| 质粒名称: | pREDTKI |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 51628 |
| 存储实验室: | Sheng Yang |
|
载体骨架基本信息 |
|
| 载体名称: | pKD46 |
| 空载体大小: | 6300bp |
| 完整质粒大小: | 8464 bp |
| 载体类型: | Bacterial Expression ; Red recombinase and I-SceI endonuclease |
| 载体抗性: | Kanamycin |
| 生长温度: | 30 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | I-SceI / mitochondrial intron of Saccharomyces cerevisiae |
| 物种来源: | S. cerevisiae (budding yeast) |
| 插入大小: | 3000 bp |
| 启动子: | trc |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | NcoI(未破坏) |
| 3’克隆位点: | NcoI(未破坏) |
| 5’测序引物 | none |
|
文献引用方法 |
|
| 材料和方法部分: | pREDTKI was a gift from Sheng Yang (Addgene plasmid # 51628 ; http://n2t.net/addgene:51628 ; RRID:Addgene_51628) |
| 参考文献部分: | High-Efficiency Scarless Genetic Modification in Escherichia coli by Using Lambda Red Recombination and I-SceI Cleavage. Yang J, Sun B, Huang H, Jiang Y, Diao L, Chen B, Xu C, Wang X, Liu J, Jiang W, Yang S. Appl Environ Microbiol. 2014 Jul 1;80(13):3826-3834. Epub 2014 Apr 18. 10.1128/AEM.00313-14 PubMed 24747889 |
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 pREDTKI 8464 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION E coli Genetic Modification.
ACCESSION .
VERSION .
KEYWORDS pREDTKI
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8464)
AUTHORS Yang J, Sun B, Huang H, Jiang Y, Diao L, Chen B, Xu C, Wang X, Liu
J, Jiang W, Yang S
TITLE High-Efficiency Scarless Genetic Modification in Escherichia coli by
Using Lambda Red Recombination and I-SceI Cleavage.
JOURNAL Appl Environ Microbiol. 2014 Jul 1;80(13):3826-3834. Epub 2014 Apr
18.
PUBMED 24747889
REFERENCE 2 (bases 1 to 8464)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8464
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(19..897)
/codon_start=1
/gene="araC"
/product="L-arabinose regulatory protein"
/label=araC
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
promoter 924..1208
/gene="araBAD"
/label=araBAD promoter
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
primer_bind 1131..1150
/label=pBAD Forward
/note="For vectors with E. coli araBAD promoter, forward
primer"
RBS 1235..1243
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
CDS 1252..1668
/codon_start=1
/gene="gam"
/product="inhibitor of the host RecBCD nuclease in the
lambda Red system"
/label=Gam
/note="protects the double-stranded DNA substrate for
recombination"
/translation="MDINTETEIKQKHSLTPFPVFLISPAFRGRYFHSYFRSSAMNAYY
IQDRLEAQSWARHYQQLAREEKEAELADDMEKGLPQHLFESLCIDHLQRHGASKKSITR
AFDDDVEFQERMAEHIRYMVETIAHHQVDIDSEV"
CDS 1674..2459
/codon_start=1
/gene="bet"
/product="single-stranded DNA binding recombinase in the
lambda Red system"
/label=Beta
/note="mediates pairing of a single-stranded DNA with a
complementary target"
/translation="MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFI
ALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESC
TCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLA
FGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLP
LCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA"
CDS 2456..3136
/codon_start=1
/gene="exo"
/product="5' to 3' double-stranded DNA exonuclease in the
lambda Red system"
/label=Exo
/note="creates 3' single-stranded DNA tails"
/translation="MTPDIILQRTGIDVRAVEQGDDAWHKLRLGVITASEVHNVIAKPR
SGKKWPDMKMSYFHTLLAEVCTGVAPEVNAKALAWGKQYENDARTLFEFTSGVNVTESP
IIYRDESMRTACSPDGLCSDGNGLELKCPFTSRDFMKFRLGGFEAIKSAYMAQVQYSMW
VTRKNAWYFANYDPRMKREGLHYVVIERDEKYMASFDEIVPEFIEKMDEALAEIGFVFG
EQWR"
terminator 3137..3381
/label=-lambda tL3 terminator
/note="transcription terminator tL3 from phage lambda"
primer_bind complement(4051..4073)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4062..4078
/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)."
promoter complement(4086..4115)
/label=trc promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
CDS complement(4347..5429)
/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
YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind 5391..5410
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(5430..5507)
/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 complement(5641..6591)
/codon_start=1
/gene="rep101"
/product="RepA protein needed for replication with the
pSC101 origin"
/label=Rep101
/translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
rep_origin 6639..6861
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
primer_bind complement(7616..7635)
/label=pENTR-R
/note="pENTR vectors, reverse primer"
CDS 7627..8442
/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 complement(7700..7719)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
ORIGIN
1 aaatggttta aacatttatt atgacaactt gacggctaca tcattcactt tttcttcaca
61 accggcacgg aactcgctcg ggctggcccc ggtgcatttt ttaaataccc gcgagaaata
121 gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc gggtggtgct
181 caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga cgctaatccc
241 taactgctgg cggaaaagat gtgacagacg cgacggcgac aagcaaacat gctgtgcgac
301 gctggcgata tcaaaattgc tgtctgccag gtgatcgctg atgtactgac aagcctcgcg
361 tacccgatta tccatcggtg gatggagcga ctcgttaatc gcttccatgc gccgcagtaa
421 caattgctca agcagattta tcgccagcag ctccgaatag cgcccttccc cttgcccggc
481 gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat ccgggcgaaa
541 gaaccccgta ttggcaaata ttgacggcca gttaagccat tcatgccagt aggcgcgcgg
601 acgaaagtaa acccactggt gataccattc gcgagcctcc ggatgacgac cgtagtgatg
661 aatctctcct ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt ctcgtccctg
721 atttttcacc accccctgac cgcgaatggt gagattgaga atataacctt tcattcccag
781 cggtcggtcg ataaaaaaat cgagataacc gttggcctca atcggcgtta aacccgccac
841 cagatgggca ttaaacgagt atcccggcag caggggatca ttttgcgctt cagccatact
901 tttcatactc ccgccattca gagaagaaac caattgtcca tattgcatca gacattgccg
961 tcactgcgtc ttttactggc tcttctcgct aaccaaaccg gtaaccccgc ttattaaaag
1021 cattctgtaa caaagcggga ccaaagccat gacaaaaacg cgtaacaaaa gtgtctataa
1081 tcacggcaga aaagtccaca ttgattattt gcacggcgtc acactttgct atgccatagc
1141 atttttatcc ataagattag cggatcctac ctgacgcttt ttatcgcaac tctctactgt
1201 ttctccatac ccgttttttt gggaattcga gctctaagga ggttataaaa aatggatatt
1261 aatactgaaa ctgagatcaa gcaaaagcat tcactaaccc cctttcctgt tttcctaatc
1321 agcccggcat ttcgcgggcg atattttcac agctatttca ggagttcagc catgaacgct
1381 tattacattc aggatcgtct tgaggctcag agctgggcgc gtcactacca gcagctcgcc
1441 cgtgaagaga aagaggcaga actggcagac gacatggaaa aaggcctgcc ccagcacctg
1501 tttgaatcgc tatgcatcga tcatttgcaa cgccacgggg ccagcaaaaa atccattacc
1561 cgtgcgtttg atgacgatgt tgagtttcag gagcgcatgg cagaacacat ccggtacatg
1621 gttgaaacca ttgctcacca ccaggttgat attgattcag aggtataaaa cgaatgagta
1681 ctgcactcgc aacgctggct gggaagctgg ctgaacgtgt cggcatggat tctgtcgacc
1741 cacaggaact gatcaccact cttcgccaga cggcatttaa aggtgatgcc agcgatgcgc
1801 agttcatcgc attactgatc gttgccaacc agtacggcct taatccgtgg acgaaagaaa
1861 tttacgcctt tcctgataag cagaatggca tcgttccggt ggtgggcgtt gatggctggt
1921 cccgcatcat caatgaaaac cagcagtttg atggcatgga ctttgagcag gacaatgaat
1981 cctgtacatg ccggatttac cgcaaggacc gtaatcatcc gatctgcgtt accgaatgga
2041 tggatgaatg ccgccgcgaa ccattcaaaa ctcgcgaagg cagagaaatc acggggccgt
2101 ggcagtcgca tcccaaacgg atgttacgtc ataaagccat gattcagtgt gcccgtctgg
2161 ccttcggatt tgctggtatc tatgacaagg atgaagccga gcgcattgtc gaaaatactg
2221 catacactgc agaacgtcag ccggaacgcg acatcactcc ggttaacgat gaaaccatgc
2281 aggagattaa cactctgctg atcgccctgg ataaaacatg ggatgacgac ttattgccgc
2341 tctgttccca gatatttcgc cgcgacattc gtgcatcgtc agaactgaca caggccgaag
2401 cagtaaaagc tcttggattc ctgaaacaga aagccgcaga gcagaaggtg gcagcatgac
2461 accggacatt atcctgcagc gtaccgggat cgatgtgaga gctgtcgaac agggggatga
2521 tgcgtggcac aaattacggc tcggcgtcat caccgcttca gaagttcaca acgtgatagc
2581 aaaaccccgc tccggaaaga agtggcctga catgaaaatg tcctacttcc acaccctgct
2641 tgctgaggtt tgcaccggtg tggctccgga agttaacgct aaagcactgg cctggggaaa
2701 acagtacgag aacgacgcca gaaccctgtt tgaattcact tccggcgtga atgttactga
2761 atccccgatc atctatcgcg acgaaagtat gcgtaccgcc tgctctcccg atggtttatg
2821 cagtgacggc aacggccttg aactgaaatg cccgtttacc tcccgggatt tcatgaagtt
2881 ccggctcggt ggtttcgagg ccataaagtc agcttacatg gcccaggtgc agtacagcat
2941 gtgggtgacg cgaaaaaatg cctggtactt tgccaactat gacccgcgta tgaagcgtga
3001 aggcctgcat tatgtcgtga ttgagcggga tgaaaagtac atggcgagtt ttgacgagat
3061 cgtgccggag ttcatcgaaa aaatggacga ggcactggct gaaattggtt ttgtatttgg
3121 ggagcaatgg cgatgacgca tcctcacgat aatatccggg taggcgcaat cactttcgtc
3181 tactccgtta caaagcgagg ctgggtattt cccggccttt ctgttatccg aaatccactg
3241 aaagcacagc ggctggctga ggagataaat aataaacgag gggctgtatg cacaaagcat
3301 cttctgttga gttaagaacg agtatcgaga tggcacatag ccttgctcaa attggaatca
3361 ggtttgtgcc aataccagta gaaacagacg aagaatccat ggttattatt tcaggaaagt
3421 ttcggaggag atagtgttcg gcagtttgta catcatctgc gggatcaggt acggtttgat
3481 caggttgtag aagatcaggt aagacataga atcgatgtag atgatcggtt tgtttttgtt
3541 gatttttacg taacagttca gttggaattt gttacgcaga cccttaacca ggtattctac
3601 ttcttcgaaa gtgaaagact gggtgttcag tacgatcgat ttgttggtag agtttttgtt
3661 gtaatcccat ttaccaccat catccatgaa ccagtatgcc agagacatcg gggtcaggta
3721 gttttcaacc aggttgttcg ggatggtttt tttgttgtta acgatgaaca ggttagccag
3781 tttgttgaaa gcttggtgtt tgaaagacag tacccactga tcgtacagca gacatacgtg
3841 gtccatgtat gctttgtttt tccactcgaa ctgcatacag taggttttac cttcatcacg
3901 agaacggatg taagcatcac ccaggatcag accgatacct gcttcgaact gttcgatgtt
3961 cagttcgatc agctgggatt tgtattcttt cagcagttta gagttcggac ccaggttcat
4021 tacctggttt ttttgatgca tatgctgttt cctgtgtgaa attgttatcc gctcacaatt
4081 ccacacatta tacgagccgg atgattaatt gtcaacagct catttcagaa tatttgccag
4141 aaccgttatg atgtcggcgc aaaaaacatt atccagaacg ggagtgcgcc ttgagcgaca
4201 cgaattatgc agtgatttac gacctgcaca gccataccac agcttccgat ggctgcctga
4261 cgccagaagc attggtgcac cgtgcagtcg atgataagct gtcaaaccag atcaattcgc
4321 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
4381 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc
4441 agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg
4501 ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt
4561 ttgatggtgg ttgacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact
4621 accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc
4681 gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc
4741 atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga
4801 atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa
4861 cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg
4921 cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag
4981 acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg
5041 tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc
5101 gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc
5161 agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga
5221 ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg
5281 ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa
5341 acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct
5401 gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg
5461 cgctatcatg ccataccgcg aaaggttttg caccattcga tggtgtcaac gtaaatgcat
5521 gccgcttcgc cttccatggg tatggacagt tttccctttg atatgtaacg gtgaacagtt
5581 gttctacttt tgtttgttag tcttgatgct tcactgatag atacaagagc cataagaacc
5641 tcagatcctt ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag
5701 ccatgagaac gaaccattga gatcatactt actttgcatg tcactcaaaa attttgcctc
5761 aaaactggtg agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt
5821 acgtaggtag gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg
5881 gttgttctca agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa
5941 cgtatcagtc gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa
6001 atctttactt attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt
6061 ttcaagcatt aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt
6121 tttcttttgt gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc
6181 aaaagactta acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg
6241 caatatctct tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt
6301 ccactggaaa atctcaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat
6361 cagctctctg gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat
6421 ctgagcgtat tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat
6481 cgtggggttg agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc
6541 atagcgacta atcgctagtt catttgcttt gaaaacaact aattcagaca tacatctcaa
6601 ttggtctagg tgattttaat cactatacca attgagatgg gctagtcaat gataattact
6661 agtccttttc ctttgagttg tgggtatctg taaattctgc tagacctttg ctggaaaact
6721 tgtaaattct gctagaccct ctgtaaattc cgctagacct ttgtgtgttt tttttgttta
6781 tattcaagtg gttataattt atagaataaa gaaagaataa aaaaagataa aaagaataga
6841 tcccagccct gtgtataact cactacttta gtcagttccg cagtattaca aaaggatgtc
6901 gcaaacgctg tttgctcctc tacaaaacag accttaaaac cctaaaggct taagtagcac
6961 cctcgcaagc tcggttgcgg ccgcaatcgg gcaaatcgct gaatattcct tttgtctccg
7021 accatcaggc acctgagtcg ctgtcttttt cgtgacattc agttcgctgc gctcacggct
7081 ctggcagtga atgggggtaa atggcactac aggcgccttt tatggattca tgcaaggaaa
7141 ctacccataa tacaagaaaa gcccgtcacg ggcttctcag ggcgttttat ggcgggtctg
7201 ctatgtggtg ctatctgact ttttgctgtt cagcagttcc tgccctctga ttttccagtc
7261 tgaccacttc ggattatccc gtgacaggtc attcagactg gctaatgcac ccagtaaggc
7321 agcggtatca tcaacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
7381 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
7441 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttacgttt aaaccatttc
7501 ctgcaggggg ggggggaaag ccacgttgtg tctcaaaatc tctgatgtta cattgcacaa
7561 gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
7621 ggtgttatga gccatattca acgggaaacg tcttgctcga ggccgcgatt aaattccaac
7681 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
7741 acaatctatc gattgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
7801 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
7861 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
7921 actgcgatcc ccgggaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
7981 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
8041 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac
8101 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
8161 tggaaagaaa tgcataagct tttgccattc tcaccggatt cagtcgtcac tcatggtgat
8221 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
8281 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
8341 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
8401 aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg
8461 ttgg
//
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