LOCUS       Exported                5722 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pAP258, complete sequence.
ACCESSION   MF467527
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5722)
  AUTHORS   Park AS, Rubin EJ.
  TITLE     Clp-mediated Regulation of WhiB1 is Required for Cell Division in 
            Mycobacteria
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5722)
  AUTHORS   Park AS, Rubin EJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-JUL-2017) Department of Immunology and Infectious 
            Diseases, Harvard School of Public Health, 665 Huntington Ave, 
            Boston, MA 02115, USA
REFERENCE   3  (bases 1 to 5722)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5722
                     /organism="Cloning vector pAP258"
                     /mol_type="other DNA"
                     /label=derived from Mycobacterium smegmatis str. MC2 155
                     /note="derived from Mycobacterium smegmatis str. MC2 155"
                     /db_xref="taxon:2204081"
     terminator      41..68
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     regulatory      93..398
                     /regulatory_class="promoter"
     protein_bind    268..285
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     regulatory      414..419
                     /regulatory_class="ribosome_binding_site"
     CDS             427..681
                     /codon_start=1
                     /product="WhiB1 (G61E)"
                     /label=WhiB1
                     /note="transcriptional regulator with mutation G61E"
                     /protein_id="AWN08197.1"
                     /translation="MDWRHKAVCRDEDPELFFPVGNSGPALAQIADAKLVCNRCPVTTE
                     CLSWALESGQDAGVWEGMSEDERRALKRRNARTKARTGV"
     regulatory      complement(737..768)
                     /regulatory_class="terminator"
     CDS             complement(799..1422)
                     /codon_start=1
                     /product="TetR"
                     /label=TetR
                     /note="tetracycline repressor class B"
                     /protein_id="AWN08198.1"
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     regulatory      complement(1449..1765)
                     /regulatory_class="promoter"
     regulatory      1784..1811
                     /regulatory_class="terminator"
     terminator      1784..1811
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      complement(1993..2581)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2913..3728)
                     /codon_start=1
                     /gene="aph(3)-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPHAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHNLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4009..5124)
                     /codon_start=1
                     /product="L5 integrase"
                     /label=L5 integrase
                     /protein_id="AWN08199.1"
                     /translation="MARRGWGSLKTQRSGRIQASYVNPQDGVRYYALQTYDNKMDAEAW
                     LAGEKRLIEMETWTPPQDRAKKAAASAITLEEYTRKWLVERDLADGTRDLYSGHAERRI
                     YPVLGEVAVTEMTPALVRAWWAGMGRKHPTARRHAYNVLRAVMNTAVEDKLIAENPCRI
                     EQKAADERDVEALTPEELDIVAAEIFEHYRIAAYILAWTSLRFGELIELRRKDIVDDGM
                     TMKLRVRRGASRVGNKIVVGNAKTVRSKRPVTVPPHVAEMIRAHMKDRTKMNKGPEAFL
                     VTTTQGNRLSKSAFTKSLKRGYAKIGRPELRIHDLRAVGATFAAQAGATTKELMARLGH
                     TTPRMAMKYQMASEARDEAIAEAMSKLAKTS"
     terminator      5671..5717
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      complement(5671..5714)
                     /regulatory_class="terminator"