LOCUS FJ010196 9991bp DNA circular SYN 11-AUG-2008 DEFINITION Expression vector pKLMF-EK, complete sequence. ACCESSION FJ010196 VERSION FJ010196.1 KEYWORDS . SOURCE Expression vector pKLMF-EK ORGANISM Expression vector pKLMF-EK other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 9991) AUTHORS Ganatra,M.B., Read,J.D., Colussi,P.A. and Taron,C.H. TITLE Vectors for intracellular expression of maltose binding protein fusions in the yeast Kluyveromyces lactis JOURNAL Unpublished REFERENCE 2 (bases 1 to 9991) AUTHORS Ganatra,M.B., Read,J.D., Colussi,P.A. and Taron,C.H. TITLE Direct Submission JOURNAL Submitted (11-AUG-2008) Gene Expression Division, New England Biolabs, 240 County Road, Ipswich, MA 01938 FEATURES Location/Qualifiers CDS 13..1113 /gene=" malE" /translation="MKTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKL EEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNG KLIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWP LIAADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAF NKGETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKEL AKEFLENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNI PQMSAFWYAVRTAVINAASGRQTVDEALKDAQT" /product=" maltose binding protein (MBP)" CDS 1114..1140 /note=" spacer, Enterokinase site" /translation="NSSSDDDDK" misc_feature 1141..1237 /note="multiple cloning site" terminator 1254..1837 /note="Kluyveromyces lactis LAC4 transcription terminator" promoter 1893..2596 /note="Saccharomyces cerevisiae ADH1 promoter" gene 2597..4243 /gene="amdS" CDS 2597..4243 /gene="amdS" /note="confers ability to process acetamide for selection in Kluyveromyces lactis" /codon_start=1 /transl_table=11 /product="acetamidase" /protein_id="ABW72883.1" /db_xref="GI:158536734" /translation="MPQSWEELAADKRARLAKTIPDEWKVQTLPAEDSVIDFPKKSGI LSEAELKITEASAADLVSKLAAGELTSVEVTLAFCKRAAIAQQLTNCAHEFFPDAALA QARELDEYYAKHKRPVGPLHGLPISLKDQLRVKGYETSMGYISWLNKYDEGDSVLTTM LRKAGAVFYVKTSVPQTLMVCETVNNIIGRTVNPRNKNWSCGGSSGGEGAIVGIRGGV IGVGTDIGGSIRVPAAFNFLYGLRPSHGRLPYAKMANSMEGQETVHSVVGPITHSVED LRLFTKSVLGQEPWKYDSKVIPMPWRQSESDIIASKIKNGGLNIGYYNFDGNVLPHPP ILRGVETTVAALAKAGHTVTPWTPYKHDFGHDLISHIYAADGSADVMRDISASGEPAI PNIKDLLNPNIKAVNMNELWDTHLQKWNYQMEYLEKWREAEEKAGKELDAIIAPITPT AAVRHDQFRYYGYASVINLLDFTSVVVPVTFADKNIDKKNESFKAVSELDALVQEEYD PEAYHGAPVAVQVIGRRLSEERTLAIAEEVGKLLGNVVTP" promoter 4951..5531 /note="modified Kluyveromyces lactis LAC4 promoter 5' end" gene complement(6745..7605) /gene="bla" CDS complement(6745..7605) /gene="bla" /note="confers ampicillin resistance in Escherichia coli" /codon_start=1 /transl_table=11 /product="beta-lactamase" /protein_id="ABW72884.1" /db_xref="GI:158536735" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" promoter 8359..9991 /note="modified Kluyveromyces lactis LAC4 promoter 3' end" BASE COUNT 2673 a 2354 c 2392 g 2572 t ORIGIN 1 aagcttgcca ccatgaaaac tgaagaaggt aaactggtaa tctggattaa cggcgataaa 61 ggctataacg gtctcgctga agtcggtaag aaattcgaga aagataccgg aattaaagtc 121 accgttgagc atccggataa actggaagag aaattcccac aggttgcggc aactggcgat 181 ggccctgaca ttatcttctg ggcacacgac cgctttggtg gctacgctca atctggcctg 241 ttggctgaaa tcaccccgga caaagcgttc caggacaagc tgtatccgtt tacctgggat 301 gccgtacgtt acaacggcaa gctgattgct tacccgatcg ctgttgaagc gttatcgctg 361 atttataaca aagatctgct gccgaacccg ccaaaaacct gggaagagat cccggcgctg 421 gataaagaac tgaaagcgaa aggtaagagc gcgctgatgt tcaacctgca agaaccgtac 481 ttcacctggc cgctgattgc tgctgacggg ggttatgcgt tcaagtatga aaacggcaag 541 tacgacatta aagacgtggg cgtggataac gctggcgcga aagcgggtct gaccttcctg 601 gttgacctga ttaaaaacaa acacatgaat gcagacaccg attactccat cgcagaagct 661 gcctttaata aaggcgaaac agcgatgacc atcaacggcc cgtgggcatg gtccaacatc 721 gacaccagca aagtgaatta tggtgtaacg gtactgccga ccttcaaggg tcaaccatcc 781 aaaccgttcg ttggcgtgct gagcgcaggt attaacgccg ccagtccgaa caaagagctg 841 gcaaaagagt tcctcgaaaa ctatctgctg actgatgaag gtctggaagc ggttaataaa 901 gacaaaccgc tgggtgccgt agcgctgaag tcttacgagg aagagttggc gaaagatcca 961 cgtattgccg ccactatgga aaacgcccag aaaggtgaaa tcatgccgaa catcccgcag 1021 atgtccgctt tctggtatgc cgtgcgtact gcggtgatca acgccgccag cggtcgtcag 1081 actgtcgatg aagccctgaa agacgcgcag actaattcga gctcggatga cgatgacaag 1141 ctcgagaaaa gagaggctga agctagaaga gctcatatgt ccatgggcgg ccgcgatatc 1201 gtcgacggat ccgaattccc tgcaggtaat taaataaagg ccttgaatcg agaatttata 1261 cttagataag tatgtactta caggtatatt tctatgagat actgatgtat acatgcatga 1321 taatatttaa acggttatta gtgccgattg tcttgtgcga taatgacgtt cctatcaaag 1381 caatacactt accacctatt acatgggcca agaaaatatt ttcgaacttg tttagaatat 1441 tagcacagag tatatgatgt tatccgttag attatgcatg attcattcct acaacttttt 1501 cgtagcataa ggattaatta cttggatgcc aataaaaaaa aaaaacatcg agaaaatttc 1561 agcatgctca gaaacaattg cagtgtatca aagtaaaaaa aagattttca ctacatgttc 1621 cttttgaaga aagaaaatca tggaacatta gatttacaaa aatttaacca ccgctgatta 1681 acgattagac cgttaagcgc acaacaggtt attagtacag agaaagcatt ctgtggtgtt 1741 gccccggact ttcttttgcg acataggtaa atcgaatacc atcatactat cttttccaat 1801 gactccctaa agaaagactc ttcttcgatg ttgtatacgt tggagcatag ggcaagaatt 1861 gtggcttgag atcatccttt tgttgtttcc gggtgtacaa tatggacttc ctcttttctg 1921 gcaaccaaac ccatacatcg ggattcctat aataccttcg ttggtctccc taacatgtag 1981 gtggcggagg ggagatatac aatagaacag ataccagaca agacataatg ggctaaacaa 2041 gactacacca attacactgc ctcattgatg gtggtacata acgaactaat actgtagccc 2101 tagacttgat agccatcatc atatcgaagt ttcactaccc tttttccatt tgccatctat 2161 tgaagtaata ataggcgcat gcaacttctt ttcttttttt ttcttttctc tctcccccgt 2221 tgttgtctca ccatatccgc aatgacaaaa aaatgatgga agacactaaa ggaaaaaatt 2281 aacgacaaag acagcaccaa cagatgtcgt tgttccagag ctgatgaggg gtatctcgaa 2341 gcacacgaaa ctttttcctt ccttcattca cgcacactac tctctaatga gcaacggtat 2401 acggccttcc ttccagttac ttgaatttga aataaaaaaa agtttgctgt cttgctatca 2461 agtataaata gacctgcaat tattaatctt ttgtttcctc gtcattgttc tcgttccctt 2521 tcttccttgt ttctttttct gcacaatatt tcaagctata ccaagcatac aatcaagcaa 2581 ttccagatct gccaccatgc ctcaatcctg ggaagaactg gccgctgata agcgcgcccg 2641 cctcgcaaaa accatccctg atgaatggaa agtccagacg ctgcctgcgg aagacagcgt 2701 tattgatttc ccaaagaaat cgggcatcct ttcagaggcc gaactgaaga tcacagaggc 2761 ttccgctgcg gatcttgtgt ccaagctggc ggccggagag ttgacctcgg tggaagttac 2821 gctagcattc tgtaaacggg cagcaatcgc ccagcagtta acaaactgcg cccacgagtt 2881 cttccctgac gccgctctcg cgcaggcaag ggaactcgat gaatactacg caaagcacaa 2941 gagacccgtt ggtccacttc atggcctccc catctctctc aaagaccagc ttcgagtcaa 3001 gggctacgaa acatcaatgg gctacatctc atggctaaac aagtacgacg aaggggactc 3061 ggttctgaca accatgctcc gcaaagccgg tgccgtcttc tacgtcaaga cctctgtccc 3121 gcagaccctg atggtctgcg agacagtcaa caacatcatc gggcgcaccg tcaacccacg 3181 caacaagaac tggtcgtgcg gcggcagttc tggtggtgag ggtgcgatcg ttgggattcg 3241 tggtggcgtc atcggtgtag gaacggacat cggtggctcg attcgagtgc cggccgcgtt 3301 caacttcctg tacggtctaa ggccgagtca tgggcggctg ccgtatgcaa agatggcgaa 3361 cagcatggag ggtcaggaga cggtgcacag cgttgtcggg ccgattacgc actctgttga 3421 ggacctccgc ctcttcacca aatccgtcct cggtcaggag ccttggaaat acgactccaa 3481 ggtcatcccc atgccctggc gccagtccga gtcggacatt attgcctcca agatcaagaa 3541 cggcgggctc aatatcggct actacaactt cgacggcaat gtccttccac accctcctat 3601 cctgcgcggc gtggaaacta ccgtcgccgc actcgccaaa gccggtcaca ccgtgacccc 3661 gtggacgcca tacaagcacg atttcggcca cgatctcatc tcccatatct acgcggctga 3721 cggcagcgcc gacgtaatgc gcgacatcag tgcatccggc gagccggcga ttccaaatat 3781 caaagaccta ctgaacccga acatcaaagc tgttaacatg aacgagctct gggacacgca 3841 tctccagaag tggaattacc agatggagta ccttgagaaa tggcgggagg ctgaagaaaa 3901 ggccgggaag gaactggacg ccatcatcgc gccgattacg cctaccgctg cggtacggca 3961 tgaccagttc cggtactatg ggtatgcctc tgtgatcaac ctgctggatt tcacgagcgt 4021 ggttgttccg gttacctttg cggataagaa catcgataag aagaatgaga gtttcaaggc 4081 ggttagtgag cttgatgccc tcgtgcagga agagtatgat ccggaggcgt atcatggggc 4141 accggttgca gtgcaggtta tcggacggag actcagtgaa gagaggacgt tggcgattgc 4201 agaggaagtg gggaagttgc tgggaaatgt ggtgactcca tagcccgggg ggggctcgat 4261 cccctcgcga gttggttcag ctgctgcctg aggctggacg acctcgcgga gttctaccgg 4321 cagtgcaaat ccgtcggcat ccaggaaacc agcagcggct atccgcgcat ccatgccccc 4381 gaactgcagg agtggggagg cacgatggcc gctttggtcg atctagatta cgtggaagaa 4441 aggtagtaaa agtagtagta taagtagtaa aaagaggtaa aaagagaaaa ccggctacat 4501 actagagaag cacgtacaca aaaactcata ggcacttcat catacgacag tttcttgatg 4561 cattataata gtgtattaga tattttcaga aatatgcata gaacctcctc ttgcctttac 4621 tttttataca tagaacattg gcagatttac ttacactact ttgtttctac gccatttctt 4681 ttgttttcaa cacttagaca agttgttgag aaccggacta ctaaaaagca atgttcccac 4741 tgaaaatcat gtacctgcag cataataacc ccctaattct gcatcgatcc agtatgtttt 4801 tttttctcta ctcattttta cctgaagata gagcttctaa aacaaaaaaa atcagtgatt 4861 acatgcatat tgtgtgttct agtaaccaaa ggaaaggaac agatagataa aattccgaga 4921 ctgtcaaatt aggttttttt cttttttttt ggcgggagtc agtgggccga aatatgttct 4981 tggcctagaa cttaatctgg tttgatcatg ccaatacttg cctgagtgcc cgactttttg 5041 cccaccctct tgccttctgt catccttcaa aacccacctg ttttccagcc gtatcttcgc 5101 tcgcatctac acatactgtg ccatatcttg tgtgtagccg gacgtgacta tgaccaaaaa 5161 caaacaagga gaactgttcg ccgatttgta acactcctgc atccatccaa gtgggtatgc 5221 gctatgcaat gttaagctag gtcaggtcag accaggtcca aggacagcaa cttgactgta 5281 tgcaaccttt accatctttg cacagaacat acttgtagct agctagttac acttatggac 5341 cgaaaaggca ccccaccatg tctgtccggc tttagagtac ggccgcagac cgctgatttg 5401 ccttgccaag cagtagtcac aatgcatcgc atgagcacac gggcacgggc acgggcacag 5461 gaaccattgg caaaaatacc agatacacta taccgacgta tatcaagccc aagtttaaaa 5521 ttcctaaatt tccgcggcta cttttcaatt ccctatagtg agtcgtatta aattcgtaat 5581 catgtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 5641 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 5701 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 5761 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 5821 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 5881 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 5941 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 6001 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 6061 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 6121 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 6181 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 6241 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 6301 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 6361 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 6421 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 6481 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 6541 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 6601 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 6661 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 6721 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 6781 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 6841 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 6901 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 6961 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 7021 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 7081 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 7141 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 7201 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 7261 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 7321 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 7381 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 7441 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 7501 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 7561 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 7621 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 7681 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc 7741 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac 7801 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt 7861 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc 7921 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc 7981 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact 8041 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg 8101 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc 8161 gaattttaac aaaatattaa cgcttacaat ttccattcgc cattcaggct gcgcaactgt 8221 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt 8281 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg 8341 acggccagtg ccaagctccc gcggggatcg actcataaaa tagtaacctt ctaatgcgta 8401 tctattgact accaaccatt agtgtggttg cagaaggcgg aattttccct tcttcgaatt 8461 tagcttgctt tttcattttt tattttccat ttttcagttt ttgtttgtgt cgaatttagc 8521 cagttgcttc tccaagatga aaaaaacccc tgcgcagttt ctgtgctgca agatcctaat 8581 cgacttttcc accccccaca aaagtaaatg ttcttttgtt acattcgcgt gggtagctag 8641 ctccccgaat cttcaaagga cttagggact gcactacatc agagtgtgtt cacctggttt 8701 gctgcctggt ttgaaagaaa agagcaggga actcgcgggt tcccggcgaa taatcatgcg 8761 atagtccttt ggccttccaa gtcgcatgta gagtagacaa cagacaggga gggcaggaag 8821 gatctttcac tgagatcctg tatcttgttg ggtaagtcgg atgaaagggg aatcgtatga 8881 gattggagag gatgcggaag aggtaacgcc ttttgttaac ttgtttaatt attatggggc 8941 aggcgagagg gggaggaatg tatgtgtgtg aggcgggcga gacggagcca tccaggccag 9001 gtagaaatag agaaagccga atgttagaca atatggcagc gtagtagagt aggtaggtag 9061 gcaagtactg ctagcaaaga ggagaagggt aagctcactc ttcgcattcc acaccgttag 9121 tgtgtcagtt tgaacaaaaa aacaatcatc ataccaattg atggactgtg gactggcttt 9181 tggaacggct tttcggactg cgattattcg tgaggaatca aggtaggaat ttggtcatat 9241 ttacggacaa cagtgggtga ttcccatatc gagtaggaaa acgagatcat ggtatcctca 9301 gatatgttgc ggaaatctgt tcaccgcaaa gttcagggtg ctctggtggg tttcggttgg 9361 tctttgcttt gcttctccct tgtcttgcat gttaataata gcctagcctg tgagccgaaa 9421 cttagggtag gcttagtgtt ggaacgtaca tatctatcac gttgacttgg tttaaccagg 9481 cgacctggta gccagccata cccacacacg ttttttgtat cttcagtata gttgtgaaaa 9541 gtgtagcgga aatttgtggt ccgagcaaca gcgtcttttt ctagtagtgc ggtcggttac 9601 ttggttgaca ttggtatttg gactttgttg ctacaccatt cactacttga agtcgagtgt 9661 gaagggtatg atttctagtg gtgaacacct ttagttacgt aatgttttca ttgctgtttt 9721 acttgagatt tcgattgaga aaaaggtatt taatagctcg aatcaatgtg agaacagaga 9781 gaagatgttc ttccctaact cgaaaggtat atgaggcttg tgtttcttag gagaattatt 9841 attcttttgt tatgttgcgc ttgtagttgg aaaaggtgaa gagacaaaag ctggaattgt 9901 gagcggataa caagctcaac acttgaaatt taggaaagag cagaatttgg caaaaaaaat 9961 aaaaaaaaaa taaacacaca tactcatcga g //