表达载体pHAT2图谱及序列信息

表达载体pHAT2图谱及序列信息,该载体含有N-His标签

EMBL Protein Expression and Purification Facility

pHAT2 - His-tagged expression vector

Features

T7 promoter

Lac operator

T7 gene 10 ribosome binding site

His-tag

Polylinker

SP6 promoter

T7 transcription terminator

LacI gene

β-Lactamase gene

Origin of replicationposition1-1721-4250-8092-109111-179184-202281-322732-1813 (reversed)2583-34333507-4203

Reference:

Peränen J., Rikkonen M.,Hyvönen M. & Kääriäinen L. (1996) T7 vectors with modified T7lac promoter forexpression of proteins in Escherichia coli. Anal. Biochem. 236

, 371-373.

表达载体pHAT2图谱及序列信息,该载体含有N-His标签

pHAT2

T7 promoter --> Lac operator XbaI

ATTAATACGACTCACTATAGGGAATTGTGAGCGGATAACAATTCCTCTAGAAATAATTTTTAATTATGCTGAGTGATATCCCTTAACACTCGCCTATTGTTAAGGAGATCTTTATTAAAA rbs His-tag NcoIATGCAAATTGAAATTCTTCCTCTATATAGGTACTCAGTAGTGGTAGTGGTAGTGAGGTACCTA METSerHisHisHsiHisHisHisSerMetAsp EcoRI SalI XhoI BamHI NotI NsiI HindIIIATCGAATTCGTCGACCTCGAGGGATCCGGGCCCTCTAGATGCGGCCGCATGCATAAGCTTTAGCTTAAGCAGCTGGAGCTCCCTAGGCCCGGGAGATCTACGCCGGCGTACGTATTCGAAIleGluPheValAspLeuGluGlySerGlyProSerArgCysGlyArgMetHisLysLeu <-- SP6 promoter

GAGTATTCTATAGTGTCACCTAAATCCCAGCTTGATCCGGCTGCTAACAAAGCCCGAAAGCTCATAAGATATCACAGTGGATTTAGGGTCGAACTAGGCCGACGATTGTTTCGGGCTTTCGluTyrSerIleValSerProLysSerGlnLeuAspProAlaAlaAsnLysAlaArgLys111 NcoI 173 HindIII 1277 BstEII 2014 Eco56I122 EcoRI 264 EspI 1445 BclI 2016 NaeI128 SalI 264 Bpu1102I 1459 MluI 2196 NdeI134 XhoI 341 AccIII 1884 PflMI 2451 AatII136 SciI 341 BspMII 1894 Asp718I 2770 XmnI140 BamHI 650 BspMI 1894 Acc65I 2889 ScaI160 NotI 661 PstI 1898 KpnI 3304 Eco31I160 EagI 821 NarI 1902 Ecl136II 3370 Eam1105I160 Eco52I 822 EheI 1902 EcoICRI 3849 AlwNI160 XmaIII 824 BbeI 1904 SacI 4375 SapI171 NsiI 1048 BssHII 1904 SstIAflII AgeI AscI AsuII AvrII BalI

BglII BsmI Bsp1407I Bsu36I ClaI Csp45I

CspI CvnI DraIII Eco47III Eco72I EcoNI

FseI HpaI I-PpoI MfeI Mlu113I MscI

MstII NheI NruI PacI PinAI PmaCI

PmeI RleAI SacII SauI SfiI SgrAI

SmaI SnaBI SpeI SplI SpoI SrfISstII StuI SunI SwaI Tth111I XmaI

表达载体pHAT2图谱及序列信息,该载体含有N-His标签

LOCUS pHAT2 4443 bp DNA circular 29-NOV-2006SOURCE ORGANISM COMMENT This file is created by Vector NTI /COMMENT ORIGDB|GenBankCOMMENT VNTDATE|428249734|COMMENT VNTDBDATE|428368939|COMMENT LSOWNER|COMMENT VNTNAME|pHAT2|COMMENT VNTAUTHORNAME|Hüseyin Besir|COMMENT VNTOAUTHORNAME|UNKNOWN|COMMENT VNTREPLTYPE|PlasmidCOMMENT VNTEXTCHREPL|BacteriaFEATURES Location/Qualifiers promoter 1..19 /gene="T7 prom" /vntifkey="30" /label=T7\prom /note="Promoter" terminator 219..347 /gene="T7 term" /vntifkey="43" /label=T7\term /note="Terminator" CDS complement(732..1814) /sequence="ORF_3 rf(5)" /vntifkey="4" /label=lacI promoter 2513..2541 /gene="amp prom" /vntifkey="30" /label=amp\prom /note="Promoter" CDS 2583..3443 /sequence="ORF_2 rf(3)" /vntifkey="4" /label=ampR /note="ampR" rep_origin 3598..4217 /gene="pBR322 origin" /vntifkey="33" /label=pBR322\origin /note="Rep_Origin" misc_binding 92..109 /vntifkey="20" /label=His-tagBASE COUNT 1097 a 1139 c 1144 g 1063 t ORIGIN 1 taatacgact cactataggg aattgtgagc ggataacaat tcctctagaa ataattttgt 61 ttaactttaa gaaggagata tatccatgag tcatcaccat caccatcact ccatggatat 121 cgaattcgtc gacctcgagg gatccgggcc ctctagatgc ggccgcatgc ataagcttga 181 gtattctata gtgtcaccta aatcccagct tgatccggct gctaacaaag cccgaaagga 241 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa 301 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggataac ctggcgtaat 361 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg

表达载体pHAT2图谱及序列信息,该载体含有N-His标签

421 aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa tatttgttaa atcagctcat 481 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga 541 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca 601 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacagctt gcatgcctgc 661 aggtcggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa 721 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat 781 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt ggtttttctt 841 ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg agagagttgc 901 agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat ggtggttgac 961 ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga gatatccgca 1021 ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg 1081 gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt ttgttgaaaa 1141 ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg attgcgagtg 1201 agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa tgggcccgct 1261 aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag tcgcgtaccg 1321 tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc aagaaataac 1381 gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc cagcggatag 1441 ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc tttacaggct 1501 tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg atcggcgcga 1561 gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg 1621 ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg aatgtaattc 1681 agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg gctggcctgg 1741 ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac atcgtataac 1801 gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta tcatgccata 1861 ccgcgaaagg ttttgcacca ttcgatggtg tcgggtaccg agctcgaatt tacgtgaacc 1921 atcacccaaa tcaagttttt tgcggtcgag gtgccgtaaa gctctaaatc ggaaccctaa 1981 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg 2041 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt 2101 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcgcctg atgcggtatt 2161 ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct 2221 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct 2281 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct 2341 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga 2401 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca 2461 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata 2521 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga 2581 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc 2641 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg 2701 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc 2761 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtcat acactattat 2821 cccgtattga cgccgggcaa gagcaactcg gtcgccgggc gcggtattct cagaatgact 2881 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat 2941 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga 3001 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc 3061 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga 3121 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag 3181 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc 3241 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt 3301 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct 3361 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg 3421 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg 3481 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca 3541 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga 3601 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa 3661 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga 3721 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt 3781 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt

表达载体pHAT2图谱及序列信息,该载体含有N-His标签

3841 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat 3901 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct 3961 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca 4021 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag 4081 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc 4141 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga 4201 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca 4261 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag 4321 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg 4381 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct 4441 gat //

Full length: 4443 bp

Composition: 1097 A; 1139 C; 1144 G; 1063 T; 0 OTHER

Percentage: 25% A; 26% C; 26% G; 24% T; 0%OTHER

Molecular Weight (kDa): ssDNA: 1372.06 dsDNA: 2739.2

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