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JOURNAL Unpublished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
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