LOCUS FJ010197 9988bp DNA circular SYN 11-AUG-2008 DEFINITION Expression vector pKLMF-FX, complete sequence. ACCESSION FJ010197 VERSION FJ010197.1 KEYWORDS . SOURCE Expression vector pKLMF-FX ORGANISM Expression vector pKLMF-FX other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 9988) 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TITLE Vectors for intracellular expression of maltose binding protein fusions in the yeast Kluyveromyces lactis JOURNAL Unpublished REFERENCE 2 (bases 1 to 9988) AUTHORS Ganatra,M.B., Read,J.D., Colussi,P.A. and Taron,C.H. TITLE Direct Submission JOURNAL Submitted (11-AUG-2008) Gene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..1137 /note=" spacer, factor Xa site" /translation="NSSSIEGR" misc_feature 1138..1234 /note="multiple cloning site" terminator 1251..1834 /note="Kluyveromyces lactis LAC4 transcription terminator" promoter 1890..2593 /note="Saccharomyces cerevisiae ADH1 promoter" gene 2594..4240 /gene="amdS" CDS 2594..4240 /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 4948..5528 /note="modified Kluyveromyces lactis LAC4 promoter 5' end" gene complement(6742..7602) /gene="bla" CDS complement(6742..7602) /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 8356..9988 /note="modified Kluyveromyces lactis LAC4 promoter 3' end" BASE COUNT 2671 a 2353 c 2393 g 2571 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 gctcgatcga gggaaggctc 1141 gagaaaagag aggctgaagc tagaagagct catatgtcca tgggcggccg cgatatcgtc 1201 gacggatccg aattccctgc aggtaattaa ataaaggcct tgaatcgaga atttatactt 1261 agataagtat gtacttacag gtatatttct atgagatact gatgtataca tgcatgataa 1321 tatttaaacg gttattagtg ccgattgtct tgtgcgataa tgacgttcct atcaaagcaa 1381 tacacttacc acctattaca tgggccaaga aaatattttc gaacttgttt agaatattag 1441 cacagagtat atgatgttat ccgttagatt atgcatgatt cattcctaca actttttcgt 1501 agcataagga ttaattactt ggatgccaat aaaaaaaaaa aacatcgaga aaatttcagc 1561 atgctcagaa acaattgcag tgtatcaaag taaaaaaaag attttcacta catgttcctt 1621 ttgaagaaag aaaatcatgg aacattagat ttacaaaaat ttaaccaccg ctgattaacg 1681 attagaccgt taagcgcaca acaggttatt agtacagaga aagcattctg tggtgttgcc 1741 ccggactttc ttttgcgaca taggtaaatc gaataccatc atactatctt ttccaatgac 1801 tccctaaaga aagactcttc ttcgatgttg tatacgttgg agcatagggc aagaattgtg 1861 gcttgagatc atccttttgt tgtttccggg tgtacaatat ggacttcctc ttttctggca 1921 accaaaccca tacatcggga ttcctataat accttcgttg gtctccctaa catgtaggtg 1981 gcggagggga gatatacaat agaacagata ccagacaaga cataatgggc taaacaagac 2041 tacaccaatt acactgcctc attgatggtg gtacataacg aactaatact gtagccctag 2101 acttgatagc catcatcata tcgaagtttc actacccttt ttccatttgc catctattga 2161 agtaataata ggcgcatgca acttcttttc tttttttttc ttttctctct cccccgttgt 2221 tgtctcacca tatccgcaat gacaaaaaaa tgatggaaga cactaaagga aaaaattaac 2281 gacaaagaca gcaccaacag atgtcgttgt tccagagctg atgaggggta tctcgaagca 2341 cacgaaactt tttccttcct tcattcacgc acactactct ctaatgagca acggtatacg 2401 gccttccttc cagttacttg aatttgaaat aaaaaaaagt ttgctgtctt gctatcaagt 2461 ataaatagac ctgcaattat taatcttttg tttcctcgtc attgttctcg ttccctttct 2521 tccttgtttc tttttctgca caatatttca agctatacca agcatacaat caagcaattc 2581 cagatctgcc accatgcctc aatcctggga agaactggcc gctgataagc gcgcccgcct 2641 cgcaaaaacc atccctgatg aatggaaagt ccagacgctg cctgcggaag acagcgttat 2701 tgatttccca aagaaatcgg gcatcctttc agaggccgaa ctgaagatca cagaggcttc 2761 cgctgcggat cttgtgtcca agctggcggc cggagagttg acctcggtgg aagttacgct 2821 agcattctgt aaacgggcag caatcgccca gcagttaaca aactgcgccc acgagttctt 2881 ccctgacgcc gctctcgcgc aggcaaggga actcgatgaa tactacgcaa agcacaagag 2941 acccgttggt ccacttcatg gcctccccat ctctctcaaa gaccagcttc gagtcaaggg 3001 ctacgaaaca tcaatgggct acatctcatg gctaaacaag tacgacgaag gggactcggt 3061 tctgacaacc atgctccgca aagccggtgc cgtcttctac gtcaagacct ctgtcccgca 3121 gaccctgatg gtctgcgaga cagtcaacaa catcatcggg cgcaccgtca acccacgcaa 3181 caagaactgg tcgtgcggcg gcagttctgg tggtgagggt gcgatcgttg ggattcgtgg 3241 tggcgtcatc ggtgtaggaa cggacatcgg tggctcgatt cgagtgccgg ccgcgttcaa 3301 cttcctgtac ggtctaaggc cgagtcatgg gcggctgccg tatgcaaaga tggcgaacag 3361 catggagggt caggagacgg tgcacagcgt tgtcgggccg attacgcact ctgttgagga 3421 cctccgcctc ttcaccaaat ccgtcctcgg tcaggagcct tggaaatacg actccaaggt 3481 catccccatg ccctggcgcc agtccgagtc ggacattatt gcctccaaga tcaagaacgg 3541 cgggctcaat atcggctact acaacttcga cggcaatgtc cttccacacc ctcctatcct 3601 gcgcggcgtg gaaactaccg tcgccgcact cgccaaagcc ggtcacaccg tgaccccgtg 3661 gacgccatac aagcacgatt tcggccacga tctcatctcc catatctacg cggctgacgg 3721 cagcgccgac gtaatgcgcg acatcagtgc atccggcgag ccggcgattc caaatatcaa 3781 agacctactg aacccgaaca tcaaagctgt taacatgaac gagctctggg acacgcatct 3841 ccagaagtgg aattaccaga tggagtacct tgagaaatgg cgggaggctg aagaaaaggc 3901 cgggaaggaa ctggacgcca tcatcgcgcc gattacgcct accgctgcgg tacggcatga 3961 ccagttccgg tactatgggt atgcctctgt gatcaacctg ctggatttca cgagcgtggt 4021 tgttccggtt acctttgcgg ataagaacat cgataagaag aatgagagtt tcaaggcggt 4081 tagtgagctt gatgccctcg tgcaggaaga gtatgatccg gaggcgtatc atggggcacc 4141 ggttgcagtg caggttatcg gacggagact cagtgaagag aggacgttgg cgattgcaga 4201 ggaagtgggg aagttgctgg gaaatgtggt gactccatag cccggggggg gctcgatccc 4261 ctcgcgagtt ggttcagctg ctgcctgagg ctggacgacc tcgcggagtt ctaccggcag 4321 tgcaaatccg tcggcatcca ggaaaccagc agcggctatc cgcgcatcca tgcccccgaa 4381 ctgcaggagt ggggaggcac gatggccgct ttggtcgatc tagattacgt ggaagaaagg 4441 tagtaaaagt agtagtataa gtagtaaaaa gaggtaaaaa gagaaaaccg gctacatact 4501 agagaagcac gtacacaaaa actcataggc acttcatcat acgacagttt cttgatgcat 4561 tataatagtg tattagatat tttcagaaat atgcatagaa cctcctcttg cctttacttt 4621 ttatacatag aacattggca gatttactta cactactttg tttctacgcc atttcttttg 4681 ttttcaacac ttagacaagt tgttgagaac cggactacta aaaagcaatg ttcccactga 4741 aaatcatgta cctgcagcat aataaccccc taattctgca tcgatccagt atgttttttt 4801 ttctctactc atttttacct gaagatagag cttctaaaac aaaaaaaatc agtgattaca 4861 tgcatattgt gtgttctagt aaccaaagga aaggaacaga tagataaaat tccgagactg 4921 tcaaattagg tttttttctt tttttttggc gggagtcagt gggccgaaat atgttcttgg 4981 cctagaactt aatctggttt gatcatgcca atacttgcct gagtgcccga ctttttgccc 5041 accctcttgc cttctgtcat ccttcaaaac ccacctgttt tccagccgta tcttcgctcg 5101 catctacaca tactgtgcca tatcttgtgt gtagccggac gtgactatga ccaaaaacaa 5161 acaaggagaa ctgttcgccg atttgtaaca ctcctgcatc catccaagtg ggtatgcgct 5221 atgcaatgtt aagctaggtc aggtcagacc aggtccaagg acagcaactt gactgtatgc 5281 aacctttacc atctttgcac agaacatact tgtagctagc tagttacact tatggaccga 5341 aaaggcaccc caccatgtct gtccggcttt agagtacggc cgcagaccgc tgatttgcct 5401 tgccaagcag tagtcacaat gcatcgcatg agcacacggg cacgggcacg ggcacaggaa 5461 ccattggcaa aaataccaga tacactatac cgacgtatat caagcccaag tttaaaattc 5521 ctaaatttcc gcggctactt ttcaattccc tatagtgagt cgtattaaat tcgtaatcat 5581 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 5641 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc 5701 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 5761 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 5821 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 5881 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 5941 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 6001 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 6061 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 6121 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 6181 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 6241 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 6301 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 6361 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 6421 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 6481 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 6541 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 6601 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 6661 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 6721 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 6781 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 6841 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 6901 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 6961 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 7021 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 7081 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 7141 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 7201 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 7261 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 7321 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 7381 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 7441 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 7501 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 7561 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 7621 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 7681 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc 7741 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact 7801 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc 7861 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt 7921 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc 7981 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt 8041 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat 8101 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa 8161 ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg caactgttgg 8221 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct 8281 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg 8341 gccagtgcca agctcccgcg gggatcgact cataaaatag taaccttcta atgcgtatct 8401 attgactacc aaccattagt gtggttgcag aaggcggaat tttcccttct tcgaatttag 8461 cttgcttttt cattttttat tttccatttt tcagtttttg tttgtgtcga atttagccag 8521 ttgcttctcc aagatgaaaa aaacccctgc gcagtttctg tgctgcaaga tcctaatcga 8581 cttttccacc ccccacaaaa gtaaatgttc ttttgttaca ttcgcgtggg tagctagctc 8641 cccgaatctt caaaggactt agggactgca ctacatcaga gtgtgttcac ctggtttgct 8701 gcctggtttg aaagaaaaga gcagggaact cgcgggttcc cggcgaataa tcatgcgata 8761 gtcctttggc cttccaagtc gcatgtagag tagacaacag acagggaggg caggaaggat 8821 ctttcactga gatcctgtat cttgttgggt aagtcggatg aaaggggaat cgtatgagat 8881 tggagaggat gcggaagagg taacgccttt tgttaacttg tttaattatt atggggcagg 8941 cgagaggggg aggaatgtat gtgtgtgagg cgggcgagac ggagccatcc aggccaggta 9001 gaaatagaga aagccgaatg ttagacaata tggcagcgta gtagagtagg taggtaggca 9061 agtactgcta gcaaagagga gaagggtaag ctcactcttc gcattccaca ccgttagtgt 9121 gtcagtttga acaaaaaaac aatcatcata ccaattgatg gactgtggac tggcttttgg 9181 aacggctttt cggactgcga ttattcgtga ggaatcaagg taggaatttg gtcatattta 9241 cggacaacag tgggtgattc ccatatcgag taggaaaacg agatcatggt atcctcagat 9301 atgttgcgga aatctgttca ccgcaaagtt cagggtgctc tggtgggttt cggttggtct 9361 ttgctttgct tctcccttgt cttgcatgtt aataatagcc tagcctgtga gccgaaactt 9421 agggtaggct tagtgttgga acgtacatat ctatcacgtt gacttggttt aaccaggcga 9481 cctggtagcc agccataccc acacacgttt tttgtatctt cagtatagtt gtgaaaagtg 9541 tagcggaaat ttgtggtccg agcaacagcg tctttttcta gtagtgcggt cggttacttg 9601 gttgacattg gtatttggac tttgttgcta caccattcac tacttgaagt cgagtgtgaa 9661 gggtatgatt tctagtggtg aacaccttta gttacgtaat gttttcattg ctgttttact 9721 tgagatttcg attgagaaaa aggtatttaa tagctcgaat caatgtgaga acagagagaa 9781 gatgttcttc cctaactcga aaggtatatg aggcttgtgt ttcttaggag aattattatt 9841 cttttgttat gttgcgcttg tagttggaaa aggtgaagag acaaaagctg gaattgtgag 9901 cggataacaa gctcaacact tgaaatttag gaaagagcag aatttggcaa aaaaaataaa 9961 aaaaaaataa acacacatac tcatcgag //