pET-25b (Cat. No. 69753-3)载体带有pelB信号肽序列,能够将目的蛋白定位在细菌周质腔,还带有C端HSV和His标签。pET25b载体的单一的多克隆位点见上面的环状质粒图谱。注意:载体序列是以pBR322质粒的编码规矩进行编码的,所以T7蛋白表达区在质粒图谱上面是反向的。T7 RNA聚合酶启动的克隆和表达区域在质粒图谱中也被标注了出来。质粒的F1复制子是被定向的,所以在T7噬菌体聚合酶的作用下,包含有蛋白编码序列的病毒 粒子能够产生,并启动蛋白表达,同时蛋白表达将被T7终止子序列(Cat. No. 69337-3)的作用下终止蛋白翻译。
LOCUS pET-25b(+) 5547 bp DNA circular SYNDEFINITION pET-25b(+)ACCESSION KEYWORDS SOURCE ORGANISM other sequences; artificial sequences; vectors.COMMENT This file is created by Vector NTI http://www.biofeng.com/COMMENT ORIGDB|GenBankCOMMENT VNTAUTHORNAME|biofeng.com|COMMENT VNTOAUTHORAD1|Novagen Inc.|FEATURES Location/Qualifiers source 1..5547 /organism="pET-25b(+)" /mol_type="other DNA" terminator complement(1..129) /label="T7_terminator" misc_feature 69..87 /label="T7_Terminal_primer" misc_feature complement(352..368) /label="T7_transl_en_RBS" misc_feature complement(386..413) /label="lacO" promoter complement(413..431) /label="T7_promoter" gene 463..726 /label="tet (300 - 563)" /gene="tet (300 - 563)" misc_feature complement(515..534) /label="pBRrevBam_primer" CDS 568..1068 /label="ORF frame 1" misc_feature 809..1900 /label="lacI" CDS 941..1900 /label="ORF frame 2" gene 1959..2234 /label="tet (576 - 851)" /gene="tet (576 - 851)" misc_feature 2709..2900 /label="ROP" misc_feature complement(2894..2916) /label="pGEX_3_primer" rep_origin complement(3315..3934) /label="pBR322_origin" gene complement(4089..4949) /label="Ampicillin" /gene="Ampicillin" CDS complement(4089..4949) /label="ORF frame 2" promoter complement(4991..5019) /label="AmpR_promoter"ORIGIN 1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa 61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt 121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gacatcctcg gggtcttccg 181 gggcgagttc tggctggcta gcccgtttga tctcgagtgc ggccgcaagc ttgtcgacgg 241 agctcgaatt cggatccgaa ttaattccga tatccatggc catcgccggc tgggcagcga 301 ggagcagcag accagcagca gcggtcggca gcaggtattt catatgtata tctccttctt 361 aaagttaaac aaaattattt ctagagggga attgttatcc gctcacaatt cccctatagt 421 gagtcgtatt aatttcgcgg gatcgagatc tcgatcctct acgccggacg catcgtggcc 481 ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat caccgatggg 541 gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg tatggtggca 601 ggccccgtgg ccgggggact gttgggcgcc atctccttgc atgcaccatt ccttgcggcg 661 gcggtgctca acggcctcaa cctactactg ggctgcttcc taatgcagga gtcgcataag 721 ggagagcgtc gagatcccgg acaccatcga atggcgcaaa acctttcgcg gtatggcatg 781 atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga 841 tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag 901 ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat 961 tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac 1021 ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga 1081 tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa 1141 agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct 1201 ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct 1261 tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg 1321 actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc 1381 attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa 1441 tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca 1501 aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca 1561 gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat 1621 ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac 1681 catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc 1741 tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac 1801 caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca 1861 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtaa 1921 gttagctcac tcattaggca ccgggatctc gaccgatgcc cttgagagcc ttcaacccag 1981 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct 2041 ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc ggcgaggacc 2101 gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg 2161 ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag aagcaggcca 2221 ttatcgccgg catggcggcc ccacgggtgc gcatgatcgt gctcctgtcg ttgaggaccc 2281 ggctaggctg gcggggttgc cttactggtt agcagaatga atcaccgata cgcgagcgaa 2341 cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc aacaacatga atggtcttcg 2401 gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc gccctgcacc attatgttcc 2461 ggatctgcat cgcaggatgc tgctggctac cctgtggaac acctacatct gtattaacga 2521 agcgctggca ttgaccctga gtgatttttc tctggtcccg ccgcatccat accgccagtt 2581 gtttaccctc acaacgttcc agtaaccggg catgttcatc atcagtaacc cgtatcgtga 2641 gcatcctctc tcgtttcatc ggtatcatta cccccatgaa cagaaatccc ccttacacgg 2701 aggcatcagt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt atcagaagcc 2761 agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag gcagacatct 2821 gtgaatcgct tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg cgtttcggtg 2881 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag 2941 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg 3001 gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc 3061 atcagagcag attgtactga gagtgcacca tatatgcggt gtgaaatacc gcacagatgc 3121 gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc 3181 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc 3241 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg 3301 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 3361 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 3421 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 3481 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 3541 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 3601 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 3661 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 3721 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt 3781 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 3841 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 3901 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 3961 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 4021 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 4081 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 4141 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 4201 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 4261 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 4321 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 4381 ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg 4441 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 4501 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 4561 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 4621 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 4681 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta 4741 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 4801 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 4861 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 4921 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 4981 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 5041 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg aaattgtaaa cgttaatatt 5101 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa 5161 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca 5221 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc 5281 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg 5341 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg 5401 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg 5461 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg 5521 ccgctacagg gcgcgtccca ttcgcca