LOCUS FJ010198 9589 bp DNA circular SYN 12-AUG-2008 DEFINITION Expression vector pKLAC1-Gluc, complete sequence. ACCESSION FJ010198 VERSION FJ010198.1 KEYWORDS . SOURCE Expression vector pKLAC1-Gluc ORGANISM Expression vector pKLAC1-Gluc other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 9589) AUTHORS Colussi,P.A., Ganatra,M.B., Read,J.D. and Taron,C.H. TITLE A vector for expression of Gaussia princeps luciferase tagged proteins in the yeast Kluyveromyces lactis JOURNAL Unpublished REFERENCE 2 (bases 1 to 9589) AUTHORS Colussi,P.A., Ganatra,M.B., Read,J.D. and Taron,C.H. TITLE Direct Submission JOURNAL Submitted (12-AUG-2008) Gene Expression Division, New England Biolabs, 240 County Road, Ipswich, MA 01938 COMMENT Bankit Comment: HAS CLONING VECTOR. FEATURES Location/Qualifiers source 1..9589 /organism="Kluyveromyces lactis" /mol_type="other DNA" /db_xref="taxon:28985" misc_feature 14..331 /note="Kluyveromyces lactis alpha mating factor pre-pro secretion leader" misc_feature 290..331 /note="multiple cloning site" gene <332..838 /gene="Gluc" CDS <332..838 /gene="Gluc" /note="luciferase" /codon_start=1 /product="Gaussia princeps luciferase" /translation="KPTENNEDFNIVAVASNFATTDLDADRGKLPGKKLPLEVLKEME ANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESAQGGIGEAIVDIPEIP GFKDLEPMEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQRCATFASKIQGQVD KIKGAGGD" terminator 853..1435 /note="Kluyveromyces lactis LAC4 transcription terminator" promoter 1492..2194 /note="Saccharomyces cerevisiae ADH1 promoter" gene 2195..3841 /gene="amdS" CDS 2195..3841 /gene="amdS" /note="acetamidase; confers ability to process acetamide for selection in Kluyveromyces lactis" /codon_start=1 /product="Aspergillus nidulans acetamidase" /translation="MPQSWEELAADKRARLAKTIPDEWKVQTLPAEDSVIDFPKKSGI LSEAELKITEASAADLVSKLAAGELTSVEVTLAFCKRAAIAQQLTNCAHEFFPDAALA QARELDEYYAKHKRPVGPLHGLPISLKDQLRVKGYETSMGYISWLNKYDEGDSVLTTM LRKAGAVFYVKTSVPQTLMVCETVNNIIGRTVNPRNKNWSCGGSSGGEGAIVGIRGGV IGVGTDIGGSIRVPAAFNFLYGLRPSHGRLPYAKMANSMEGQETVHSVVGPITHSVED LRLFTKSVLGQEPWKYDSKVIPMPWRQSESDIIASKIKNGGLNIGYYNFDGNVLPHPP ILRGVETTVAALAKAGHTVTPWTPYKHDFGHDLISHIYAADGSADVMRDISASGEPAI PNIKDLLNPNIKAVNMNELWDTHLQKWNYQMEYLEKWREAEEKAGKELDAIIAPITPT AAVRHDQFRYYGYASVINLLDFTSVVVPVTFADKNIDKKNESFKAVSELDALVQEEYD PEAYHGAPVAVQVIGRRLSEERTLAIAEEVGKLLGNVVTP" promoter 4550..5130 /note="modified Kluyveromyces lactis LAC4 promoter 5' end" gene complement(6343..7203) /gene="bla" CDS complement(6343..7203) /gene="bla" /note="confers ampicillin resistance in Escherichia coli" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" promoter 7957..9589 /note="modified Kluyveromyces lactis LAC4 promoter 3' end" BASE COUNT 2527 a 2274 c 2292 g 2496 t 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 agaagagcta gatctcctag 301 gggtaccgtc gacggcgcgc ctgcggccgc taagcccacc gagaacaacg aagacttcaa 361 catcgtggcc gtggccagca acttcgcgac cacggatctc gatgctgatc gcgggaagtt 421 gcccggcaag aagctgccgc tggaggtgct caaagagatg gaagccaatg cccggaaagc 481 tggctgcacc aggggctgtc tgatctgcct gtcccacatc aagtgcacgc ccaagatgaa 541 gaagttcatc ccaggacgct gccacaccta cgaaggcgac aaagagtccg cacagggcgg 601 cataggcgag gcgatcgtcg acattcctga gattcctggg ttcaaggact tggagcccat 661 ggagcagttc atcgcacagg tcgatctgtg tgtggactgc acaactggct gcctcaaagg 721 gcttgccaac gtgcagtgtt ctgacctgct caagaagtgg ctgccgcaac gctgtgcgac 781 ctttgccagc aagatccagg gccaggtgga caagatcaag ggggccggtg gtgactaacc 841 ttgaatcgag aatttatact tagataagta tgtacttaca ggtatatttc tatgagatac 901 tgatgtatac atgcatgata atatttaaac ggttattagt gccgattgtc ttgtgcgata 961 atgacgttcc tatcaaagca atacacttac cacctattac atgggccaag aaaatatttt 1021 cgaacttgtt tagaatatta gcacagagta tatgatgata tccgttagat tatgcatgat 1081 tcattcctac aactttttcg tagcataagg attaattact tggatgccaa taaaaaaaaa 1141 aaacatcgag aaaatttcag catgctcaga aacaattgca gtgtatcaaa gtaaaaaaaa 1201 gattttcact acatgttcct tttgaagaaa gaaaatcatg gaacattaga tttacaaaaa 1261 tttaaccacc gctgattaac gattagaccg ttaagcgcac aacaggttat tagtacagag 1321 aaagcattct gtggtgttgc cccggacttt cttttgcgac ataggtaaat cgaataccat 1381 catactatct tttccaatga ctccctaaag aaagactctt cttcgatgtt gtatacgttg 1441 gagcataggg caagaattgt ggcttgagat catccttttg ttgtttccgg gtgtacaata 1501 tggacttcct cttttctggc aaccaaaccc atacatcggg attcctataa taccttcgtt 1561 ggtctcccta acatgtaggt ggcggagggg agatatacaa tagaacagat accagacaag 1621 acataatggg ctaaacaaga ctacaccaat tacactgcct cattgatggt ggtacataac 1681 gaactaatac tgtagcccta gacttgatag ccatcatcat atcgaagttt cactaccctt 1741 tttccatttg ccatctattg aagtaataat aggcgcatgc aacttctttt cttttttttt 1801 cttttctctc tcccccgttg ttgtctcacc atatccgcaa tgacaaaaaa atgatggaag 1861 acactaaagg aaaaaattaa cgacaaagac agcaccaaca gatgtcgttg ttccagagct 1921 gatgaggggt atctcgaagc acacgaaact ttttccttcc ttcattcacg cacactactc 1981 tctaatgagc aacggtatac ggccttcctt ccagttactt gaatttgaaa taaaaaaaag 2041 tttgctgtct tgctatcaag tataaataga cctgcaatta ttaatctttt gtttcctcgt 2101 cattgttctc gttccctttc ttccttgttt ctttttctgc acaatatttc aagctatacc 2161 aagcatacaa tcaaggaatt ccggatccgc caccatgcct caatcctggg aagaactggc 2221 cgctgataag cgcgcccgcc tcgcaaaaac catccctgat gaatggaaag tccagacgct 2281 gcctgcggaa gacagcgtta ttgatttccc aaagaaatcg gggatccttt cagaggccga 2341 actgaagatc acagaggctt ccgctgcgga tcttgtgtcc aagctggcgg ccggagagtt 2401 gacctcggtg gaagttacgc tagcattctg taaacgggca gcaatcgccc agcagttaac 2461 aaactgcgcc cacgagttct tccctgacgc cgctctcgcg caggcaaggg aactcgatga 2521 atactacgca aagcacaaga gacccgttgg tccactccat ggcctcccca tctctctcaa 2581 agaccagctt cgagtcaagg gctacgaaac atcaatgggc tacatctcat ggctaaacaa 2641 gtacgacgaa ggggactcgg ttctgacaac catgctccgc aaagccggtg ccgtcttcta 2701 cgtcaagacc tctgtcccgc agaccctgat ggtctgcgag acagtcaaca acatcatcgg 2761 gcgcaccgtc aacccacgca acaagaactg gtcgtgcggc ggcagttctg gtggtgaggg 2821 tgcgatcgtt gggattcgtg gtggcgtcat cggtgtagga acggatatcg gtggctcgat 2881 tcgagtgccg gccgcgttca acttcctgta cggtctaagg ccgagtcatg ggcggctgcc 2941 gtatgcaaag atggcgaaca gcatggaggg tcaggagacg gtgcacagcg ttgtcgggcc 3001 gattacgcac tctgttgagg acctccgcct cttcaccaaa tccgtcctcg gtcaggagcc 3061 atggaaatac gactccaagg tcatccccat gccctggcgc cagtccgagt cggacattat 3121 tgcctccaag atcaagaacg gcgggctcaa tatcggctac tacaacttcg acggcaatgt 3181 ccttccacac cctcctatcc tgcgcggcgt ggaaactacc gtcgccgcac tcgccaaagc 3241 cggtcacacc gtgaccccgt ggacgccata caagcacgat ttcggccacg atctcatctc 3301 ccatatctac gcggctgacg gcagcgccga cgtaatgcgc gatatcagtg catccggcga 3361 gccggcgatt ccaaatatca aagacctact gaacccgaac atcaaagctg ttaacatgaa 3421 cgagctctgg gacacgcatc tccagaagtg gaattaccag atggagtacc ttgagaaatg 3481 gcgggaggct gaagaaaagg ccgggaagga actggacgcc atcatcgcgc cgattacgcc 3541 taccgctgcg gtacggcatg accagttccg gtactatggg tatgcctctg tgatcaacct 3601 gctggatttc acgagcgtgg ttgttccggt tacctttgcg gataagaaca tcgataagaa 3661 gaatgagagt ttcaaggcgg ttagtgagct tgatgccctc gtgcaggaag agtatgatcc 3721 ggaggcgtac catggggcac cggttgcagt gcaggttatc ggacggagac tcagtgaaga 3781 gaggacgttg gcgattgcag aggaagtggg gaagttgctg ggaaatgtgg tgactccata 3841 gcccgggggg ggctcgatcc cctcgcgagt tggttcagct gctgcctgag gctggacgac 3901 ctcgcggagt tctaccggca gtgcaaatcc gtcggcatcc aggaaaccag cagcggctat 3961 ccgcgcatcc atgcccccga actgcaggag tggggaggca cgatggccgc tttggtcgat 4021 ctagattacg tggaagaaag gtagtaaaag tagtagtata agtagtaaaa agaggtaaaa 4081 agagaaaacc ggctacatac tagagaagca cgtacacaaa aactcatagg cacttcatca 4141 tacgacagtt tcttgatgca ttataatagt gtattagata ttttcagaaa tatgcataga 4201 acctcctctt gcctttactt tttatacata gaacattggc agatttactt acactacttt 4261 gtttctacgc catttctttt gttttcaaca cttagacaag ttgttgagaa ccggactact 4321 aaaaagcaat gttcccactg aaaatcatgt acctgcagga taataacccc ctaattctgc 4381 atcgatccag tatgtttttt tttctctact catttttacc tgaagataga gcttctaaaa 4441 caaaaaaaat cagtgattac atgcatattg tgtgttctag taaccaaagg aaaggaacag 4501 atagataaaa ttccgagact gtcaaattag gtttttttct ttttttttgg cgggagtcag 4561 tgggccgaaa tatgttcttg gcctagaact taatctggtt tgatcatgcc aatacttgcc 4621 tgagtgcccg actttttgcc caccctcttg ccttctgtca tccttcaaaa cccacctgtt 4681 ttccagccgt atcttcgctc gcatctacac atactgtgcc atatcttgtg tgtagccgga 4741 cgtgactatg accaaaaaca aacaaggaga actgttcgcc gatttgtaac actcctgcat 4801 ccatccaagt gggtatgcgc tatgcaatgt taagctaggt caggtcagac caggtccaag 4861 gacagcaact tgactgtatg caacctttac catctttgca cagaacatac ttgtagctag 4921 ctagttacac ttatggaccg aaaaggcacc ccaccatgtc tgtccggctt tagagtacgg 4981 ccgcagaccg ctgatttgcc ttgccaagca gtagtcacaa tgcatcgcat gagcacacgg 5041 gcacgggcac gggcacagga accattggca aaaataccag atacactata ccgacgtata 5101 tcaagcccaa gtttaaaatt cctaaatttc cgcggctact tttcaattcc ctatagtgag 5161 tcgtattaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac 5221 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt 5281 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc 5341 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg 5401 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt 5461 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa 5521 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc 5581 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag 5641 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt 5701 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg 5761 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg 5821 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg 5881 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac 5941 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg 6001 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt 6061 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg 6121 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc 6181 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt 6241 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt 6301 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag 6361 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt 6421 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc 6481 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc 6541 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg 6601 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac 6661 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg 6721 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc 6781 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact 6841 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc 6901 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat 6961 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc 7021 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac 7081 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa 7141 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact 7201 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg 7261 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg 7321 aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac 7381 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc 7441 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt 7501 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg 7561 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac 7621 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta 7681 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat 7741 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt ccattcgcca 7801 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag 7861 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag 7921 tcacgacgtt gtaaaacgac ggccagtgcc aagctcccgc ggggatcgac tcataaaata 7981 gtaaccttct aatgcgtatc tattgactac caaccattag tgtggttgca gaaggcggaa 8041 ttctcccttc ttcgaattca gcttgctttt tcatttttta ttttccattt ttcagttttt 8101 gtttgtgtcg aatttagcca gttgcttctc caagatgaaa aaaacccctg cgcagtttct 8161 gtgctgcaag atcctaatcg acttttccac cccccacaaa agtaaatgtt cttttgttac 8221 attcgcgtgg gtagctagct ccccgaatct tcaaaggact tagggactgc actacatcag 8281 agtgtgttca cctggtttgc tgcctggttt gaaagaaaag agcagggaac tcgcgggttc 8341 ccggcgaata atcatgcgat agtcctttgg ccttccaagt cgcatgtaga gtagacaaca 8401 gacagggagg gcaggaagga tctttcactg agatcctgta tcttgttggg taagtcggat 8461 gaaaggggaa tcgtatgaga ttggagagga tgcggaagag gtaacgcctt ttgttaactt 8521 gtttaattat tatggggcag gcgagagggg gaggaatgta tgtgtgtgag gcgggcgaga 8581 cggagccatc caggccaggt agaaatagag aaagccgaat gttagacaat atggcagcgt 8641 agtagagtag gtaggtaggc aagtactgct agcaaagagg agaagggtaa gctcactctt 8701 cgcattccac accgttagtg tgtcagtttg aacaaaaaaa caatcatcat accaattgat 8761 ggactgtgga ctggcttttg gaacggcttt tcggactgcg attattcgtg aggaatcaag 8821 gtaggaattt ggtcatattt acggacaaca gtgggtgatt cccatatgga gtaggaaaac 8881 gagatcatgg tatcctcaga tatgttgcgg aattctgttc accgcaaagt tcagggtgct 8941 ctggtgggtt tcggttggtc tttgctttgc ttctcccttg tcttgcatgt taataatagc 9001 ctagcctgtg agccgaaact tagggtaggc ttagtgttgg aacgtacata tgtatcacgt 9061 tgacttggtt taaccaggcg acctggtagc cagccatacc cacacacgtt ttttgtatct 9121 tcagtatagt tgtgaaaagt gtagcggaaa tttgtggtcc gagcaacagc gtctttttct 9181 agtagtgcgg tcggttactt ggttgacatt ggtatttgga ctttgttgct acaccattca 9241 ctacttgaag tcgagtgtga agggtatgat ttctagtggt gaacaccttt agttacgtaa 9301 tgttttcatt gctgttttac ttgagatttc gattgagaaa aaggtattta atagctcgaa 9361 tcaatgtgag aacagagaga agatgttctt ccctaactcg aaaggtatat gaggcttgtg 9421 tttcttagga gaattattat tcttttgtta tgttgcgctt gtagttggaa aaggtgaaga 9481 gacaaaagct ggaattgtga gcggataaca agctcaacac ttgaaattta ggaaagagca 9541 gaatttggca aaaaaaataa aaaaaaaata aacacacata ctcatcgag //