pET-41 EK/LIC

pET-41 EK/LIC

pET41 EK/LIC载体基本信息

出品公司:	Ybscience
别名:	pET41 EK/LIC, pET 41 EK/LIC
质粒类型:	大肠杆菌蛋白表达
表达水平:	高
克隆方法:	多克隆位点，限制性内切酶
载体大小:	5947 bp
5' 测序引物:	T7或pGEX-5‘
5' 测序引物序列:	T7: 5'-TAATACGACTCACTATAGGG-3'; pGEX-5': 5'-GGGCTGGCAAGCCACGTTTGGTG-3'
载体标签:	N-GST, N-His, N-EK, C-His
载体抗性:	Kanamycin
备注:	For directional cloning of PCR-amplified DNA; ligation independent cloning; enterokinase cleavage site.
稳定性:	瞬时表达 Transient
组成型:	组成型 Constitutive
病毒/非病毒:	非病毒

 

pET41 EK/LIC载体质粒图谱和多克隆位点信息

pET41 EK/LIC载体简介

The pET-41 Ek/LIC vector is prepared for rapid, directional cloning of PCR-amplified DNA for high-level expression of polypeptides fused with N-terminal GST•Tag™, His•Tag® and S•Tag™ sequences. Using specifically designed primers for amplification and the pET-41 Ek/LIC Vector Kit (Cat. No. 71071-3), inserts can be efficiently cloned without the need for restriction digestion or ligation. Unique sites are shown on the circle map. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the circle map. The cloning/expression region of the coding strand transcribed by T7 RNA polymerase is shown below. The f1 origin is oriented so that infection with helper phage will produce virions containing single stranded DNA that corresponds to the coding strand. Therefore, single stranded sequencing should be performed using the T7 terminator primer (Cat. No. 69337-3). Vector encoded sequence can be completely removed when cloning into the Ek/LIC site (as shown below left) by cleaving the fusion protein with enterokinase.

pET41 EK/LIC载体序列

LOCUS       pET-41 Ek/LIC    5947 bp     DNA  Circular    SYN
DEFINITION  pET-41 Ek/LIC
ACCESSION   
KEYWORDS    
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
COMMENT     This file is created by Vector NTI
            http://www.biofeng.com/
COMMENT     ORIGDB|GenBank
COMMENT     VNTAUTHORNAME|biofeng.com|
COMMENT     VNTOAUTHORAD1|Novagen Inc.|
FEATURES             Location/Qualifiers
     source          1..5947
                     /organism="pET-41 Ek/LIC"
                     /mol_type="other DNA"
     terminator      complement(1..120)
                     /label="T7_terminator"
     misc_feature    69..87
                     /label="T7_Terminal_primer"
     CDS             complement(147..1109)
                     /label="ORF frame 3"
     misc_feature    complement(279..293)
                     /label="EK"
     misc_feature    complement(324..368)
                     /label="S15"
     misc_feature    complement(411..428)
                     /label="6xHIS"
     gene            complement(459..1109)
                     /label="GST (variant)"
                     /gene="GST (variant)"
     CDS             1110..1838
                     /label="ORF frame 3"
     misc_feature    complement(1118..1134)
                     /label="T7_transl_en_RBS"
     misc_feature    complement(1152..1179)
                     /label="lacO"
     promoter        complement(1179..1197)
                     /label="T7_promoter"
     gene            1233..1496
                     /label="tet (300 - 563)"
                     /gene="tet (300 - 563)"
     misc_feature    complement(1285..1304)
                     /label="pBRrevBam_primer"
     misc_feature    1579..2670
                     /label="lacI"
     CDS             1711..2670
                     /label="ORF frame 1"
     misc_feature    3242..3433
                     /label="ROP"
     misc_feature    complement(3427..3449)
                     /label="pGEX_3_primer"
     rep_origin      complement(3848..4467)
                     /label="pBR322_origin"
     gene            4573..5388
                     /label="KanR2"
                     /gene="KanR2"
     CDS             4573..5388
                     /label="ORF frame 1"
ORIGIN
    1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
   61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
  121 tgtttagcag cctaggtatt aatcaattag tggtggtggt ggtggtggtg gtgctcgagt
  181 gcggccgcaa gcttgtcgac ggagctcgcc tgcaggcgcg ccaaggcctg tacagaattc
  241 ggatccccga tatccatggt tgaggagaag cccgggctct tgtcgtcgtc atcaccagaa
  301 ccaccaccgg tacccagatc tgggctgtcc atgtgctggc gttcgaattt agcagcagcg
  361 gtttctttca taccaattgc agtactaccg cgtggcacca gacccgcgga gtgatggtga
  421 tggtgatgac cagaaccact agttgaacca tccgattttg gaggatggtc gccaccacca
  481 aacgtggctt gccagccctg caaaggccat gctatatact tgctggattt caagtactta
  541 tcaatttgtg ggatagcttc aatacgtttt ttaaaacaaa ctaattttgg gaacgcatcc
  601 aggcacattg ggtccatgta taaaacaaca tcaagagcgt catacaacat gaagtcagga
  661 tgggttacat gatcaccatt taaatatgtt ttatgacata aacgatcttc gaacattttc
  721 agcatttcag gtagcttgct aagaaaatca actttgagag tttcaaagtc tttactatat
  781 gcaattctcg aaacaccgta tctaatatcc aaaaccgctc cttcaagcat tgaaatctct
  841 gcacgctctt ttggacaacc acccaacatg ttgtgcttgt cagctatata acgtatgatg
  901 gccatagact gtgttaattt aacatcacca tcaatataat aaggaagatt gggaaactcc
  961 aaacccaatt caaacttttt gtttcgccat ttatcacctt catcgcgctc atacaaatgc
 1021 tcttcatatt tttcttcaag atattccaaa agaagtcgag tgggttgcac aaggccctta
 1081 attttccaat aacctagtat aggggacata tgtatatctc cttcttaaag ttaaacaaaa
 1141 ttatttctag aggggaattg ttatccgctc acaattcccc tatagtgagt cgtattaatt
 1201 tcgcgggatc gagatcgatc tcgatcctct acgccggacg catcgtggcc ggcatcaccg
 1261 gcgccacagg tgcggttgct ggcgcctata tcgccgacat caccgatggg gaagatcggg
 1321 ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg
 1381 ccgggggact gttgggcgcc atctccttgc atgcaccatt ccttgcggcg gcggtgctca
 1441 acggcctcaa cctactactg ggctgcttcc taatgcagga gtcgcataag ggagagcgtc
 1501 gagatcccgg acaccatcga atggcgcaaa acctttcgcg gtatggcatg atagcgcccg
 1561 gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag
 1621 tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct
 1681 gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc
 1741 gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg
 1801 gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt
 1861 gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg
 1921 cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag
 1981 gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct
 2041 gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg
 2101 gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct
 2161 gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag
 2221 ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa
 2281 atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg
 2341 ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggacat ctcggtagtg
 2401 ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag
 2461 gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag
 2521 gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg
 2581 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
 2641 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtaa gttagctcac
 2701 tcattaggca ccgggatctc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt
 2761 ccggtgggcg cggggcatga ctagcatgat cgtgctcctg tcgttgagga cccggctagg
 2821 ctggcggggt tgccttactg gttagcagaa tgaatcaccg atacgcgagc gaacgtgaag
 2881 cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca tgaatggtct tcggtttccg
 2941 tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc accattatgt tccggatctg
 3001 catcgcagga tgctgctggc taccctgtgg aacacctaca tctgtattaa cgaagcgctg
 3061 gcattgaccc tgagtgattt ttctctggtc ccgccgcatc cataccgcca gttgtttacc
 3121 ctcacaacgt tccagtaacc gggcatgttc atcatcagta acccgtatcg tgagcatcct
 3181 ctctcgtttc atcggtatca ttacccccat gaacagaaat cccccttaca cggaggcatc
 3241 agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc tttatcagaa gccagacatt
 3301 aacgcttctg gagaaactca acgagctgga cgcggatgaa caggcagaca tctgtgaatc
 3361 gcttcacgac cacgctgatg agctttaccg cagctgcctc gcgcgtttcg gtgatgacgg
 3421 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
 3481 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc
 3541 catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag
 3601 cagattgtac tgagagtgca ccatatatgc ggtgtgaaat accgcacaga tgcgtaagga
 3661 gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
 3721 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
 3781 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
 3841 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
 3901 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
 3961 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
 4021 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
 4081 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
 4141 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
 4201 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
 4261 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
 4321 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
 4381 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
 4441 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
 4501 actcacgtta agggattttg gtcatgaaca ataaaactgt ctgcttacat aaacagtaat
 4561 acaaggggtg ttatgagcca tattcaacgg gaaacgtctt gctctaggcc gcgattaaat
 4621 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
 4681 ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
 4741 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg
 4801 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
 4861 ctcaccactg cgatccccgg gaaaacagca ttccaggtat tagaagaata tcctgattca
 4921 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt
 4981 tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg
 5041 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
 5101 caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat
 5161 ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat
 5221 gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc
 5281 ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct
 5341 gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaaga attaattcat
 5401 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
 5461 tccccgaaaa gtgccacctg aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa
 5521 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
 5581 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
 5641 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
 5701 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
 5761 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
 5821 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
 5881 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca
 5941 ttcgcca