /mol_type="other DNA"
                     /db_xref="taxon:1406238"
     primer_bind     293..309
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        320..338
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     383..402
     misc_recomb     complement(409..459)
                     /label=B-CC
                     /note="B-CC"
     RBS             566..574
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3 end of the 16S rRNA (Chen et al., 1994)"
     CDS             580..1578
                     /codon_start=1
                     /gene="hyg"
                     /product="hygromycin phosphotransferase"
                     /label=hyg
                     /protein_id="AGX01515.1"
                     /translation="MTQESLLLLDRIDSDDSYASLRNDQEFWEPLARRALEELGLPVPP
                     VLRVPGESTNPVLVGEPDPVIKLFGEHWCGPESLASESEAYAVLADAPVPVPRLLGRGE
                     LRPGTGAWPWPYLVMSRMTGTTWRSAMDGTTDRNALLALARELGRVLGRLHRVPLTGNT
                     VLTPHSEVFPELLRERRAATVEDHRGWGYLSPRLLDRLEDWLPDVDTLLAGREPRFVHG
                     DLHGTNIFVDLAATEVTGIVDFTDVYAGDSRYSLVQLHLNAFRGDREILAALLDGAQWK
                     RTEDFARELLAFTFLHDFEVFEETPLDLSGFTDPEELAQFLWGPPDTAPGA"
     gene            580..1578
                     /gene="hyg"
                     /label=hyg
     misc_feature    1588..1829
                     /label=oriT
                     /note="oriT"
     misc_recomb     1842..1891
                     /label=P-GG
                     /note="P-GG"
     primer_bind     complement(1898..1917)
     rep_origin      2100..2767
     rep_origin      complement(2156..2744)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2915..3775)
                     /codon_start=1
                     /gene="ampR"
                     /product="beta-lactamase"
                     /label=ampR
                     /protein_id="AGX01516.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(2915..3775)
                     /gene="ampR"
                     /label=ampR
     promoter        complement(3776..3880)
                     /gene="bla"
                     /label=AmpR promoter