pET-32 EK/LIC

pET32 EK/LIC载体基本信息

pET32 EK/LIC载体质粒图谱和多克隆位点信息

pET32 EK/LIC载体简介

The pET-32 Ek/LIC vector is prepared for rapid, directional cloning of PCR-amplified DNA for high-level expression of polypeptides fused with the 109aa Trx•Tag™ thioredoxin protein. Using specifically designed primers for amplification and the pET-32 Ek/LIC Cloning Kit, inserts can be efficiently cloned without the need for restriction digestion or ligation. Fusion proteins also contain cleavable His•Tag® and S•Tag™ sequences for detection and purification. 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.

pET32 EK/LIC载体序列

ORIGIN
    1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
   61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
  121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
  181 cgacggagct cgaattcgga tccgatatcg ccatggttga ggagaagccc gggctcttgt
  241 cgtcgtcatc ggtacccaga tctgggctgt ccatgtgctg gcgttcgaat ttagcagcag
  301 cggtttcttt cataccagaa ccgcgtggca ccagaccaga agaatgatga tgatgatggt
  361 gcatatggcc agaaccagaa ccggccaggt tagcgtcgag gaactctttc aactgacctt
  421 tagacagtgc acccactttg gttgccgcca cttcaccgtt tttgaacagc agcagagtcg
  481 ggataccacg gatgccatat ttcggcgcag tgccagggtt ttgatcgatg ttcagttttg
  541 caacggtcag tttgccctga tattcgtcag cgatttcatc cagaatcggg gcgatcattt
  601 tgcacggacc gcaccactct gcccagaaat cgacgaggat cgccccgtcc gctttgagta
  661 catccgtgtc aaaactgtcg tcagtcaggt gaataatttt atcgctcata tgtatatctc
  721 cttcttaaag ttaaacaaaa ttatttctag aggggaattg ttatccgctc acaattcccc
  781 tatagtgagt cgtattaatt tcgcgggatc gagatcgatc tcgatcctct acgccggacg
  841 catcgtggcc ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat
  901 caccgatggg gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg
  961 tatggtggca ggccccgtgg ccgggggact gttgggcgcc atctccttgc atgcaccatt
 1021 ccttgcggcg gcggtgctca acggcctcaa cctactactg ggctgcttcc taatgcagga
 1081 gtcgcataag ggagagcgtc gagatcccgg acaccatcga atggcgcaaa acctttcgcg
 1141 gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa
 1201 cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga
 1261 accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc
 1321 tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg
 1381 gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat
 1441 ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg
 1501 aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta
 1561 actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg
 1621 cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag
 1681 acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt
 1741 tagcgggccc attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc
 1801 tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg
 1861 gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg
 1921 ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg
 1981 gtgcggacat ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc
 2041 cgttaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc
 2101 tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga
 2161 aaagaaaaac caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
 2221 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
 2281 attaatgtaa gttagctcac tcattaggca ccgggatctc gaccgatgcc cttgagagcc
 2341 ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg
 2401 actgtcttct ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc
 2461 ggcgaggacc gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga
 2521 atcttgcacg ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag
 2581 aagcaggcca ttatcgccgg catggcggcc ccacgggtgc gcatgatcgt gctcctgtcg
 2641 ttgaggaccc ggctaggctg gcggggttgc cttactggtt agcagaatga atcaccgata
 2701 cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc aacaacatga
 2761 atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc gccctgcacc
 2821 attatgttcc ggatctgcat cgcaggatgc tgctggctac cctgtggaac acctacatct
 2881 gtattaacga agcgctggca ttgaccctga gtgatttttc tctggtcccg ccgcatccat
 2941 accgccagtt gtttaccctc acaacgttcc agtaaccggg catgttcatc atcagtaacc
 3001 cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta cccccatgaa cagaaatccc
 3061 ccttacacgg aggcatcagt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt
 3121 atcagaagcc agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag
 3181 gcagacatct gtgaatcgct tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg
 3241 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
 3301 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
 3361 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta
 3421 actatgcggc atcagagcag attgtactga gagtgcacca tatatgcggt gtgaaatacc
 3481 gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga
 3541 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
 3601 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
 3661 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
 3721 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
 3781 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
 3841 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
 3901 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
 3961 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
 4021 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
 4081 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
 4141 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
 4201 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
 4261 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
 4321 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
 4381 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
 4441 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
 4501 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
 4561 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
 4621 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
 4681 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
 4741 agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc
 4801 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
 4861 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
 4921 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
 4981 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
 5041 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
 5101 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
 5161 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
 5221 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
 5281 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
 5341 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
 5401 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg aaattgtaaa
 5461 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
 5521 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
 5581 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
 5641 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
 5701 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
 5761 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
 5821 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
 5881 gcttaatgcg ccgctacagg gcgcgtccca ttcgcca
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