| 出品公司: | NEB |
|---|---|
| 载体名称: | pKLAC1 |
| 质粒类型: | 乳酸酵母表达系统 |
| 高拷贝/低拷贝: | -- |
| 克隆方法: | 多克隆位点，限制性内切酶 |
| 启动子: | LAC4 |
| 载体大小: | 9091dp |
| 5' 测序引物及序列: | -- |
| 3' 测序引物及序列: | -- |
| 载体标签: | -- |
| 载体抗性: | 氨苄青霉素 |
| 筛选标记: | -- |
| 克隆菌株: | -- |
| 宿主细胞（系）: | -- |
| 备注: | -- |
| 产品目录号: | -- |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
pKLAC1是一种既能在大肠杆菌中复制又能稳定整合到酵母Kluyveromyces lactis（1）基因组中的表达载体。它是为利用乳双歧杆菌蛋白表达试剂盒（NEB#E1000）在乳双歧杆菌中高水平表达重组蛋白而设计的。
载体序列
LOCUS pKLAC1 9091 bp ds-DNA circular SYN 29-NOV-2013
DEFINITION Integrative expression vector for the yeast Kluyveromyces lactis.
For an alternative MCS, use pKLAC2.
ACCESSION AY968582
VERSION .
KEYWORDS pKLAC1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9091)
AUTHORS Colussi PA, Taron CH.
TITLE Kluyveromyces lactis LAC4 promoter variants that lack function in
bacteria but retain full function in K. lactis.
JOURNAL Appl. Environ. Microbiol. 2005;71:7092-8.
PUBMED 16269745
REFERENCE 2 (bases 1 to 9091)
AUTHORS New England Biolabs
TITLE Direct Submission
JOURNAL Exported Saturday, November 9, 2019 from SnapGene Viewer 3.1.4
http://www.snapgene.com
COMMENT Linearize with SacII or BstXI to integrate at the LAC4 locus.
FEATURES Location/Qualifiers
source 1..9091
/organism="synthetic DNA construct"
/lab_host="Kluyveromyces lactis"
/mol_type="other DNA"
CDS 14..280
/codon_start=1
/product="N-terminal secretion signal from Kluyveromyces
lactis alpha-factor"
/note="alpha-factor secretion signal"
/note="Cleavage by the Kex2 protease occurs after the
dibasic KR sequence. The EA dipeptides are then removed by
dipeptidyl aminopeptidase A."
/translation="MKFSTILAASTALISVVMAAPVSTETDIDDLPISVPEEALIGFID
LTGDEVSLLPVNNGTHTGILFLNTTIAEAAFADKDDLEKREAEA"
misc_feature 257..343
/note="MCS"
/note="multiple cloning site"
terminator 355..973
/note="LAC4 terminator"
/note="transcription terminator for Kluyveromyces lactis
LAC4"
promoter 974..1676
/gene="S. cerevisiae ADH1"
/note="ADH1 promoter"
/note="promoter for alcohol dehydrogenase 1"
CDS 1697..3343
/codon_start=1
/product="acetamidase from Aspergillus nidulans"
/note="amdS"
/note="confers ability to process acetamide for selection
in fungal cells"
/translation="MPQSWEELAADKRARLAKTIPDEWKVQTLPAEDSVIDFPKKSGIL
SEAELKITEASAADLVSKLAAGELTSVEVTLAFCKRAAIAQQLTNCAHEFFPDAALAQA
RELDEYYAKHKRPVGPLHGLPISLKDQLRVKGYETSMGYISWLNKYDEGDSVLTTMLRK
AGAVFYVKTSVPQTLMVCETVNNIIGRTVNPRNKNWSCGGSSGGEGAIVGIRGGVIGVG
TDIGGSIRVPAAFNFLYGLRPSHGRLPYAKMANSMEGQETVHSVVGPITHSVEDLRLFT
KSVLGQEPWKYDSKVIPMPWRQSESDIIASKIKNGGLNIGYYNFDGNVLPHPPILRGVE
TTVAALAKAGHTVTPWTPYKHDFGHDLISHIYAADGSADVMRDISASGEPAIPNIKDLL
NPNIKAVNMNELWDTHLQKWNYQMEYLEKWREAEEKAGKELDAIIAPITPTAAVRHDQF
RYYGYASVINLLDFTSVVVPVTFADKNIDKKNESFKAVSELDALVQEEYDPEAYHGAPV
AVQVIGRRLSEERTLAIAEEVGKLLGNVVTP"
promoter 3979..4635
/note="LAC4 promoter (5')"
/note="5' portion of a modified Kluyveromyces lactis LAC4
promoter"
rep_origin complement(5086..5674)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5845..6705)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(6706..6810)
/gene="bla"
/note="AmpR promoter"
rep_origin 6837..7292
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 7463..8991
/note="LAC4 promoter (3')"
/note="3' portion of a modified Kluyveromyces lactis LAC4
promoter"
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
//
