LOCUS       Exported                8772 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pAP151, complete sequence.
ACCESSION   MF467525
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8772)
  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  JOURNAL   Unpublished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 8772)
  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 8772)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8772
                     /organism="Cloning vector pAP151"
                     /mol_type="other DNA"
                     /label=derived from Mycobacterium smegmatis str. MC2 155
                     /note="derived from Mycobacterium smegmatis str. MC2 155"
                     /db_xref="taxon:2204079"
     terminator      37..83
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     359..375
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    395..415
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     CDS             complement(451..1074)
                     /codon_start=1
                     /product="TetR"
                     /label=TetR
                     /note="Tet repressor protein"
                     /protein_id="AWN08190.1"
                     /translation="MSRLDKSKVINSALALGNEVGIEGVTTRKLAQKLGVAQKTLYWHV
                     KNKRALLDALAVEILARHHDYSLPAAGESWQSFLRNNAMSFRRALLRYRDGAKVHLGTR
                     PDEKQYDTVETQLRFMTENGFSLENALYALSAVGHFTLGCVLEDQEHTAAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     regulatory      complement(1107..1600)
                     /regulatory_class="promoter"
     protein_bind    1600..1624
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1605..1798
                     /regulatory_class="promoter"
     protein_bind    complement(1799..1823)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1838..4429
                     /codon_start=1
                     /product="ClpC1-DAS"
                     /label=ClpC1-DAS
                     /note="ClpC1 ATPase with DAS tag at C terminus"
                     /protein_id="AWN08191.1"
                     /translation="MFERFTDRARRVVVLAQEEARMLNHNYIGTEHILLGLIHEGEGVA
                     AKSLESLGISLEGVRSQVEEIIGQGQQAPSGHIPFTPRAKKVLELSLREALQLGHNYIG
                     TEHILLGLIREGEGVAAQVLVKLGAELTRVRQQVIQLLSGYQGKEAAEAGTGGRGGESG
                     SPSTSLVLDQFGRNLTAAAMEGKLDPVIGREKEIERVMQVLSRRTKNNPVLIGEPGVGK
                     TAVVEGLAQAIVHGEVPETLKDKQLYTLDLGSLVAGSRYRGDFEERLKKVLKEINTRGD
                     IILFIDELHTLVGAGAAEGAIDAASILKPKLARGELQTIGATTLDEYRKYIEKDAALER
                     RFQPVQVGEPTVEHTIEILKGLRDRYEAHHRVSITDAAMVAAATLADRYINDRFLPDKA
                     IDLIDEAGARMRIRRMTAPPDLREFDEKIAEARREKESAIDAQDFEKAASLRDREKTLV
                     AQRAEREKQWRSGDLDVVAEVDDEQIAEVLGNWTGIPVFKLTEAETTRLLRMEEELHKR
                     IIGQEDAVKAVSKAIRRTRAGLKDPKRPSGSFIFAGPSGVGKTELSKALANFLFGDDDA
                     LIQIDMGEFHDRFTASRLFGAPPGYVGYEEGGQLTEKVRRKPFSVVLFDEIEKAHQEIY
                     NSLLQVLEDGRLTDGQGRTVDFKNTVLIFTSNLGTSDISKPVGLGFSKGGGENDYERMK
                     QKVNDELKKHFRPEFLNRIDDIIVFHQLTREEIIRMVDLMISRVAGQLKSKDMALVLTD
                     AAKALLAKRGFDPVLGARPLRRTIQREIEDQLSEKILFEEVGPGQVVTVDVDNWDGEGP
                     GEDAVFTFTGTRKPPAEPDLAKAGAHSAGGPEPAARAANDENYSENYADAS"
     polyA_signal    4801..4935
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(5131..5719)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6051..6866)
                     /codon_start=1
                     /product="Aph"
                     /label=Aph
                     /note="kanamycin resistance protein"
                     /protein_id="AWN08192.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPHAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHNLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(7147..8262)
                     /codon_start=1
                     /product="L5 integrase"
                     /label=L5 integrase
                     /note="integrates plasmid into the L5 chromosomal 
                     integration site"
                     /protein_id="AWN08193.1"
                     /translation="MARRGWGSLKTQRSGRIQASYVNPQDGVRYYALQTYDNKMDAEAW
                     LAGEKRLIEMETWTPPQDRAKKAAASAITLEEYTRKWLVERDLADGTRDLYSGHAERRI
                     YPVLGEVAVTEMTPALVRAWWAGMGRKHPTARRHAYNVLRAVMNTAVEDKLIAENPCRI
                     EQKAADERDVEALTPEELDIVAAEIFEHYRIAAYILAWTSLRFGELIELRRKDIVDDGM
                     TMKLRVRRGASRVGNKIVVGNAKTVRSKRPVTVPPHVAEMIRAHMKDRTKMNKGPEAFL
                     VTTTQGNRLSKSAFTKSLKRGYAKIGRPELRIHDLRAVGATFAAQAGATTKELMARLGH
                     TTPRMAMKYQMASEARDEAIAEAMSKLAKTS"