LOCUS pKLAC1 9091 bp DNA circular SYN 18-OCT-2007 DEFINITION Expression vector pKLAC1, complete sequence. ACCESSION VERSION KEYWORDS . SOURCE Expression vector pKLAC1 ORGANISM Expression vector pKLAC1 other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 9091) AUTHORS Colussi,P.A. and Taron,C.H. TITLE Kluyveromyces lactis LAC4 promoter variants that lack function in bacteria but retain full function in K. lactis JOURNAL Appl. Environ. Microbiol. 71 (11), 7092-7098 (2005) PUBMED 16269745 REFERENCE 2 (bases 1 to 9091) AUTHORS Colussi,P.A. and Taron,C.H. TITLE Direct Submission JOURNAL Submitted (18-OCT-2007) Research Department, New England Biolabs, 240 County Road, Ipswich, MA 01938, USA COMMENT See also GenBank accession AY968582. FEATURES Location/Qualifiers source 1..9091 /organism="Expression vector pKLAC1" /mol_type="other DNA" gene 14..334 /gene="alpha-MF" CDS 14..334 /gene="alpha-MF" /codon_start=1 /product="Kluyveromyces lactis alpha mating factor pre-pro secretion leader" /translation="MKFSTILAASTALISVVMAAPVSTETDIDDLPISVPEEALIGFI DLTGDEVSLLPVNNGTHTGILFLNTTIAEAAFADKDDLEKREAEARRARSPRGTVDGA PAAA" misc_feature 257..343 /note="multiple cloning site (XhoI-StuI)" terminator 355..937 /note="Kluyveromyces lactis LAC4 transcription terminator; stops transcription from the alpha-MF/target gene fusion" promoter 994..1696 /note="Saccharomyces cerevisiae ADH1 promoter; drives amdS transcription" gene 1697..3343 /gene="amdS" CDS 1697..3343 /gene="amdS" /note="selectable marker allowing growth on acetamide as a sole nitrogen source" /codon_start=1 /product="acetamidase" /translation="MPQSWEELAADKRARLAKTIPDEWKVQTLPAEDSVIDFPKKSGI LSEAELKITEASAADLVSKLAAGELTSVEVTLAFCKRAAIAQQLTNCAHEFFPDAALA QARELDEYYAKHKRPVGPLHGLPISLKDQLRVKGYETSMGYISWLNKYDEGDSVLTTM LRKAGAVFYVKTSVPQTLMVCETVNNIIGRTVNPRNKNWSCGGSSGGEGAIVGIRGGV IGVGTDIGGSIRVPAAFNFLYGLRPSHGRLPYAKMANSMEGQETVHSVVGPITHSVED LRLFTKSVLGQEPWKYDSKVIPMPWRQSESDIIASKIKNGGLNIGYYNFDGNVLPHPP ILRGVETTVAALAKAGHTVTPWTPYKHDFGHDLISHIYAADGSADVMRDISASGEPAI PNIKDLLNPNIKAVNMNELWDTHLQKWNYQMEYLEKWREAEEKAGKELDAIIAPITPT AAVRHDQFRYYGYASVINLLDFTSVVVPVTFADKNIDKKNESFKAVSELDALVQEEYD PEAYHGAPVAVQVIGRRLSEERTLAIAEEVGKLLGNVVTP" promoter 4052..4632 /note="modified Kluyveromyces lactis LAC4 promoter 5' end" misc_feature 4303..4314 /note="BstXI restriction site (linearization, alternate)" misc_feature 4632..4637 /note="SacII restriction site (linearization)" rep_origin 5086..5674 /note="pMB1 origin of replication (counterclockwise) (RNAII -35 to RNA/DNA switch point)" gene complement(5845..6705) /gene="bla" CDS complement(5845..6705) /gene="bla" /note="ampR (confers resistance to ampicillin)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" promoter 7459..9091 /note="modified Kluyveromyces lactis LAC4 promoter 3' end" misc_feature 7459..7464 /note="SacII restriction site (linearization)" misc_feature 7516..7527 /note="BstXI restriction site (linearization, alternate)" BASE COUNT 2408 a 2132 c 2139 g 2412 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 ttaattaagg ccttgaatcg agaatttata 361 cttagataag tatgtactta caggtatatt tctatgagat actgatgtat acatgcatga 421 taatatttaa acggttatta gtgccgattg tcttgtgcga taatgacgtt cctatcaaag 481 caatacactt accacctatt acatgggcca agaaaatatt ttcgaacttg tttagaatat 541 tagcacagag tatatgatga tatccgttag attatgcatg attcattcct acaacttttt 601 cgtagcataa ggattaatta cttggatgcc aataaaaaaa aaaaacatcg agaaaatttc 661 agcatgctca gaaacaattg cagtgtatca aagtaaaaaa aagattttca ctacatgttc 721 cttttgaaga aagaaaatca tggaacatta gatttacaaa aatttaacca ccgctgatta 781 acgattagac cgttaagcgc acaacaggtt attagtacag agaaagcatt ctgtggtgtt 841 gccccggact ttcttttgcg acataggtaa atcgaatacc atcatactat cttttccaat 901 gactccctaa agaaagactc ttcttcgatg ttgtatacgt tggagcatag ggcaagaatt 961 gtggcttgag atcatccttt tgttgtttcc gggtgtacaa tatggacttc ctcttttctg 1021 gcaaccaaac ccatacatcg ggattcctat aataccttcg ttggtctccc taacatgtag 1081 gtggcggagg ggagatatac aatagaacag ataccagaca agacataatg ggctaaacaa 1141 gactacacca attacactgc ctcattgatg gtggtacata acgaactaat actgtagccc 1201 tagacttgat agccatcatc atatcgaagt ttcactaccc tttttccatt tgccatctat 1261 tgaagtaata ataggcgcat gcaacttctt ttcttttttt ttcttttctc tctcccccgt 1321 tgttgtctca ccatatccgc aatgacaaaa aaatgatgga agacactaaa ggaaaaaatt 1381 aacgacaaag acagcaccaa cagatgtcgt tgttccagag ctgatgaggg gtatctcgaa 1441 gcacacgaaa ctttttcctt ccttcattca cgcacactac tctctaatga gcaacggtat 1501 acggccttcc ttccagttac ttgaatttga aataaaaaaa agtttgctgt cttgctatca 1561 agtataaata gacctgcaat tattaatctt ttgtttcctc gtcattgttc tcgttccctt 1621 tcttccttgt ttctttttct gcacaatatt tcaagctata ccaagcatac aatcaaggaa 1681 ttccggatcc gccaccatgc ctcaatcctg ggaagaactg gccgctgata agcgcgcccg 1741 cctcgcaaaa accatccctg atgaatggaa agtccagacg ctgcctgcgg aagacagcgt 1801 tattgatttc ccaaagaaat cggggatcct ttcagaggcc gaactgaaga tcacagaggc 1861 ttccgctgcg gatcttgtgt ccaagctggc ggccggagag ttgacctcgg tggaagttac 1921 gctagcattc tgtaaacggg cagcaatcgc ccagcagtta acaaactgcg cccacgagtt 1981 cttccctgac gccgctctcg cgcaggcaag ggaactcgat gaatactacg caaagcacaa 2041 gagacccgtt ggtccactcc atggcctccc catctctctc aaagaccagc ttcgagtcaa 2101 gggctacgaa acatcaatgg gctacatctc atggctaaac aagtacgacg aaggggactc 2161 ggttctgaca accatgctcc gcaaagccgg tgccgtcttc tacgtcaaga cctctgtccc 2221 gcagaccctg atggtctgcg agacagtcaa caacatcatc gggcgcaccg tcaacccacg 2281 caacaagaac tggtcgtgcg gcggcagttc tggtggtgag ggtgcgatcg ttgggattcg 2341 tggtggcgtc atcggtgtag gaacggatat cggtggctcg attcgagtgc cggccgcgtt 2401 caacttcctg tacggtctaa ggccgagtca tgggcggctg ccgtatgcaa agatggcgaa 2461 cagcatggag ggtcaggaga cggtgcacag cgttgtcggg ccgattacgc actctgttga 2521 ggacctccgc ctcttcacca aatccgtcct cggtcaggag ccatggaaat acgactccaa 2581 ggtcatcccc atgccctggc gccagtccga gtcggacatt attgcctcca agatcaagaa 2641 cggcgggctc aatatcggct actacaactt cgacggcaat gtccttccac accctcctat 2701 cctgcgcggc gtggaaacta ccgtcgccgc actcgccaaa gccggtcaca ccgtgacccc 2761 gtggacgcca tacaagcacg atttcggcca cgatctcatc tcccatatct acgcggctga 2821 cggcagcgcc gacgtaatgc gcgatatcag tgcatccggc gagccggcga ttccaaatat 2881 caaagaccta ctgaacccga acatcaaagc tgttaacatg aacgagctct gggacacgca 2941 tctccagaag tggaattacc agatggagta ccttgagaaa tggcgggagg ctgaagaaaa 3001 ggccgggaag gaactggacg ccatcatcgc gccgattacg cctaccgctg cggtacggca 3061 tgaccagttc cggtactatg ggtatgcctc tgtgatcaac ctgctggatt tcacgagcgt 3121 ggttgttccg gttacctttg cggataagaa catcgataag aagaatgaga gtttcaaggc 3181 ggttagtgag cttgatgccc tcgtgcagga agagtatgat ccggaggcgt accatggggc 3241 accggttgca gtgcaggtta tcggacggag actcagtgaa gagaggacgt tggcgattgc 3301 agaggaagtg gggaagttgc tgggaaatgt ggtgactcca tagcccgggg ggggctcgat 3361 cccctcgcga gttggttcag ctgctgcctg aggctggacg acctcgcgga gttctaccgg 3421 cagtgcaaat ccgtcggcat ccaggaaacc agcagcggct atccgcgcat ccatgccccc 3481 gaactgcagg agtggggagg cacgatggcc gctttggtcg atctagatta cgtggaagaa 3541 aggtagtaaa agtagtagta taagtagtaa aaagaggtaa aaagagaaaa ccggctacat 3601 actagagaag cacgtacaca aaaactcata ggcacttcat catacgacag tttcttgatg 3661 cattataata gtgtattaga tattttcaga aatatgcata gaacctcctc ttgcctttac 3721 tttttataca tagaacattg gcagatttac ttacactact ttgtttctac gccatttctt 3781 ttgttttcaa cacttagaca agttgttgag aaccggacta ctaaaaagca atgttcccac 3841 tgaaaatcat gtacctgcag gataataacc ccctaattct gcatcgatcc agtatgtttt 3901 tttttctcta ctcattttta cctgaagata gagcttctaa aacaaaaaaa atcagtgatt 3961 acatgcatat tgtgtgttct agtaaccaaa ggaaaggaac agatagataa aattccgaga 4021 ctgtcaaatt aggttttttt cttttttttt ggcgggagtc agtgggccga aatatgttct 4081 tggcctagaa cttaatctgg tttgatcatg ccaatacttg cctgagtgcc cgactttttg 4141 cccaccctct tgccttctgt catccttcaa aacccacctg ttttccagcc gtatcttcgc 4201 tcgcatctac acatactgtg ccatatcttg tgtgtagccg gacgtgacta tgaccaaaaa 4261 caaacaagga gaactgttcg ccgatttgta acactcctgc atccatccaa gtgggtatgc 4321 gctatgcaat gttaagctag gtcaggtcag accaggtcca aggacagcaa cttgactgta 4381 tgcaaccttt accatctttg cacagaacat acttgtagct agctagttac acttatggac 4441 cgaaaaggca ccccaccatg tctgtccggc tttagagtac ggccgcagac cgctgatttg 4501 ccttgccaag cagtagtcac aatgcatcgc atgagcacac gggcacgggc acgggcacag 4561 gaaccattgg caaaaatacc agatacacta taccgacgta tatcaagccc aagtttaaaa 4621 ttcctaaatt tccgcggcta cttttcaatt ccctatagtg agtcgtatta aattcgtaat 4681 catgtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg 4741 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat 4801 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg 4861 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 4921 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 4981 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 5041 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 5101 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 5161 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 5221 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 5281 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 5341 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 5401 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 5461 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 5521 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 5581 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 5641 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 5701 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 5761 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 5821 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 5881 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 5941 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 6001 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 6061 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 6121 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 6181 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 6241 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 6301 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 6361 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 6421 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 6481 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 6541 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 6601 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 6661 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 6721 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 6781 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc 6841 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac 6901 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt 6961 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc 7021 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc 7081 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact 7141 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg 7201 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc 7261 gaattttaac aaaatattaa cgcttacaat ttccattcgc cattcaggct gcgcaactgt 7321 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt 7381 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg 7441 acggccagtg ccaagctccc gcggggatcg actcataaaa tagtaacctt ctaatgcgta 7501 tctattgact accaaccatt agtgtggttg cagaaggcgg aattctccct tcttcgaatt 7561 cagcttgctt tttcattttt tattttccat ttttcagttt ttgtttgtgt cgaatttagc 7621 cagttgcttc tccaagatga aaaaaacccc tgcgcagttt ctgtgctgca agatcctaat 7681 cgacttttcc accccccaca aaagtaaatg ttcttttgtt acattcgcgt gggtagctag 7741 ctccccgaat cttcaaagga cttagggact gcactacatc agagtgtgtt cacctggttt 7801 gctgcctggt ttgaaagaaa agagcaggga actcgcgggt tcccggcgaa taatcatgcg 7861 atagtccttt ggccttccaa gtcgcatgta gagtagacaa cagacaggga gggcaggaag 7921 gatctttcac tgagatcctg tatcttgttg ggtaagtcgg atgaaagggg aatcgtatga 7981 gattggagag gatgcggaag aggtaacgcc ttttgttaac ttgtttaatt attatggggc 8041 aggcgagagg gggaggaatg tatgtgtgtg aggcgggcga gacggagcca tccaggccag 8101 gtagaaatag agaaagccga atgttagaca atatggcagc gtagtagagt aggtaggtag 8161 gcaagtactg ctagcaaaga ggagaagggt aagctcactc ttcgcattcc acaccgttag 8221 tgtgtcagtt tgaacaaaaa aacaatcatc ataccaattg atggactgtg gactggcttt 8281 tggaacggct tttcggactg cgattattcg tgaggaatca aggtaggaat ttggtcatat 8341 ttacggacaa cagtgggtga ttcccatatg gagtaggaaa acgagatcat ggtatcctca 8401 gatatgttgc ggaattctgt tcaccgcaaa gttcagggtg ctctggtggg tttcggttgg 8461 tctttgcttt gcttctccct tgtcttgcat gttaataata gcctagcctg tgagccgaaa 8521 cttagggtag gcttagtgtt ggaacgtaca tatgtatcac gttgacttgg tttaaccagg 8581 cgacctggta gccagccata cccacacacg ttttttgtat cttcagtata gttgtgaaaa 8641 gtgtagcgga aatttgtggt ccgagcaaca gcgtcttttt ctagtagtgc ggtcggttac 8701 ttggttgaca ttggtatttg gactttgttg ctacaccatt cactacttga agtcgagtgt 8761 gaagggtatg atttctagtg gtgaacacct ttagttacgt aatgttttca ttgctgtttt 8821 acttgagatt tcgattgaga aaaaggtatt taatagctcg aatcaatgtg agaacagaga 8881 gaagatgttc ttccctaact cgaaaggtat atgaggcttg tgtttcttag gagaattatt 8941 attcttttgt tatgttgcgc ttgtagttgg aaaaggtgaa gagacaaaag ctggaattgt 9001 gagcggataa caagctcaac acttgaaatt taggaaagag cagaatttgg caaaaaaaat 9061 aaaaaaaaaa taaacacaca tactcatcga g //