LOCUS       Exported                7082 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pCon5, complete sequence.
ACCESSION   JF276395
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7082)
  AUTHORS   Raja Abd Rahman RNZ., Wong CF, Salleh AB, Basri M.
  TITLE     Expression vector of pCon5
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7082)
  AUTHORS   Raja Abd Rahman RNZ., Wong CF, Salleh AB, Basri M.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-JAN-2011) Department of Microbiology, Universiti Putra
            Malaysia, Faculty of Biotechnology and Biomolecular Sciences, UPM 
            Serdang, Selangor 43400, Malaysia
REFERENCE   3  (bases 1 to 7082)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7082
                     /organism="Expression vector pCon5"
                     /mol_type="other DNA"
                     /db_xref="taxon:999511"
     misc_feature    complement(47..665)
                     /label=rep(pMB1)
                     /note="rep(pMB1)"
     rep_origin      complement(62..650)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(821..1681)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="AEA29839.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(821..1681)
                     /gene="bla"
                     /label=bla
     promoter        complement(1682..1786)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1977..2054
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             2046..3137
                     /codon_start=1
                     /gene="lacIq"
                     /product="LacIq"
                     /label=lacIq
                     /note="LacI repressor"
                     /protein_id="AEA29840.1"
                     /translation="MVNVKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAM
                     AELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVER
                     SGVEACKAAGHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII
                     FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGD
                     WSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTED
                     SSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTA
                     SPRALADSLMQLARQVSRLESGQ"
     gene            2046..3137
                     /gene="lacIq"
                     /label=lacIq
     CDS             2055..3137
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAGHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     CDS             complement(3327..4823)
                     /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="AEA29841.1"
                     /translation="MKKVSTLDLLFVAIMGVSPAALAADLIDVSKLPSKAAQGAPGPVT
                     LQAAVGAGGADELKAIRSTTLPNGKQVTRYEQFHNGVRVVGEAITEVKGPGKSVAAQRS
                     GHFVANIAADLPGSTTAAVSAEQVLAQAKSLKAQGRKTENDKVELVIRLGESNIAQLVY
                     NVSYLIPGEGLSRPHFVIDAKTGEVLDQWEGLAHAEAGGPGGNQKIGKYTYGSDYGPLI
                     VNDRCEMDDGNVITVDMNGSTDDSKTTPFRFACPTNTYKQVNGAYSPLNDAHFFGGVVF
                     KLYRDWFGTSPLTHKLYMKVHYGRSVENAYWGGTAMLFGDGATMFYPLVSLDVAAHEVS
                     HGFTEQNSGLIYRGQSGGMNEAFSDMAGEAAEFYMRGKNDFLIGYDIKKGSGALRYMDQ
                     PSRDGRSIDNASQYYNGIDVHHSSGVYNRAFYLLANSPGWDTRKAFEVFVDANRYYWTA
                     TSNYNSGACGVIRSAQNRNYSAADVTRAFSTVGVTCPSAL"
     misc_feature    complement(4869..4897)
                     /label=operator O3 of PT7(A1/O4/O3)
                     /note="operator O3 of PT7(A1/O4/O3)"
     protein_bind    4879..4895
                     /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(4905..4910)
                     /label=promoter -10 of PT7(A1/O4/O3)
                     /note="promoter -10 of PT7(A1/O4/O3)"
     misc_feature    complement(4911..4927)
                     /label=operator O4 of PT7(A1/O4/O3)
                     /note="operator O4 of PT7(A1/O4/O3)"
     protein_bind    4912..4928
                     /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(4928..4933)
                     /label=promoter -33 of PT7(A1/O4/O3)
                     /note="promoter -33 of PT7(A1/O4/O3)"
     misc_feature    5111..6892
                     /label=SF
                     /note="SF"
     rep_origin      5662..6013
                     /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             6027..6860
                     /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     7021..7037
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"