LOCUS       Exported                5315 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Gene deletion vector pYS2, complete sequence.
ACCESSION   KF582041
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5315)
  AUTHORS   Shenkerman Y, Elharar Y, Vishkautzan M, Gur E.
  TITLE     Efficient and simple generation of unmarked gene deletions in 
            Mycobacterium smegmatis
  JOURNAL   Gene 533 (1), 374-378 (2014)
  PUBMED    24100088
REFERENCE   2  (bases 1 to 5315)
  AUTHORS   Shenkerman Y, Elharar Y, Vishkautzan M, Gur E.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-AUG-2013) Life Sciences, Ben-Gurion University of the 
            Negev, Rager Blvd, Beer-Sheva 84105, Israel
REFERENCE   3  (bases 1 to 5315)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5315
                     /organism="Gene deletion vector pYS2"
                     /lab_host="Mycobacterium smegmatis"
                     /mol_type="other DNA"
                     /db_xref="taxon:1408816"
     protein_bind    106..127
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        142..172
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(616..649)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_recomb     complement(618..649)
                     /label=loxP site for cre recombinase
                     /note="loxP site for cre recombinase"
     CDS             1008..1724
                     /codon_start=1
                     /gene="mycgfp2+"
                     /product="GFP"
                     /label=mycgfp2+
                     /protein_id="AGY30915.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKTRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     gene            1008..1724
                     /gene="mycgfp2+"
                     /label=mycgfp2+
     CDS             1836..2834
                     /codon_start=1
                     /gene="hyg"
                     /product="Hyg"
                     /label=hyg
                     /note="hygromycin resistance"
                     /protein_id="AGY30914.1"
                     /translation="MTQESLLLLDRIDSDDSYASLRNDQEFWEPLARRALEELGLPVPP
                     VLRVPGESTNPVLVGEPDPVIKLFGEHWCGPESLASESEAYAVLADAPVPVPRLLGRGE
                     LRPGTGAWPWPYLVMSRMTGTTWRSAMDGTTDRNALLALARELGRVLGRLHRVPLTGNT
                     VLTPHSEVFPELLRERRAATVEDHRGWGYLSPRLLDRLEDWLPDVDTLLAGREPRFVHG
                     DLHGTNIFVDLAATEVTGIVDFTDVYAGDSRYSLVQLHLNAFRGDREILAALLDGAQWK
                     RTEDFARELLAFTFLHDFEVFEETPLDLSGFTDPEELAQFLWGPPDTAPGA"
     gene            1836..2834
                     /gene="hyg"
                     /label=hyg
     misc_recomb     3059..3092
                     /label=loxP site for cre recombinase
                     /note="loxP site for cre recombinase"
     protein_bind    complement(3059..3092)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        3409..3513
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3514..4374
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             3712..4374
                     /codon_start=1
                     /gene="bla"
                     /product="bla"
                     /label=bla
                     /note="ampicillin resistance"
                     /protein_id="AGY30913.1"
                     /translation="MSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHL
                     TDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEA
                     IPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFI
                     ADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     gene            3712..4374
                     /gene="bla"
                     /label=bla
     rep_origin      4469..5151
                     /label=ColE1 origin
                     /note="ColE1 origin"
     rep_origin      4545..5133
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"