LOCUS       Exported                7366 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Integrating expression vector pALE2, complete sequence.
ACCESSION   DQ086480
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7366)
  AUTHORS   Fortune SM, Jaeger A, Sarracino DA, Chase MR, Sassetti CM, Sherman 
            DR, Bloom BR, Rubin EJ.
  TITLE     Mutually dependent secretion of proteins required for mycobacterial 
            virulence
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 102 (30), 10676-10681 (2005)
  PUBMED    16030141
REFERENCE   2  (bases 1 to 7366)
  AUTHORS   Fortune SM, Jaeger A, Sherman DR, Rubin EJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-JUN-2005) IID, Harvard School of Public Health, 200 
            Longwood Ave., Boston, MA 02115, USA
REFERENCE   3  (bases 1 to 7366)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7366
                     /organism="Integrating expression vector pALE2"
                     /mol_type="other DNA"
                     /label=derived from Mycobacterium tuberculosis
                     /note="derived from Mycobacterium tuberculosis"
                     /db_xref="taxon:333346"
     terminator      52..138
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      230..257
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        303..333
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    339..358
                     /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."
     CDS             387..1625
                     /codon_start=1
                     /gene="Rv3616c"
                     /product="Rv3616c-hismyc"
                     /label=Rv3616c
                     /note="C-terminus tagged with his and myc from synthetic 
                     MOP promoter"
                     /protein_id="AAY86710.1"
                     /translation="MSRAFIIDPTISAIDGLYDLLGIGIPNQGGILYSSLEYFEKALEE
                     LAAAFPGDGWLGSAADKYAGKNRNHVNFFQELADLDRQLISLIHDQANAVQTTRDILEG
                     AKKGLEFVRPVAVDLTYIPVVGHALSAAFQAPFCAGAMAVVGGALAYLVVKTLINATQL
                     LKLLAKLAELVAAAIADIISDVADIIKGTLGEVWEFITNALNGLKELWDKLTGWVTGLF
                     SRGWSNLESFFAGVPGLTGATSGLSQVTGLFGAAGLSASSGLAHADSLASSASLPALAG
                     IGGGSGFGGLPSLAQVHAASTRQALRPRADGPVGAAAEQVGGQSQLVSAQGSQGMGGPV
                     GMGGMHPSSGASKGTTTKKYSEGAAAGTEDAERAPVEADAGGGQKVLVRNVVTSHHHHH
                     HMAEQKLISEEDL"
     gene            387..1625
                     /gene="Rv3616c"
                     /label=Rv3616c
     CDS             1569..1586
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1593..1622
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      1666..1713
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      complement(1917..2505)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2824..3483)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"