| 质粒名称: | pAKTaq |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 25712 |
| 存储实验室: | David Engelke |
|
载体骨架基本信息 |
|
| 载体名称: | pTTQ18 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4563 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Taq polymerase |
| 物种来源: | Thermus aquaticus |
| 插入大小: | 2501 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | EcoRI（未销毁） |
| 3' 克隆位点: | BglII（销毁后未知） |
| 5' 测序引物 | Tac promoter |
| 3' 测序引物 | M13_puc_F |
|
文献引用方法 |
|
| 材料和方法部分: | pAKTaq was a gift from David Engelke (Addgene plasmid # 25712 ; http://n2t.net/addgene:25712 ; RRID:Addgene_25712) |
| 参考文献部分: | Purification of Thermus aquaticus DNA polymerase expressed in Escherichia coli. Engelke DR, Krikos A, Bruck ME, Ginsburg D. Anal Biochem. 1990 Dec . 191(2):396-400. 10.1016/0003-2697(90)90238-5 PubMed 2085185 |
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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
载体序列
LOCUS pAKTaq 7043 bp ds-DNA circular SYN 02-NOV-2020
DEFINITION Bacterial expression of Taq polymerase.
ACCESSION .
VERSION .
KEYWORDS pAKTaq
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7043)
AUTHORS Engelke DR, Krikos A, Bruck ME, Ginsburg D
TITLE Purification of Thermus aquaticus DNA polymerase expressed in
Escherichia coli.
JOURNAL Anal Biochem. 1990 Dec . 191(2):396-400.
PUBMED 2085185
REFERENCE 2 (bases 1 to 7043)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Nov 2, 2020 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7043
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 131..159
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 167..183
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 172..194
/label=M13/pUC Reverse
/note="In lacZ gene"
CDS 212..2704
/codon_start=1
/product="thermostable DNA polymerase from Thermus
aquaticus"
/label=Taq polymerase
/translation="GMLPLFEPKGRVLLVDGHHLAYRTFHALKGLTTSRGEPVQAVYGF
AKSLLKALKEDGDAVIVVFDAKAPSFRHEAYGGYKAGRAPTPEDFPRQLALIKELVDLL
GLARLEVPGYEADDVLASLAKKAEKEGYEVRILTADKDLYQLLSDRIHVLHPEGYLITP
AWLWEKYGLRPDQWADYRALTGDESDNLPGVKGIGEKTARKLLEEWGSLEALLKNLDRL
KPAIREKILAHMDDLKLSWDLAKVRTDLPLEVDFAKRREPDRERLRAFLERLEFGSLLH
EFGLLESPKALEEAPWPPPEGAFVGFVLSRKEPMWADLLALAAARGGRVHRAPEPYKAL
RDLKEARGLLAKDLSVLALREGLGLPPGDDPMLLAYLLDPSNTTPEGVARRYGGEWTEE
AGERAALSERLFANLWGRLEGEERLLWLYREVERPLSAVLAHMEATGVRLDVAYLRALS
LEVAEEIARLEAEVFRLAGHPFNLNSRDQLERVLFDELGLPAIGKTEKTGKRSTSAAVL
EALREAHPIVEKILQYRELTKLKSTYIDPLPDLIHPRTGRLHTRFNQTATATGRLSSSD
PNLQNIPVRTPLGQRIRRAFIAEEGWLLVALDYSQIELRVLAHLSGDENLIRVFQEGRD
IHTETASWMFGVPREAVDPLMRRAAKTINFGVLYGMSAHRLSQELAIPYEEAQAFIERY
FQSFPKVRAWIEKTLEEGRRRGYVETLFGRRRYVPDLEARVKSVREAAERMAFNMPVQG
TAADLMKLAMVKLFPRLEEMGARMLLQVHDELVLEAPKERAEAVARLAKEVMEGVYPLA
VPLEVEVGIGEDWLSAKE"
primer_bind complement(2745..2762)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(2745..2761)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2754..2776)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind complement(2992..3009)
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
terminator 3162..3248
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 3340..3367
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
primer_bind complement(3482..3501)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 3601..3623
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(3661..3679)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 3735..3812
/gene="lacI (mutant)"
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
CDS 3813..4895
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=lacI
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind complement(3832..3851)
/label=LacI-R
/note="LacI, reverse primer"
promoter 5008..5112
/gene="bla"
/label=AmpR promoter
CDS 5113..5973
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(5331..5350)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6144..6732
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6633..6652
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6886..6903
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 aagctgtggt atggctgtgc aggtcgtaaa tcactgcata attcgtgtcg ctcaaggcgc
61 actcccgttc tggataatgt tttttgcgcc gacatcataa cggttctggc aaatattctg
121 aaatgagctg ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa
181 caatttcaca caggaaacag cgatgaattc ggggatgctg cccctctttg agcccaaggg
241 ccgggtcctc ctggtggacg gccaccacct ggcctaccgc accttccacg ccctgaaggg
301 cctcaccacc agccgggggg agccggtgca ggcggtctac ggcttcgcca agagcctcct
361 caaggccctc aaggaggacg gggacgcggt gatcgtggtc tttgacgcca aggccccctc
421 cttccgccac gaggcctacg gggggtacaa ggcgggccgg gcccccacgc cggaggactt
481 tccccggcaa ctcgccctca tcaaggagct ggtggacctc ctggggctgg cgcgcctcga
541 ggtcccgggc tacgaggcgg acgacgtcct ggccagcctg gccaagaagg cggaaaagga
601 gggctacgag gtccgcatcc tcaccgccga caaagacctt taccagctcc tttccgaccg
661 catccacgtc ctccaccccg aggggtacct catcaccccg gcctggcttt gggaaaagta
721 cggcctgagg cccgaccagt gggccgacta ccgggccctg accggggacg agtccgacaa
781 ccttcccggg gtcaagggca tcggggagaa gacggcgagg aagcttctgg aggagtgggg
841 gagcctggaa gccctcctca agaacctgga ccggctgaag cccgccatcc gggagaagat
901 cctggcccac atggacgatc tgaagctctc ctgggacctg gccaaggtgc gcaccgacct
961 gcccctggag gtggacttcg ccaaaaggcg ggagcccgac cgggagaggc ttagggcctt
1021 tctggagagg cttgagtttg gcagcctcct ccacgagttc ggccttctgg aaagccccaa
1081 ggccctggag gaggccccct ggcccccgcc ggaaggggcc ttcgtgggct ttgtgctttc
1141 ccgcaaggag cccatgtggg ccgatcttct ggccctggcc gccgccaggg ggggccgggt
1201 ccaccgggcc cccgagcctt ataaagccct cagggacctg aaggaggcgc gggggcttct
1261 cgccaaagac ctgagcgttc tggccctgag ggaaggcctt ggcctcccgc ccggcgacga
1321 ccccatgctc ctcgcctacc tcctggaccc ttccaacacc acccccgagg gggtggcccg
1381 gcgctacggc ggggagtgga cggaggaggc gggggagcgg gccgcccttt ccgagaggct
1441 cttcgccaac ctgtggggga ggcttgaggg ggaggagagg ctcctttggc tttaccggga
1501 ggtggagagg cccctttccg ctgtcctggc ccacatggag gccacggggg tgcgcctgga
1561 cgtggcctat ctcagggcct tgtccctgga ggtggccgag gagatcgccc gcctcgaggc
1621 cgaggtcttc cgcctggccg gccacccctt caacctcaac tcccgggacc agctggaaag
1681 ggtcctcttt gacgagctag ggcttcccgc catcggcaag acggagaaga ccggcaagcg
1741 ctccaccagc gccgccgtcc tggaggccct ccgcgaggcc caccccatcg tggagaagat
1801 cctgcagtac cgggagctca ccaagctgaa gagcacctac attgacccct tgccggacct
1861 catccacccc aggacgggcc gcctccacac ccgcttcaac cagacggcca cggccacggg
1921 caggctaagt agctccgatc ccaacctcca gaacatcccc gtccgcaccc cgcttgggca
1981 gaggatccgc cgggccttca tcgccgagga ggggtggcta ttggtggccc tggactatag
2041 ccagatagag ctcagggtgc tggcccacct ctccggcgac gagaacctga tccgggtctt
2101 ccaggagggg cgggacatcc acacggagac cgccagctgg atgttcggcg tcccccggga
2161 ggccgtggac cccctgatgc gccgggcggc caagaccatc aacttcgggg tcctctacgg
2221 catgtcggcc caccgcctct cccaggagct agccatccct tacgaggagg cccaggcctt
2281 cattgagcgc tactttcaga gcttccccaa ggtgcgggcc tggattgaga agaccctgga
2341 ggagggcagg aggcgggggt acgtggagac cctcttcggc cgccgccgct acgtgccaga
2401 cctagaggcc cgggtgaaga gcgtgcggga ggcggccgag cgcatggcct tcaacatgcc
2461 cgtccagggc accgccgccg acctcatgaa gctggctatg gtgaagctct tccccaggct
2521 ggaggaaatg ggggccagga tgctccttca ggtccacgac gagctggtcc tcgaggcccc
2581 aaaagagagg gcggaggccg tggcccggct ggccaaggag gtcatggagg gggtgtatcc
2641 cctggccgtg cccctggagg tggaggtggg gataggggag gactggctct ccgccaagga
2701 gtgatagatc ctctagagtc gacctgcagg catgcaagct tggcactggc cgtcgtttta
2761 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
2821 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
2881 cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
2941 atttcacacc gcataaattc cctgttttgg cggatgagag aagattttca gcctgataca
3001 gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc
3061 ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag
3121 tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc
3181 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta
3241 ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg
3301 caggacgccc gccataaact gccaggcatc aaattaagca gaaggccatc ctgacggatg
3361 gcctttttgc gtttctacaa actcttcctg tcgtcatatc tacaagccat ccccccacag
3421 atacggtaaa ctagcctcgt ttttgcatca ggaaagcagg gaatttatgg tgcactctca
3481 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
3541 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
3601 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
3661 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
3721 gaggttctgt acccgacacc atcgaatggt gcaaaacctt tcgcggtatg gcatgatagc
3781 gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg
3841 cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg
3901 tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca
3961 accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca
4021 gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac
4081 tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg
4141 cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg
4201 accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg
4261 tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg
4321 gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa
4381 gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa
4441 ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca
4501 tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg
4561 cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg
4621 tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca
4681 aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg
4741 gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc
4801 tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
4861 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtaagttag
4921 ctcactcatt aggcacccca ggctttacac tttatgcttc cgacctgcaa gaacctcacg
4981 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
5041 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
5101 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
5161 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
5221 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
5281 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
5341 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
5401 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
5461 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
5521 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
5581 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
5641 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
5701 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
5761 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
5821 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
5881 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
5941 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
6001 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
6061 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
6121 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
6181 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
6241 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
6301 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
6361 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
6421 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
6481 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
6541 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
6601 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
6661 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
6721 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
6781 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
6841 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
6901 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
6961 taatgcagaa ttaattctca tgtttgacag cttatcatcg actgcacggt gcaccaatgc
7021 ttctggcgtc aggcagccat cgg
//
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