LOCUS EU196354 9107 bp DNA circular SYN 14-AUG-2008 DEFINITION Expression vector pKLAC2, complete sequence. ACCESSION EU196354 VERSION EU196354.2 GI:196122458 KEYWORDS . SOURCE Expression vector pKLAC2 ORGANISM Expression vector pKLAC2 other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 9107) AUTHORS Colussi,P.A., Read,J.D. and Taron,C.H. TITLE An integrative expression vector for the yeast Kluyveromyces lactis JOURNAL Unpublished REFERENCE 2 (bases 1 to 9107) AUTHORS Colussi,P.A., Read,J.D. and Taron,C.H. TITLE Direct Submission JOURNAL Submitted (08-OCT-2007) Gene Expression Division, New England Biolabs, 240 County Road, Ipswich, MA 01938, USA REFERENCE 3 (bases 1 to 9107) AUTHORS Colussi,P.A., Read,J.D. and Taron,C.H. TITLE Direct Submission JOURNAL Submitted (14-AUG-2008) Gene Expression Division, New England Biolabs, 240 County Road, Ipswich, MA 01938, USA REMARK Sequence update by submitter COMMENT On Aug 14, 2008 this sequence version replaced gi:158536733. FEATURES Location/Qualifiers source 1..9107 /organism="Expression vector pKLAC2" /mol_type="other DNA" /db_xref="taxon:478548" misc_feature 14..353 /note="Kluyveromyces lactis alpha mating factor pre-pro secretion leader" misc_feature 290..353 /note="multiple cloning site" terminator 370..953 /note="Kluyveromyces lactis LAC4 transcription terminator" promoter 1009..1712 /note="Saccharomyces cerevisiae ADH1 promoter" gene 1713..3359 /gene="amdS" CDS 1713..3359 /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 4067..4647 /note="modified Kluyveromyces lactis LAC4 promoter 5' end" gene complement(5861..6721) /gene="bla" CDS complement(5861..6721) /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 7475..9107 /note="modified Kluyveromyces lactis LAC4 promoter 3' end" ORIGIN 1 aagcttgaaa aaaatgaaat tctctactat attagccgca tctactgctt taatttccgt 61 tgttatggct gctccagttt ctaccgaaac tgacatcgac gatcttccaa tatcggttcc 121 agaagaagcc ttgattggat tcattgactt aaccggggat gaagtttcct tgttgcctgt 181 taataacgga acccacactg gtattctatt cttaaacacc accatcgctg aagctgcttt 241 cgctgacaag gatgatctcg agaaaagaga ggctgaagct agaagagctc atatgtccat 301 gggcggccgc gatatcgtcg acggatccga attccctgca ggtaattaaa taaaggcctt 361 gaatcgagaa tttatactta gataagtatg tacttacagg tatatttcta tgagatactg 421 atgtatacat gcatgataat atttaaacgg ttattagtgc cgattgtctt gtgcgataat 481 gacgttccta tcaaagcaat acacttacca cctattacat gggccaagaa aatattttcg 541 aacttgttta gaatattagc acagagtata tgatgttatc cgttagatta tgcatgattc 601 attcctacaa ctttttcgta gcataaggat taattacttg gatgccaata aaaaaaaaaa 661 acatcgagaa aatttcagca tgctcagaaa caattgcagt gtatcaaagt aaaaaaaaga 721 ttttcactac atgttccttt tgaagaaaga aaatcatgga acattagatt tacaaaaatt 781 taaccaccgc tgattaacga ttagaccgtt aagcgcacaa caggttatta gtacagagaa 841 agcattctgt ggtgttgccc cggactttct tttgcgacat aggtaaatcg aataccatca 901 tactatcttt tccaatgact ccctaaagaa agactcttct tcgatgttgt atacgttgga 961 gcatagggca agaattgtgg cttgagatca tccttttgtt gtttccgggt gtacaatatg 1021 gacttcctct tttctggcaa ccaaacccat acatcgggat tcctataata ccttcgttgg 1081 tctccctaac atgtaggtgg cggaggggag atatacaata gaacagatac cagacaagac 1141 ataatgggct aaacaagact acaccaatta cactgcctca ttgatggtgg tacataacga 1201 actaatactg tagccctaga cttgatagcc atcatcatat cgaagtttca ctaccctttt 1261 tccatttgcc atctattgaa gtaataatag gcgcatgcaa cttcttttct ttttttttct 1321 tttctctctc ccccgttgtt gtctcaccat atccgcaatg acaaaaaaat gatggaagac 1381 actaaaggaa aaaattaacg acaaagacag caccaacaga tgtcgttgtt ccagagctga 1441 tgaggggtat ctcgaagcac acgaaacttt ttccttcctt cattcacgca cactactctc 1501 taatgagcaa cggtatacgg ccttccttcc agttacttga atttgaaata aaaaaaagtt 1561 tgctgtcttg ctatcaagta taaatagacc tgcaattatt aatcttttgt ttcctcgtca 1621 ttgttctcgt tccctttctt ccttgtttct ttttctgcac aatatttcaa gctataccaa 1681 gcatacaatc aagcaattcc agatctgcca ccatgcctca atcctgggaa gaactggccg 1741 ctgataagcg cgcccgcctc gcaaaaacca tccctgatga atggaaagtc cagacgctgc 1801 ctgcggaaga cagcgttatt gatttcccaa agaaatcggg catcctttca gaggccgaac 1861 tgaagatcac agaggcttcc gctgcggatc ttgtgtccaa gctggcggcc ggagagttga 1921 cctcggtgga agttacgcta gcattctgta aacgggcagc aatcgcccag cagttaacaa 1981 actgcgccca cgagttcttc cctgacgccg ctctcgcgca ggcaagggaa ctcgatgaat 2041 actacgcaaa gcacaagaga cccgttggtc cacttcatgg cctccccatc tctctcaaag 2101 accagcttcg agtcaagggc tacgaaacat caatgggcta catctcatgg ctaaacaagt 2161 acgacgaagg ggactcggtt ctgacaacca tgctccgcaa agccggtgcc gtcttctacg 2221 tcaagacctc tgtcccgcag accctgatgg tctgcgagac agtcaacaac atcatcgggc 2281 gcaccgtcaa cccacgcaac aagaactggt cgtgcggcgg cagttctggt ggtgagggtg 2341 cgatcgttgg gattcgtggt ggcgtcatcg gtgtaggaac ggacatcggt ggctcgattc 2401 gagtgccggc cgcgttcaac ttcctgtacg gtctaaggcc gagtcatggg cggctgccgt 2461 atgcaaagat ggcgaacagc atggagggtc aggagacggt gcacagcgtt gtcgggccga 2521 ttacgcactc tgttgaggac ctccgcctct tcaccaaatc cgtcctcggt caggagcctt 2581 ggaaatacga ctccaaggtc atccccatgc cctggcgcca gtccgagtcg gacattattg 2641 cctccaagat caagaacggc gggctcaata tcggctacta caacttcgac ggcaatgtcc 2701 ttccacaccc tcctatcctg cgcggcgtgg aaactaccgt cgccgcactc gccaaagccg 2761 gtcacaccgt gaccccgtgg acgccataca agcacgattt cggccacgat ctcatctccc 2821 atatctacgc ggctgacggc agcgccgacg taatgcgcga catcagtgca tccggcgagc 2881 cggcgattcc aaatatcaaa gacctactga acccgaacat caaagctgtt aacatgaacg 2941 agctctggga cacgcatctc cagaagtgga attaccagat ggagtacctt gagaaatggc 3001 gggaggctga agaaaaggcc gggaaggaac tggacgccat catcgcgccg attacgccta 3061 ccgctgcggt acggcatgac cagttccggt actatgggta tgcctctgtg atcaacctgc 3121 tggatttcac gagcgtggtt gttccggtta cctttgcgga taagaacatc gataagaaga 3181 atgagagttt caaggcggtt agtgagcttg atgccctcgt gcaggaagag tatgatccgg 3241 aggcgtatca tggggcaccg gttgcagtgc aggttatcgg acggagactc agtgaagaga 3301 ggacgttggc gattgcagag gaagtgggga agttgctggg aaatgtggtg actccatagc 3361 ccgggggggg ctcgatcccc tcgcgagttg gttcagctgc tgcctgaggc tggacgacct 3421 cgcggagttc taccggcagt gcaaatccgt cggcatccag gaaaccagca gcggctatcc 3481 gcgcatccat gcccccgaac tgcaggagtg gggaggcacg atggccgctt tggtcgatct 3541 agattacgtg gaagaaaggt agtaaaagta gtagtataag tagtaaaaag aggtaaaaag 3601 agaaaaccgg ctacatacta gagaagcacg tacacaaaaa ctcataggca cttcatcata 3661 cgacagtttc ttgatgcatt ataatagtgt attagatatt ttcagaaata tgcatagaac 3721 ctcctcttgc ctttactttt tatacataga acattggcag atttacttac actactttgt 3781 ttctacgcca tttcttttgt tttcaacact tagacaagtt gttgagaacc ggactactaa 3841 aaagcaatgt tcccactgaa aatcatgtac ctgcagcata ataaccccct aattctgcat 3901 cgatccagta tgtttttttt tctctactca tttttacctg aagatagagc ttctaaaaca 3961 aaaaaaatca gtgattacat gcatattgtg tgttctagta accaaaggaa aggaacagat 4021 agataaaatt ccgagactgt caaattaggt ttttttcttt ttttttggcg ggagtcagtg 4081 ggccgaaata tgttcttggc ctagaactta atctggtttg atcatgccaa tacttgcctg 4141 agtgcccgac tttttgccca ccctcttgcc ttctgtcatc cttcaaaacc cacctgtttt 4201 ccagccgtat cttcgctcgc atctacacat actgtgccat atcttgtgtg tagccggacg 4261 tgactatgac caaaaacaaa caaggagaac tgttcgccga tttgtaacac tcctgcatcc 4321 atccaagtgg gtatgcgcta tgcaatgtta agctaggtca ggtcagacca ggtccaagga 4381 cagcaacttg actgtatgca acctttacca tctttgcaca gaacatactt gtagctagct 4441 agttacactt atggaccgaa aaggcacccc accatgtctg tccggcttta gagtacggcc 4501 gcagaccgct gatttgcctt gccaagcagt agtcacaatg catcgcatga gcacacgggc 4561 acgggcacgg gcacaggaac cattggcaaa aataccagat acactatacc gacgtatatc 4621 aagcccaagt ttaaaattcc taaatttccg cggctacttt tcaattccct atagtgagtc 4681 gtattaaatt cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 4741 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 4801 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 4861 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 4921 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 4981 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 5041 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 5101 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 5161 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 5221 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 5281 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 5341 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 5401 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 5461 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 5521 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 5581 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 5641 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 5701 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 5761 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 5821 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 5881 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 5941 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 6001 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 6061 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 6121 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 6181 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 6241 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 6301 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 6361 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 6421 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 6481 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 6541 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 6601 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 6661 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 6721 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 6781 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 6841 aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc 6901 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt 6961 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag 7021 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt 7081 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt 7141 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt 7201 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt 7261 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc attcgccatt 7321 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct 7381 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc 7441 acgacgttgt aaaacgacgg ccagtgccaa gctcccgcgg ggatcgactc ataaaatagt 7501 aaccttctaa tgcgtatcta ttgactacca accattagtg tggttgcaga aggcggaatt 7561 ttcccttctt cgaatttagc ttgctttttc attttttatt ttccattttt cagtttttgt 7621 ttgtgtcgaa tttagccagt tgcttctcca agatgaaaaa aacccctgcg cagtttctgt 7681 gctgcaagat cctaatcgac ttttccaccc cccacaaaag taaatgttct tttgttacat 7741 tcgcgtgggt agctagctcc ccgaatcttc aaaggactta gggactgcac tacatcagag 7801 tgtgttcacc tggtttgctg cctggtttga aagaaaagag cagggaactc gcgggttccc 7861 ggcgaataat catgcgatag tcctttggcc ttccaagtcg catgtagagt agacaacaga 7921 cagggagggc aggaaggatc tttcactgag atcctgtatc ttgttgggta agtcggatga 7981 aaggggaatc gtatgagatt ggagaggatg cggaagaggt aacgcctttt gttaacttgt 8041 ttaattatta tggggcaggc gagaggggga ggaatgtatg tgtgtgaggc gggcgagacg 8101 gagccatcca ggccaggtag aaatagagaa agccgaatgt tagacaatat ggcagcgtag 8161 tagagtaggt aggtaggcaa gtactgctag caaagaggag aagggtaagc tcactcttcg 8221 cattccacac cgttagtgtg tcagtttgaa caaaaaaaca atcatcatac caattgatgg 8281 actgtggact ggcttttgga acggcttttc ggactgcgat tattcgtgag gaatcaaggt 8341 aggaatttgg tcatatttac ggacaacagt gggtgattcc catatcgagt aggaaaacga 8401 gatcatggta tcctcagata tgttgcggaa atctgttcac cgcaaagttc agggtgctct 8461 ggtgggtttc ggttggtctt tgctttgctt ctcccttgtc ttgcatgtta ataatagcct 8521 agcctgtgag ccgaaactta gggtaggctt agtgttggaa cgtacatatc tatcacgttg 8581 acttggttta accaggcgac ctggtagcca gccataccca cacacgtttt ttgtatcttc 8641 agtatagttg tgaaaagtgt agcggaaatt tgtggtccga gcaacagcgt ctttttctag 8701 tagtgcggtc ggttacttgg ttgacattgg tatttggact ttgttgctac accattcact 8761 acttgaagtc gagtgtgaag ggtatgattt ctagtggtga acacctttag ttacgtaatg 8821 ttttcattgc tgttttactt gagatttcga ttgagaaaaa ggtatttaat agctcgaatc 8881 aatgtgagaa cagagagaag atgttcttcc ctaactcgaa aggtatatga ggcttgtgtt 8941 tcttaggaga attattattc ttttgttatg ttgcgcttgt agttggaaaa ggtgaagaga 9001 caaaagctgg aattgtgagc ggataacaag ctcaacactt gaaatttagg aaagagcaga 9061 atttggcaaa aaaaataaaa aaaaaataaa cacacatact catcgag //