LOCUS       Exported                6945 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Integrating expression vector pJEB403+drrA, complete sequence.
ACCESSION   FJ009257
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6945)
  AUTHORS   Murry JP, Pandey AK, Sassetti CM, Rubin EJ.
  TITLE     Phthiocerol Dimycocerosate Transport Is Required for Resisting 
            Interferon-gamma-Independent Immunity
  JOURNAL   J. Infect. Dis. 200 (5), 774-782 (2009)
  PUBMED    19622047
REFERENCE   2  (bases 1 to 6945)
  AUTHORS   Murry JP, Rubin EJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-AUG-2008) Microbiology and Molecular Genetics, Harvard
            Medical School, 200 Longwood Ave., Boston, MA 02115, USA
REFERENCE   3  (bases 1 to 6945)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6945
                     /organism="Integrating expression vector pJEB403+drrA"
                     /mol_type="other DNA"
                     /db_xref="taxon:586732"
     misc_feature    218..477
                     /label=attP
                     /note="attP"
     CDS             629..1627
                     /codon_start=1
                     /gene="int"
                     /product="integrase"
                     /label=int
                     /note="L5 mycobacterial phage integrase"
                     /protein_id="ACL30946.1"
                     /translation="MDAEAWLAGEKRLIEMETWTPPQDRAKKAAASAITLEEYTRKWLV
                     ERDLADGTRDLYSGHAERRIYPVLGEVAVTEMTPALVRAWWAGMGRKHPTARRHAYNVL
                     RAVMNTAVEDKLIAENPCRIEQKAADERDVEALTPEELDIVAAEIFEHYRIAAYILAWT
                     SLRFGELIELRRKDIVHDGMTMKLRVRRGASRVGNKIVVGNAKTVRSKRPVTVPPHVAE
                     MIRAHMKDRTKMNKGPEAFLVTTTQGNRLSKSAFTKSLKRGYAKIGRPELRIHDLRAVG
                     ATFAAQAGATTKELMARLGHTTPRMAMKYQMASEARDEAIAEAMSKLAKTS"
     gene            629..1627
                     /gene="int"
                     /label=int
     primer_bind     2711..2727
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2736..2769
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(2807..3466)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /protein_id="ACL30947.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(2807..3466)
                     /gene="cat"
                     /label=cat
     promoter        complement(3467..3569)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             complement(3567..4565)
                     /codon_start=1
                     /gene="aph"
                     /product="Aph"
                     /label=aph
                     /note="hygromycin resistance"
                     /protein_id="ACL30948.1"
                     /translation="MTQESLLLLDRIDSDDSYASLRNDQEFWEPLARRALEELGLPVPP
                     VLRVPGESTNPVLVGEPDPVIKLFGEHWCGPESLASESEAYAVLADAPVPVPRLLGRGE
                     LRPGTGAWPWPYLVMSRMTGTTWRSAMDGTTDRNALLALARELGRVLGRLHRVPLTGNT
                     VLTPHSEVFPELLRERRAATVEDHRGWGYLSPRLLDRLEDWLPDVDTLLAGREPRFVHG
                     DLHGTNIFVDLAATEVTGIVDFTDVYAGDSRYSLVQLHLNAFRGDREILAALLDGAQWK
                     RTEDFARELLAFTFLHDFEVFEETPLDLSGFTDPEELAQFLWGPPDTAPGA"
     gene            complement(3567..4565)
                     /gene="aph"
                     /label=aph
     protein_bind    4686..4719
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     rep_origin      4725..5518
                     /label=pUC origin of replication
                     /note="pUC origin of replication"
     rep_origin      4775..5363
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5553..6548)
                     /codon_start=1
                     /gene="drrA"
                     /product="DrrA"
                     /label=drrA
                     /note="ABC transporter subunit involved in phthiocerol 
                     dimycocerosate transport"
                     /protein_id="ACL30949.1"
                     /translation="MRNDDMAVVVNGVRKTYGKGKIVALDDVSFKVRRGEVIGLLGPNG
                     AGKTTMVDILSTLTRPDAGSAIIAGYDVVSEPAGVRRSIMVTGQQVAVDDALSGEQNLV
                     LFGRLWGLSKSAARKRAAELLEQFSLVHAGKRRVGTYSGGMRRRIDIACGLVVQPQVAF
                     LDEPTTGLDPRSRQAIWDLVASFKKLGIATLLTTQYLEEADALSDRIILIDHGIIIAEG
                     TANELKHRAGDTFCEIVPRDLKDLDAIVAALGSLLPEHHRAMLTPDSDRITMPAPDGIR
                     MLVEAARRIDEARIELADIALRRPSLDHVFLAMTTDPTESLTHLVSGSAR"
     gene            complement(5553..6548)
                     /gene="drrA"
                     /label=drrA
     regulatory      complement(6576..6652)
                     /regulatory_class="promoter"
                     /note="mop"
     protein_bind    6588..6607
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     promoter        complement(6613..6643)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     terminator      6689..6716
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      6808..6894
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"