LOCUS       Exported                7692 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Escherichia-Pseudomonas shuttle vector pCon2(3), complete sequence.
ACCESSION   JF276396
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7692)
  AUTHORS   Raja Abd Rahman RNZ., Wong CF, Salleh AB, Basri M.
  TITLE     Functions of Plac and PT7(A1/04/03) in overexpression of elastase 
            strain K by E. coli and Pseudomonas sp. harbouring pCon2(3)##
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7692)
  AUTHORS   Raja Abd Rahman RNZ., Wong CF, Salleh AB, Basri M.
  TITLE     Direct Submission
  JOURNAL   Submitted (31-JAN-2011) Department of Microbiology, Universiti Putra
            Malaysia, Faculty of Biotechnology and Biomolecular Sciences, UPM 
            Serdang, Selangor 43400, Malaysia
REFERENCE   3  (bases 1 to 7692)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7692
                     /organism="Escherichia-Pseudomonas shuttle vector
                     pCon2(3)"
                     /mol_type="other DNA"
                     /db_xref="taxon:999508"
     CDS             complement(314..1810)
                     /codon_start=1
                     /product="organic solvent tolerant elastase"
                     /label=organic solvent tolerant elastase
                     /note="prepro-elastase; derived from the sequence of INSD 
                     accession EU021222"
                     /protein_id="AEA29842.1"
                     /translation="MKKVSTLDLLFVAIMGVSPAALAADLIDVSKLPSKAAQGAPGPVT
                     LQAAVGAGGADELKAIRSTTLPNGKQVTRYEQFHNGVRVVGEAITEVKGPGKSVAAQRS
                     GHFVANIAADLPGSTTAAVSAEQVLAQAKSLKAQGRKTENDKVELVIRLGESNIAQLVY
                     NVSYLIPGEGLSRPHFVIDAKTGEVLDQWEGLAHAEAGGPGGNQKIGKYTYGSDYGPLI
                     VNDRCEMDDGNVITVDMNGSTDDSKTTPFRFACPTNTYKQVNGAYSPLNDAHFFGGVVF
                     KLYRDWFGTSPLTHKLYMKVHYGRSVENAYWGGTAMLFGDGATMFYPLVSLDVAAHEVS
                     HGFTEQNSGLIYRGQSGGMNEAFSDMAGEAAEFYMRGKNDFLIGYDIKKGSGALRYMDQ
                     PSRDGRSIDNASQYYNGIDVHHSSGVYNRAFYLLANSPGWDTRKAFEVFVDANRYYWTA
                     TSNYNSGACGVIRSAQNRNYSAADVTRAFSTVGVTCPSAL"
     misc_feature    complement(1856..1884)
                     /label=operator O3 of PT7(A1/O4/O3)
                     /note="operator O3 of PT7(A1/O4/O3)"
     protein_bind    1866..1882
                     /label=lac operator
                     /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)."
     misc_feature    complement(1892..1897)
                     /label=promoter -10 of (PT7(A1/04/03)
                     /note="promoter -10 of (PT7(A1/04/03)"
     misc_feature    complement(1898..1914)
                     /label=operator O4 of PT7(A1/O4/O3)
                     /note="operator O4 of PT7(A1/O4/O3)"
     protein_bind    1899..1915
                     /label=lac operator
                     /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)."
     misc_feature    complement(1915..1920)
                     /label=promoter -33 of PT7(A1/O4/O3)
                     /note="promoter -33 of PT7(A1/O4/O3)"
     misc_feature    1998..3879
                     /label=SF
                     /note="SF"
     rep_origin      2649..3000
                     /label=pRO1600 oriV
                     /note="broad-host-range origin of replication from 
                     Pseudomonas aeruginosa plasmid pRO1600; requires the 
                     pRO1600 Rep protein for replication (West et al., 1994)"
     CDS             3014..3847
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pRO1600 from Pseudomonas aeruginosa "
                     /label=pRO1600 Rep
                     /translation="MASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
     primer_bind     4008..4024
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(4072..5568)
                     /codon_start=1
                     /product="organic solvent tolerant elastase"
                     /label=organic solvent tolerant elastase
                     /note="prepro-elastase; derived from the sequence of INSD 
                     accession EU021222"
                     /protein_id="AEA29843.1"
                     /translation="MKKVSTLDLLFVAIMGVSPAAFAADLIDVSKLPSKAAQGAPGPVT
                     LQAAVGAGGADELKAIRSTTLPNGKQVTRYEQFHNGVRVVGEAITEVKGPGKSVAAQRS
                     GHFVANIAADLPGSTTAAVSAEQVLAQAKSLKAQGRKTENDKVELVIRLGENNIAQLVY
                     NVSYLIPGEGLSRPHFVIDAKTGEVLDQWEGLAHAEAGGPGGNQKIGKYTYGSDYGPLI
                     VNDRCEMDDGNVITVDMNGSTDDSKTTPFRFACPTNTYKQVNGAYSPLNDAHFFGGVVF
                     KLYRDWFGTSPLTHKLYMKVHYGRSVENAYWDGTAMLFGDGATMFYPLVSLDVAAHEVS
                     HGFTEQNSGLIYRGQSGGMNEAFSDMAGEAAEFYMRGKNDFLIGYDIKKGSGALRYMDQ
                     PSRDGRSIDNASQYYNGIDVHHSSGVYNRAFYLLANSPGWDTRKAFEVFVDANRYYWTA
                     TSNYNSGACGVIRSAQNRNYSAADVTRAFSTVGVTCPSAL"
     primer_bind     complement(5588..5604)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5612..5628
                     /label=lac operator
                     /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)."
     misc_feature    complement(5628..5632)
                     /label=promoter -10 of Plac
                     /note="promoter -10 of Plac"
     promoter        complement(5636..5666)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     misc_feature    complement(5661..5666)
                     /label=promoter -30 of Plac
                     /note="promoter -30 of Plac"
     protein_bind    5681..5702
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    complement(5975..6593)
                     /label=rep(pMB1)
                     /note="rep(pMB1)"
     rep_origin      complement(5990..6578)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6749..7609)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="AEA29844.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(6749..7609)
                     /gene="bla"
                     /label=bla
     promoter        complement(7610..22)
                     /gene="bla"
                     /label=AmpR promoter