| 质粒名称: | pREDIA |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 24390 |
| 存储实验室: | Sheng Yang |
|
载体骨架基本信息 |
|
| 载体名称: | pKD46 |
| 载体出品公司: | Clontech |
| 空载体大小: | 6300bp |
| 完整质粒大小: | 9030 bp |
| 载体类型: | Bacterial Expression ; Red recombinase and I-SceI endonuclease |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 30 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
/ mitochondrial intron of Saccharomyces cerevisiae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: | pRha |
| 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 | 无 |
|
文献引用方法 |
|
| 材料和方法部分: | pREDIA was a gift from Sheng Yang (Addgene plasmid # 51625 ; http://n2t.net/addgene:51625 ; RRID:Addgene_51625) |
| 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 pREDIA 9030 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION E. coli genome modification .
ACCESSION .
VERSION .
KEYWORDS pREDIA
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9030)
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 9030)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biopfeng.com
FEATURES Location/Qualifiers
source 1..9030
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(11..889)
/codon_start=1
/gene="araC"
/product="L-arabinose regulatory protein"
/label=araC
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
promoter 916..1200
/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 1123..1142
/label=pBAD Forward
/note="For vectors with E. coli araBAD promoter, forward
primer"
RBS 1227..1235
/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 1244..1660
/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 1666..2451
/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 2448..3128
/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 3129..3373
/label=-lambda tL3 terminator
/note="transcription terminator tL3 from phage lambda"
CDS complement(3406..4254)
/codon_start=1
/gene="E. coli rhaR"
/product="transcriptional activator of rhaR and rhaS"
/label=rhaR
/note="needed for rhamnose-dependent activation of the rhaB
promoter"
/translation="MAHQLKLLKDDFFASDQQAVAVADRYPQDVFAEHTHDFCELVIVW
RGNGLHVLNDRPYRITRGDLFYIHADDKHSYASVNDLVLQNIIYCPERLKLNLDWQGAI
PGFNASAGQPHWRLGSMGMAQARQVIGQLEHESSQHVPFANEMAELLFGQLVMLLNRHR
YTSDSLPPTSSETLLDKLITRLAASLKSPFALDKFCDEASCSERVLRQQFRQQTGMTIN
QYLRQVRVCHAQYLLQHSRLLISDISTECGFEDSNYFSVVFTRETGMTPSQWRHLNSQK
D"
CDS complement(4328..5164)
/codon_start=1
/gene="E. coli rhaS"
/product="positive regulator of the rhaB promoter
"
/label=rhaS
/note="activated by L-rhamnose"
/translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
promoter 5310..5428
/label=rhaB promoter
/note="promoter of the E. coli rhaBAD operon, conferring
tight induction with L-rhamnose and repression with
D-glucose in the presence of RhaR and RhaS (Giacalone et
al., 2006)"
CDS complement(6197..7147)
/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 7195..7417
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
CDS complement(8039..8899)
/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 8662..8681
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8900..9004)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 catcgattta ttatgacaac ttgacggcta catcattcac tttttcttca caaccggcac
61 ggaactcgct cgggctggcc ccggtgcatt ttttaaatac ccgcgagaaa tagagttgat
121 cgtcaaaacc aacattgcga ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca
181 gcttcgcctg gctgatacgt tggtcctcgc gccagcttaa gacgctaatc cctaactgct
241 ggcggaaaag atgtgacaga cgcgacggcg acaagcaaac atgctgtgcg acgctggcga
301 tatcaaaatt gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg cgtacccgat
361 tatccatcgg tggatggagc gactcgttaa tcgcttccat gcgccgcagt aacaattgct
421 caagcagatt tatcgccagc agctccgaat agcgcccttc cccttgcccg gcgttaatga
481 tttgcccaaa caggtcgctg aaatgcggct ggtgcgcttc atccgggcga aagaaccccg
541 tattggcaaa tattgacggc cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt
601 aaacccactg gtgataccat tcgcgagcct ccggatgacg accgtagtga tgaatctctc
661 ctggcgggaa cagcaaaata tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca
721 ccaccccctg accgcgaatg gtgagattga gaatataacc tttcattccc agcggtcggt
781 cgataaaaaa atcgagataa ccgttggcct caatcggcgt taaacccgcc accagatggg
841 cattaaacga gtatcccggc agcaggggat cattttgcgc ttcagccata cttttcatac
901 tcccgccatt cagagaagaa accaattgtc catattgcat cagacattgc cgtcactgcg
961 tcttttactg gctcttctcg ctaaccaaac cggtaacccc gcttattaaa agcattctgt
1021 aacaaagcgg gaccaaagcc atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca
1081 gaaaagtcca cattgattat ttgcacggcg tcacactttg ctatgccata gcatttttat
1141 ccataagatt agcggatcct acctgacgct ttttatcgca actctctact gtttctccat
1201 acccgttttt ttgggaattc gagctctaag gaggttataa aaaatggata ttaatactga
1261 aactgagatc aagcaaaagc attcactaac cccctttcct gttttcctaa tcagcccggc
1321 atttcgcggg cgatattttc acagctattt caggagttca gccatgaacg cttattacat
1381 tcaggatcgt cttgaggctc agagctgggc gcgtcactac cagcagctcg cccgtgaaga
1441 gaaagaggca gaactggcag acgacatgga aaaaggcctg ccccagcacc tgtttgaatc
1501 gctatgcatc gatcatttgc aacgccacgg ggccagcaaa aaatccatta cccgtgcgtt
1561 tgatgacgat gttgagtttc aggagcgcat ggcagaacac atccggtaca tggttgaaac
1621 cattgctcac caccaggttg atattgattc agaggtataa aacgaatgag tactgcactc
1681 gcaacgctgg ctgggaagct ggctgaacgt gtcggcatgg attctgtcga cccacaggaa
1741 ctgatcacca ctcttcgcca gacggcattt aaaggtgatg ccagcgatgc gcagttcatc
1801 gcattactga tcgttgccaa ccagtacggc cttaatccgt ggacgaaaga aatttacgcc
1861 tttcctgata agcagaatgg catcgttccg gtggtgggcg ttgatggctg gtcccgcatc
1921 atcaatgaaa accagcagtt tgatggcatg gactttgagc aggacaatga atcctgtaca
1981 tgccggattt accgcaagga ccgtaatcat ccgatctgcg ttaccgaatg gatggatgaa
2041 tgccgccgcg aaccattcaa aactcgcgaa ggcagagaaa tcacggggcc gtggcagtcg
2101 catcccaaac ggatgttacg tcataaagcc atgattcagt gtgcccgtct ggccttcgga
2161 tttgctggta tctatgacaa ggatgaagcc gagcgcattg tcgaaaatac tgcatacact
2221 gcagaacgtc agccggaacg cgacatcact ccggttaacg atgaaaccat gcaggagatt
2281 aacactctgc tgatcgccct ggataaaaca tgggatgacg acttattgcc gctctgttcc
2341 cagatatttc gccgcgacat tcgtgcatcg tcagaactga cacaggccga agcagtaaaa
2401 gctcttggat tcctgaaaca gaaagccgca gagcagaagg tggcagcatg acaccggaca
2461 ttatcctgca gcgtaccggg atcgatgtga gagctgtcga acagggggat gatgcgtggc
2521 acaaattacg gctcggcgtc atcaccgctt cagaagttca caacgtgata gcaaaacccc
2581 gctccggaaa gaagtggcct gacatgaaaa tgtcctactt ccacaccctg cttgctgagg
2641 tttgcaccgg tgtggctccg gaagttaacg ctaaagcact ggcctgggga aaacagtacg
2701 agaacgacgc cagaaccctg tttgaattca cttccggcgt gaatgttact gaatccccga
2761 tcatctatcg cgacgaaagt atgcgtaccg cctgctctcc cgatggttta tgcagtgacg
2821 gcaacggcct tgaactgaaa tgcccgttta cctcccggga tttcatgaag ttccggctcg
2881 gtggtttcga ggccataaag tcagcttaca tggcccaggt gcagtacagc atgtgggtga
2941 cgcgaaaaaa tgcctggtac tttgccaact atgacccgcg tatgaagcgt gaaggcctgc
3001 attatgtcgt gattgagcgg gatgaaaagt acatggcgag ttttgacgag atcgtgccgg
3061 agttcatcga aaaaatggac gaggcactgg ctgaaattgg ttttgtattt ggggagcaat
3121 ggcgatgacg catcctcacg ataatatccg ggtaggcgca atcactttcg tctactccgt
3181 tacaaagcga ggctgggtat ttcccggcct ttctgttatc cgaaatccac tgaaagcaca
3241 gcggctggct gaggagataa ataataaacg aggggctgta tgcacaaagc atcttctgtt
3301 gagttaagaa cgagtatcga gatggcacat agccttgctc aaattggaat caggtttgtg
3361 ccaataccag tagaaacaga cgaagaatcc atggggcatg gcgaattaat ctttctgcga
3421 attgagatga cgccactggc tgggcgtcat cccggtttcc cgggtaaaca ccaccgaaaa
3481 atagttacta tcttcaaagc cacattcggt cgaaatatca ctgattaaca ggcggctatg
3541 ctggagaaga tattgcgcat gacacactct gacctgtcgc agatattgat tgatggtcat
3601 tccagtctgc tggcgaaatt gctgacgcaa aacgcgctca ctgcacgatg cctcatcaca
3661 aaatttatcc agcgcaaagg gacttttcag gctagccgcc agccgggtaa tcagcttatc
3721 cagcaacgtt tcgctggatg ttggcggcaa cgaatcactg gtgtaacgat ggcgattcag
3781 caacatcacc aactgcccga acagcaactc agccatttcg ttagcaaacg gcacatgctg
3841 actactttca tgctcaagct gaccgataac ctgccgcgcc tgcgccatcc ccatgctacc
3901 taagcgccag tgtggttgcc ctgcgctggc gttaaatccc ggaatcgccc cctgccagtc
3961 aagattcagc ttcagacgct ccgggcaata aataatattc tgcaaaacca gatcgttaac
4021 ggaagcgtag gagtgtttat cgtcagcatg aatgtaaaag agatcgccac gggtaatgcg
4081 ataagggcga tcgttgagta catgcaggcc attaccgcgc cagacaatca ccagctcaca
4141 aaaatcatgt gtatgttcag caaagacatc ttgcggataa cggtcagcca cagcgactgc
4201 ctgctggtcg ctggcaaaaa aatcatcttt gagaagtttt aactgatgcg ccaccgtggc
4261 tacctcggcc agagaacgaa gttgattatt cgcaatatgg cgtacaaata cgttgagaag
4321 attcgcgtta ttgcagaaag ccatcccgtc cctggcgaat atcacgcggt gaccagttaa
4381 actctcggcg aaaaagcgtc gaaaagtggt tactgtcgct gaatccacag cgataggcga
4441 tgtcagtaac gctggcctcg ctgtggcgta gcagatgtcg ggctttcatc agtcgcaggc
4501 ggttcaggta tcgctgaggc gtcagtcccg tttgctgctt aagctgccga tgtagcgtac
4561 gcagtgaaag agaaaattga tccgccacgg catcccaatt cacctcatcg gcaaaatggt
4621 cctccagcca ggccagaagc aagttgagac gtgatgcgct gttttccagg ttctcctgca
4681 aactgctttt acgcagcaag agcagtaatt gcataaacaa gatctcgcga ctggcggtcg
4741 agggtaaatc attttcccct tcctgctgtt ccatctgtgc aaccagctgt cgcacctgct
4801 gcaatacgct gtggttaacg cgccagtgag acggatactg cccatccagc tcttgtggca
4861 gcaactgatt cagcccggcg agaaactgaa atcgatccgg cgagcgatac agcacattgg
4921 tcagacacag attatcggta tgttcataca gatgccgatc atgatcgcgt acgaaacaga
4981 ccgtgccacc ggtgatggta tagggctgcc cattaaacac atgaataccc gtgccatgtt
5041 cgacaatcac aatttcatga aaatcatgat gatgttcagg aaaatccgcc tgcgggagcc
5101 ggggttctat cgccacggac gcgttaccag acggaaaaaa atccacacta tgtaatacgg
5161 tcatactggc ctcctgatgt cgtcaacacg gcgaaatagt aatcacgagg tcaggttctt
5221 accttaaatt ttcgacggaa aaccacgtaa aaaacgtcga tttttcaaga tacagcgtga
5281 attttcagga aatgcggtga gcatcacatc accacaattc agcaaattgt gaacatcatc
5341 acgttcatct ttccctggtt gccaatggcc cattttcctg tcagtaacga gaaggtcgcg
5401 aattcaggcg ctttttagac tggtcgtaat gaaattcagc aggatcacat atgcatcaaa
5461 aaaaccaggt aatgaacctg ggtccgaact ctaaactgct gaaagaatac aaatcccagc
5521 tgatcgaact gaacatcgaa cagttcgaag caggtatcgg tctgatcctg ggtgatgctt
5581 acatccgttc tcgtgatgaa ggtaaaacct actgtatgca gttcgagtgg aaaaacaaag
5641 catacatgga ccacgtatgt ctgctgtacg atcagtgggt actgtctttc aaacaccaag
5701 ctttcaacaa actggctaac ctgttcatcg ttaacaacaa aaaaaccatc ccgaacaacc
5761 tggttgaaaa ctacctgacc ccgatgtctc tggcatactg gttcatggat gatggtggta
5821 aatgggatta caacaaaaac tctaccaaca aatcgatcgt actgaacacc cagtctttca
5881 ctttcgaaga agtagaatac ctggttaagg gtctgcgtaa caaattccaa ctgaactgtt
5941 acgtaaaaat caacaaaaac aaaccgatca tctacatcga ttctatgtct tacctgatct
6001 tctacaacct gatcaaaccg tacctgatcc cgcagatgat gtacaaactg ccgaacacta
6061 tctcctccga aactttcctg aaataataac catgggtatg gacagttttc cctttgatat
6121 gtaacggtga acagttgttc tacttttgtt tgttagtctt gatgcttcac tgatagatac
6181 aagagccata agaacctcag atccttccgt atttagccag tatgttctct agtgtggttc
6241 gttgtttttg cgtgagccat gagaacgaac cattgagatc atacttactt tgcatgtcac
6301 tcaaaaattt tgcctcaaaa ctggtgagct gaatttttgc agttaaagca tcgtgtagtg
6361 tttttcttag tccgttacgt aggtaggaat ctgatgtaat ggttgttggt attttgtcac
6421 cattcatttt tatctggttg ttctcaagtt cggttacgag atccatttgt ctatctagtt
6481 caacttggaa aatcaacgta tcagtcgggc ggcctcgctt atcaaccacc aatttcatat
6541 tgctgtaagt gtttaaatct ttacttattg gtttcaaaac ccattggtta agccttttaa
6601 actcatggta gttattttca agcattaaca tgaacttaaa ttcatcaagg ctaatctcta
6661 tatttgcctt gtgagttttc ttttgtgtta gttcttttaa taaccactca taaatcctca
6721 tagagtattt gttttcaaaa gacttaacat gttccagatt atattttatg aattttttta
6781 actggaaaag ataaggcaat atctcttcac taaaaactaa ttctaatttt tcgcttgaga
6841 acttggcata gtttgtccac tggaaaatct caaagccttt aaccaaagga ttcctgattt
6901 ccacagttct cgtcatcagc tctctggttg ctttagctaa tacaccataa gcattttccc
6961 tactgatgtt catcatctga gcgtattggt tataagtgaa cgataccgtc cgttctttcc
7021 ttgtagggtt ttcaatcgtg gggttgagta gtgccacaca gcataaaatt agcttggttt
7081 catgctccgt taagtcatag cgactaatcg ctagttcatt tgctttgaaa acaactaatt
7141 cagacataca tctcaattgg tctaggtgat tttaatcact ataccaattg agatgggcta
7201 gtcaatgata attactagtc cttttccttt gagttgtggg tatctgtaaa ttctgctaga
7261 cctttgctgg aaaacttgta aattctgcta gaccctctgt aaattccgct agacctttgt
7321 gtgttttttt tgtttatatt caagtggtta taatttatag aataaagaaa gaataaaaaa
7381 agataaaaag aatagatccc agccctgtgt ataactcact actttagtca gttccgcagt
7441 attacaaaag gatgtcgcaa acgctgtttg ctcctctaca aaacagacct taaaacccta
7501 aaggcttaag tagcaccctc gcaagctcgg ttgcggccgc aatcgggcaa atcgctgaat
7561 attccttttg tctccgacca tcaggcacct gagtcgctgt ctttttcgtg acattcagtt
7621 cgctgcgctc acggctctgg cagtgaatgg gggtaaatgg cactacaggc gccttttatg
7681 gattcatgca aggaaactac ccataataca agaaaagccc gtcacgggct tctcagggcg
7741 ttttatggcg ggtctgctat gtggtgctat ctgacttttt gctgttcagc agttcctgcc
7801 ctctgatttt ccagtctgac cacttcggat tatcccgtga caggtcattc agactggcta
7861 atgcacccag taaggcagcg gtatcatcaa cggggtctga cgctcagtgg aacgaaaact
7921 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
7981 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
8041 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
8101 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
8161 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
8221 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
8281 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
8341 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
8401 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
8461 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
8521 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
8581 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
8641 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
8701 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
8761 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
8821 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
8881 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
8941 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
9001 cgcgcacatt tccccgaaaa gtgccacctg
//
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