| 质粒名称: | pET-28a_6H-TAQ_E602D |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 166944 |
| 存储实验室: | Robert Tjian |
|
载体骨架基本信息 |
|
| 载体名称: | pET-28a(+) |
| 载体出品公司: | Novagen (EMD Millipore) |
| 空载体大小: | 5369 bp |
| 完整质粒大小: | 7826 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 载体抗性: | 卡那霉素50 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | -- |
|
插入基因信息 |
|
| 插入基因名称: | Thermus aquaticus DNA polymerase,TAQ polymerase |
| 物种来源: | Thermus aquaticus |
| 插入大小: | 2499 bp |
| 基因突变: | E602D |
| GenBank编号 | J04639 |
| 启动子: | T7 |
| 标签/融合蛋白: | 6X His（主干上的N端子） |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | NdeI(未破坏) |
| 3’克隆位点: | EcoRI(未破坏) |
| 5’测序引物 | taatacgactcactataggg |
| 3’测序引物 | TAGTTATTGCTCAGCGGTGG |
|
文献引用方法 |
|
| 材料和方法部分: | pET-28a_6H-TAQ_E602D was a gift from Robert Tjian (Addgene plasmid # 166944 ; http://n2t.net/addgene:166944 ; RRID:Addgene_166944) |
| 参考文献部分: | Open-source RNA extraction and RT-qPCR methods for SARS-CoV-2 detection. Graham TGW, Dugast-Darzacq C, Dailey GM, Nguyenla XH, Van Dis E, Esbin MN, Abidi A, Stanley SA, Darzacq X, Tjian R. PLoS One. 2021 Feb 3;16(2):e0246647. doi: 10.1371/journal.pone.0246647. eCollection 2021. 10.1371/journal.pone.0246647 PubMed 33534838 |
买家导航
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 pET-28a_6H-TAQ_E602D 7822 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Bacterial expression of Taq DNA polymerase.
ACCESSION .
VERSION .
KEYWORDS pET-28a_6H-TAQ_E602D
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7822)
AUTHORS Graham TGW, Dugast-Darzacq C, Dailey GM, Nguyenla XH, Van Dis E,
Esbin MN, Abidi A, Stanley SA, Darzacq X, Tjian R
TITLE Open-source RNA extraction and RT-qPCR methods for SARS-CoV-2
detection.
JOURNAL PLoS One. 2021 Feb 3;16(2):e0246647. doi:
10.1371/journal.pone.0246647. eCollection 2021.
PUBMED 33534838
REFERENCE 2 (bases 1 to 7822)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7822
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 1..25
/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)."
RBS 40..62
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 81..98
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 108..125
/codon_start=1
/product="thrombin recognition and cleavage site"
/label=thrombin site
/translation="LVPRGS"
CDS 129..2627
/codon_start=1
/product="thermostable DNA polymerase from Thermus
aquaticus"
/label=Taq polymerase
/translation="MRGMLPLFEPKGRVLLVDGHHLAYRTFHALKGLTTSRGEPVQAVY
GFAKSLLKALKEDGDAVIVVFDAKAPSFRHEAYGGYKAGRAPTPEDFPRQLALIKELVD
LLGLARLEVPGYEADDVLASLAKKAEKEGYEVRILTADKDLYQLLSDRIHVLHPEGYLI
TPAWLWEKYGLRPDQWADYRALTGDESDNLPGVKGIGEKTARKLLEEWGSLEALLKNLD
RLKPAIREKILAHMDDLKLSWDLAKVRTDLPLEVDFAKRREPDRERLRAFLERLEFGSL
LHEFGLLESPKALEEAPWPPPEGAFVGFVLSRKEPMWADLLALAAARGGRVHRAPEPYK
ALRDLKEARGLLAKDLSVLALREGLGLPPGDDPMLLAYLLDPSNTTPEGVARRYGGEWT
EEAGERAALSERLFANLWGRLEGEERLLWLYREVERPLSAVLAHMEATGVRLDVAYLRA
LSLEVAEEIARLEAEVFRLAGHPFNLNSRDQLERVLFDELGLPAIGKTEKTGKRSTSAA
VLEALREAHPIVEKILQYRELTKLKSTYIDPLPDLIHPRTGRLHTRFNQTATATGRLSS
SDPNLQNIPVRTPLGQRIRRAFIAEDGWLLVALDYSQIELRVLAHLSGDENLIRVFQEG
RDIHTETASWMFGVPREAVDPLMRRAAKTINFGVLYGMSAHRLSQELAIPYEEAQAFIE
RYFQSFPKVRAWIEKTLEEGRRRGYVETLFGRRRYVPDLEARVKSVREAAERMAFNMPV
QGTAADLMKLAMVKLFPRLEEMGARMLLQVHDELVLEAPKERAEAVARLAKEVMEGVYP
LAVPLEVEVGIGEDWLSAKE"
CDS 2668..2685
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(2738..2756)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 2752..2799
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 2836..3291
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2923..2942)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3133..3154
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(3383..4198)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 4106..4125
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 4190..4209
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 4320..4908
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4809..4828
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5062..5079
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 5094..5233
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(5159..5178)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 5319..5341
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(5335..5526)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(6335..7417)
/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 7379..7398
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(7418..7495)
/gene="lacI"
/label=lacI promoter
primer_bind 7701..7720
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 7804..1
/label=T7
/note="T7 promoter, forward primer"
promoter 7804..7822
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga
61 gatataccat gggcagcagc catcatcatc atcatcacag cagcggcctg gtgccgcgcg
121 gcagccatat gagggggatg ctgcccctct ttgagcccaa gggccgggtc ctcctggtgg
181 acggccacca cctggcctac cgcaccttcc acgccctgaa gggcctcacc accagccggg
241 gggagccggt gcaggcggtc tacggcttcg ccaagagcct cctcaaggcc ctcaaggagg
301 acggggacgc ggtgatcgtg gtctttgacg ccaaggcccc ctccttccgc cacgaggcct
361 acggggggta caaggcgggc cgggccccca cgccggagga ctttccccgg caactcgccc
421 tcatcaagga gctggtggac ctcctggggc tggcgcgcct cgaggtcccg ggctacgagg
481 cggacgacgt cctggccagc ctggccaaga aggcggaaaa ggagggctac gaggtccgca
541 tcctcaccgc cgacaaagac ctttaccagc tcctttccga ccgcattcac gtcctccacc
601 ccgaggggta cctcatcacc ccggcctggc tttgggaaaa gtacggcctg aggcccgacc
661 agtgggccga ctaccgggcc ctgaccgggg acgagtccga caaccttccc ggggtcaagg
721 gcatcgggga gaagacggcg aggaagcttc tggaggagtg ggggagcctg gaagccctcc
781 tcaagaacct ggaccggctg aagcccgcca tccgggagaa gatcctggcc cacatggacg
841 atctgaagct ctcctgggac ctggccaagg tgcgcaccga cctgcccctg gaggtggact
901 tcgccaaaag gcgggagccc gaccgggaga ggcttagggc ctttctggag aggcttgagt
961 ttggcagcct cctccacgag ttcggccttc tggaaagccc caaggccctg gaggaggccc
1021 cctggccccc gccggaaggg gccttcgtgg gctttgtgct ttcccgcaag gagcccatgt
1081 gggccgatct tctggccctg gccgccgcca gggggggccg ggtccaccgg gcccccgagc
1141 cttataaagc cctcagggac ctgaaggagg cgcgggggct tctcgccaaa gacctgagcg
1201 ttctggccct gagggaaggc cttggcctcc cgcccggcga cgaccccatg ctcctcgcct
1261 acctcctgga cccttccaac accacccccg agggggtggc ccggcgctac ggcggggagt
1321 ggacggagga ggcgggggag cgggccgccc tttccgagag gctcttcgcc aacctgtggg
1381 ggaggcttga gggggaggag aggctccttt ggctttaccg ggaggtggag aggccccttt
1441 ccgctgtcct ggcccacatg gaggccacgg gggtgcgcct ggacgtggcc tatctcaggg
1501 ccttgtccct ggaggtggcc gaggagatcg cccgcctcga ggccgaggtc ttccgcctgg
1561 ccggccaccc cttcaacctc aactcccggg accagctgga aagggtcctc tttgacgagc
1621 tagggcttcc cgccatcggc aagacggaga agaccggcaa gcgctccacc agcgccgccg
1681 tcctggaggc cctccgcgag gcccacccca tcgtggagaa gatcctgcag taccgggagc
1741 tcaccaagct gaagagcacc tacattgacc ccttgccgga cctcatccac cccaggacgg
1801 gccgcctcca cacccgcttc aaccagacgg ccacggccac gggcaggcta agtagctccg
1861 atcccaacct ccagaacatc cccgtccgca ccccgcttgg gcagaggatc cgccgggcct
1921 tcatcgccga ggatgggtgg ctattggtgg ccctggacta tagccagata gagctcaggg
1981 tgctggccca cctctccggc gacgagaacc tgatccgggt cttccaggag gggcgggaca
2041 tccacacgga gaccgccagc tggatgttcg gcgtcccccg ggaggccgtg gaccccctga
2101 tgcgccgggc ggccaagacc atcaacttcg gggtcctcta cggcatgtcg gcccaccgcc
2161 tctcccagga gctagccatc ccttacgagg aggcccaggc cttcattgag cgctactttc
2221 agagcttccc caaggtgcgg gcctggattg agaagaccct ggaggagggc aggaggcggg
2281 ggtacgtgga gaccctcttc ggccgccgcc gctacgtgcc agacctagag gcccgggtga
2341 agagcgtgcg ggaggcggcc gagcgcatgg ccttcaacat gcccgtccag ggcaccgccg
2401 ccgacctcat gaagctggct atggtgaagc tcttccccag gctggaggaa atgggggcca
2461 ggatgctcct tcaggtccac gacgagctgg tcctcgaggc cccaaaagag agggcggagg
2521 ccgtggcccg gctggccaag gaggtcatgg agggggtgta tcccctggcc gtgcccctgg
2581 aggtggaggt ggggataggg gaggactggc tctccgccaa ggagtgagaa ttcgagctcc
2641 gtcgacaagc ttgcggccgc actcgagcac caccaccacc accactgaga tccggctgct
2701 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
2761 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
2821 ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
2881 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
2941 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
3001 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
3061 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
3121 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
3181 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
3241 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
3301 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
3361 gtatccgctc atgaattaat tcttagaaaa actcatcgag catcaaatga aactgcaatt
3421 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
3481 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
3541 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
3601 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt
3661 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
3721 aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag
3781 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
3841 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg
3901 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
3961 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
4021 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga
4081 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
4141 ccatgttgga atttaatcgc ggcctagagc aagacgtttc ccgttgaata tggctcataa
4201 caccccttgt attactgttt atgtaagcag acagttttat tgttcatgac caaaatccct
4261 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
4321 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
4381 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
4441 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
4501 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
4561 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
4621 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
4681 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
4741 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
4801 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
4861 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
4921 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
4981 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
5041 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
5101 cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta
5161 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
5221 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
5281 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
5341 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
5401 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
5461 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
5521 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
5581 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
5641 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
5701 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
5761 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
5821 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
5881 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
5941 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc
6001 catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa
6061 ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc
6121 gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac
6181 gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca
6241 ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta
6301 atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
6361 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
6421 tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca
6481 ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa
6541 aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt
6601 atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg
6661 cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca
6721 gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta
6781 tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg
6841 agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat
6901 gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct
6961 ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg
7021 catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat
7081 tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc
7141 tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca
7201 gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg
7261 ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt
7321 tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg
7381 catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct
7441 cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga
7501 tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg
7561 ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc
7621 ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg
7681 cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg
7741 gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga
7801 aattaatacg actcactata gg
//
