LOCUS       Exported                5460 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pAP264, complete sequence.
ACCESSION   MF510148
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5460)
  AUTHORS   Park AS, Rubin EJ, Minch KJ, Sherman DR.
  TITLE     Clp-mediated Regulation of WhiB1 is Required for Cell Division in 
            Mycobacteria
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5460)
  AUTHORS   Park AS, Rubin EJ, Minch KJ, Sherman DR.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-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 5460)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5460
                     /organism="Cloning vector pAP264"
                     /mol_type="other DNA"
                     /db_xref="taxon:2212997"
     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"
     regulatory      complement(475..506)
                     /regulatory_class="terminator"
     CDS             complement(537..1160)
                     /codon_start=1
                     /product="TetR"
                     /label=TetR
                     /note="Tet repressor class B"
                     /protein_id="AWT04829.1"
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     regulatory      complement(1187..1503)
                     /regulatory_class="promoter"
     regulatory      1522..1549
                     /regulatory_class="terminator"
     terminator      1522..1549
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      1716..2335
     rep_origin      complement(1731..2319)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2651..3466)
                     /codon_start=1
                     /product="Aph"
                     /label=Aph
                     /note="kanamycin resistance protein"
                     /protein_id="AWT04830.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPHAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHNLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3747..4745)
                     /codon_start=1
                     /product="L5 integrase"
                     /label=L5 integrase
                     /protein_id="AWT04831.1"
                     /translation="MDAEAWLAGEKRLIEMETWTPPQDRAKKAAASAITLEEYTRKWLV
                     ERDLADGTRDLYSGHAERRIYPVLGEVAVTEMTPALVRAWWAGMGRKHPTARRHAYNVL
                     RAVMNTAVEDKLIAENPCRIEQKAADERDVEALTPEELDIVAAEIFEHYRIAAYILAWT
                     SLRFGELIELRRKDIVDDGMTMKLRVRRGASRVGNKIVVGNAKTVRSKRPVTVPPHVAE
                     MIRAHMKDRTKMNKGPEAFLVTTTQGNRLSKSAFTKSLKRGYAKIGRPELRIHDLRAVG
                     ATFAAQAGATTKELMARLGHTTPRMAMKYQMASEARDEAIAEAMSKLAKTS"
     terminator      5409..5455
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      5409..5452
                     /regulatory_class="terminator"