LOCUS       Exported                6706 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pMAL-pIII, complete sequence.
ACCESSION   AF031088
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6706)
  AUTHORS   Zwick MB, Bonnycastle LL, Noren KA, Venturini S, Leong E, Barbas CF 
            III, Noren CJ, Scott JK.
  TITLE     The maltose-binding protein as a scaffold for monovalent display of 
            peptides derived from phage libraries
  JOURNAL   Anal. Biochem. 264 (1), 87-97 (1998)
  PUBMED    9784192
REFERENCE   2  (bases 1 to 6706)
  AUTHORS   Noren KA, Noren CJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-OCT-1997) New England Biolabs, 32 Tozer Road, Beverly,
            MA 01915, USA
REFERENCE   3  (bases 1 to 6706)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6706
                     /organism="Shuttle vector pMAL-pIII"
                     /mol_type="genomic DNA"
                     /note="derived from commercially available vector pMal-p2 
                     (New England Biolabs)"
                     /db_xref="taxon:69327"
     promoter        3..80
                     /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."
     gene            15..1163
                     /gene="lacI"
                     /label=lacI
                     /note="E. coli gene"
     regulatory      15..46
                     /regulatory_class="promoter"
                     /gene="lacI"
     CDS             81..1163
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="transcriptional repressor"
                     /protein_id="AAB86558.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    1176..1197
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1406..1434
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     regulatory      1406..1433
                     /regulatory_class="promoter"
                     /note="Ptac promoter"
     protein_bind    1442..1458
                     /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)."
     CDS             1528..2976
                     /codon_start=1
                     /gene="malE-lacZa fusion"
                     /product="maltose binding protein-lacZ alpha peptide fusion
                     protein precursor"
                     /label=malE-lacZa fusion
                     /note="MBP-lacZa; MBP modified to include phage M13 
                     secretion signal sequence"
                     /protein_id="AAB86559.1"
                     /translation="MKKLLFAIPLVVPFYSHSADMKIEEGKLVIWINGDKGYNGLAEVG
                     KKFEKDTGIKVTVEHPDKLEEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDK
                     AFQDKLYPFTWDAVRYNGKLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAK
                     GKSALMFNLQEPYFTWPLIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKN
                     KHMNADTDYSIAEAAFNKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVG
                     VLSAGINAASPNKELAKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAA
                     TMENAQKGEIMPNIPQMSAFWYAVRTAVINAASGRQTVDEALKDAQTNSSSNNNNNNNN
                     NNLGIEGRISEFGSSRVDLQASLALAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEE
                     ARTDRPSQQLRSLNGEWQLGCFGG"
     gene            1528..2976
                     /gene="malE-lacZa fusion"
                     /label=malE-lacZa fusion
                     /note="E. coli gene fusion"
     sig_peptide     1528..1581
                     /gene="malE-lacZa fusion"
                     /note="M13 secretion signal sequence"
     mat_peptide     1582..2973
                     /gene="malE-lacZa fusion"
                     /product="maltose binding protein-lacZ alpha peptide fusion
                     protein"
     CDS             1588..2688
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     CDS             2737..2748
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2796..2812)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      3161..3247
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3339..3366
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3386..3477
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3478..4338
                     /codon_start=1
                     /gene="bla"
                     /product="RTEM-1 beta-lactamase"
                     /label=bla
                     /note="penicillinase"
                     /protein_id="AAB86560.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            3478..4338
                     /gene="bla"
                     /label=bla
                     /note="R-plasmid ampicillin resistance gene"
     rep_origin      complement(4380..4893)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      4381..4893
                     /standard_name="M13 origin (+ -)"
                     /label=M13 origin (+ -)
     rep_origin      5004..5592
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    5778..5918
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(6020..6211)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"