| 质粒名称: | pSLIK-Neo |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 25735 |
| 存储实验室: | Iain Fraser |
|
载体骨架基本信息 |
|
| 载体名称: | pSLIK |
| 载体出品公司: | Iain Fraser |
| 空载体大小: | 13286 bp |
| 完整质粒大小: | 13290bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | Chloramphenicol and Ampicillin, 25 & 100 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DB3.1 |
| 拷贝数: | 未知 |
|
插入基因信息 |
|
| 克隆方法: | Gateway Cloning |
| 5’克隆位点: | attR1(未破坏) |
| 3’克隆位点: | attR2 (未破坏) |
| 5’测序引物 | CMV-F |
| 3’测序引物 | hUBCpro-R (CTAAGGCCGAGTCTTATGAGCAG) |
|
文献引用方法 |
|
| 材料和方法部分: | pSLIK-Neo was a gift from Iain Fraser (Addgene plasmid # 25735 ; http://n2t.net/addgene:25735 ; RRID:Addgene_25735) |
| 参考文献部分: | A single lentiviral vector platform for microRNA-based conditional RNA interference and coordinated transgene expression. Shin KJ, Wall EA, Zavzavadjian JR, Santat LA, Liu J, Hwang JI, Rebres R, Roach T, Seaman W, Simon MI, Fraser ID. Proc Natl Acad Sci U S A. 2006 Sep 12. 103(37):13759-64. 10.1073/pnas.0606179103 PubMed 16945906 |
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 pSLIK-Neo 13290 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION 3rd generation Lentiviral vector for Tet-based inducible shRNA or
cDNA expression, constitutive Neomycin resistance gene
coexpression..
ACCESSION .
VERSION .
KEYWORDS pSLIK-Neo
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13290)
AUTHORS Shin KJ, Wall EA, Zavzavadjian JR, Santat LA, Liu J, Hwang JI,
Rebres R, Roach T, Seaman W, Simon MI, Fraser ID
TITLE A single lentiviral vector platform for microRNA-based conditional
RNA interference and coordinated transgene expression.
JOURNAL Proc Natl Acad Sci U S A. 2006 Sep 12. 103(37):13759-64.
PUBMED 16945906
REFERENCE 2 (bases 1 to 13290)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..13290
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 382..580
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 535..555
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 598..778
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 825..950
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1443..1676
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1861..1905
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
misc_feature 2203..2320
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
protein_bind 2377..2501
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 2526..2556
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 2610..3269
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(2676..2695)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
CDS 3611..3916
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
primer_bind 3795..3814
/label=ccdB-fwd
/note="ccdB gene, forward primer"
protein_bind complement(3957..4081)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 4131..5342
/label=UbC promoter
/note="human ubiquitin C promoter"
primer_bind 5179..5198
/label=hUBCpro-F
/note="Human Ubiquitin C (UbC) promoter, forward primer"
CDS 5401..6147
/codon_start=1
/product="improved tetracycline-controlled transactivator"
/label=rtTA-Advanced
/note="In the Tet-On(R) system, rtTA-Advanced binds to the
Tet-responsive element and stimulates transcription only in
the presence of tetracycline or doxycycline."
/translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
DALDDFDLDMLPADALDDFDLDMLPG"
misc_feature 6301..6763
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(6357..6374)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 6584..6603
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 6779..7573
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(6833..6852)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 7443..7462
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
misc_feature 7741..8329
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(7794..7814)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(8212..8223)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(8332..8348)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8333..8349)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 8858..9038
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
primer_bind complement(9064..9081)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 9070..9294
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 9340..9768
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(9427..9446)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 9637..9658
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(9777..9797)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 9782..10111
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 9962..10097
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 10024..10043
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 10159..10206
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 10225..10599
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=BleoR
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
polyA_signal 10729..10850
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(10766..10785)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 10820..10839
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(10899..10915)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(10899..10915)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(10912..10934)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 10923..10939
/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(10947..10977)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 10992..11013
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(11130..11147)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(11301..11889)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(11381..11400)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(12060..12920)
/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 12683..12702
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(12921..13025)
/gene="bla"
/label=AmpR promoter
primer_bind complement(13100..13119)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
2341 cagagatcca gtttggttaa ttctagacag ttatcaacaa gtttgtacaa aaaagctgaa
2401 cgagaaacgt aaaatgatat aaatatcaat atattaaatt agattttgca taaaaaacag
2461 actacataat actgtaaaac acaacatatc cagtcactat ggcggccgca ttaggcaccc
2521 caggctttac actttatgct tccggctcgt ataatgtgtg gattttgagt taggatccgt
2581 cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga tataccaccg
2641 ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat
2701 gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc gtaaagaaaa
2761 ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg aatgctcatc
2821 cggaattccg tatggcaatg aaagacggtg agctggtgat atgggatagt gttcaccctt
2881 gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt gaataccacg
2941 acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc
3001 tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc aatccctggg
3061 tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt
3121 tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg
3181 ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt
3241 actgcgatga gtggcagggc ggggcgtaaa gatctggatc cggcttacta aaagccagat
3301 aacagtatgc gtatttgcgc gctgattttt gcggtataag aatatatact gatatgtata
3361 cccgaagtat gtcaaaaaga ggtatgctat gaagcagcgt attacagtga cagttgacag
3421 cgacagctat cagttgctca aggcatatat gatgtcaata tctccggtct ggtaagcaca
3481 accatgcaga atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga aaatcaggaa
3541 gggatggctg aggtcgcccg gtttattgaa atgaacggct cttttgctga cgagaacagg
3601 ggctggtgaa atgcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt
3661 tgtggatgta cagagtgata ttattgacac gcccgggcga cggatggtga tccccctggc
3721 cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg
3781 ggatgaaagc tggcgcatga tgaccaccga tatggccagt gtgccggtct ccgttatcgg
3841 ggaagaagtg gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat
3901 gttctgggga atataaatgt caggctccct tatacacagc cagtctgcag gtcgaccata
3961 gtgactggat atgttgtgtt ttacagtatt atgtagtctg ttttttatgc aaaatctaat
4021 ttaatatatt gatatttata tcattttacg tttctcgttc agctttcttg tacaaagtgg
4081 ttgataacat ctcgagccat aaagatggtt aattaaccca cccaagatct ggcctccgcg
4141 ccgggttttg gcgcctcccg cgggcgcccc cctcctcacg gcgagcgctg ccacgtcaga
4201 cgaagggcgc agcgagcgtc ctgatccttc cgcccggacg ctcaggacag cggcccgctg
4261 ctcataagac tcggccttag aaccccagta tcagcagaag gacattttag gacgggactt
4321 gggtgactct agggcactgg ttttctttcc agagagcgga acaggcgagg aaaagtagtc
4381 ccttctcggc gattctgcgg agggatctcc gtggggcggt gaacgccgat gattatataa
4441 ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg ggatttgggt cgcggttctt
4501 gtttgtggat cgctgtgatc gtcacttggt gagtagcggg ctgctgggct ggccggggct
4561 ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc caagggctgt
4621 agtctgggtc cgcgagcaag gttgccctga actgggggtt ggggggagcg cagcaaaatg
4681 gcggctgttc ccgagtcttg aatggaagac gcttgtgagg cgggctgtga ggtcgttgaa
4741 acaaggtggg gggcatggtg ggcggcaaga acccaaggtc ttgaggcctt cgctaatgcg
4801 ggaaagctct tattcgggtg agatgggctg gggcaccatc tggggaccct gacgtgaagt
4861 ttgtcactga ctggagaact cggtttgtcg tctgttgcgg gggcggcagt tatggcggtg
4921 ccgttgggca gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc gtgacgtcac
4981 ccgttctgtt ggcttataat gcagggtggg gccacctgcc ggtaggtgtg cggtaggctt
5041 ttctccgtcg caggacgcag ggttcgggcc tagggtaggc tctcctgaat cgacaggcgc
5101 cggacctctg gtgaggggag ggataagtga ggcgtcagtt tctttggtcg gttttatgta
5161 cctatcttct taagtagctg aagctccggt tttgaactat gcgctcgggg ttggcgagtg
5221 tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt
5281 ttcagtgtta gactagtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag
5341 acgaagcttg gtaccgagct cggatctcca ccccgtaccg gtcctgcagt cgaattcacc
5401 atgtctagac tggacaagag caaagtcata aacggagctc tggaattact caatggtgtc
5461 ggtatcgaag gcctgacgac aaggaaactc gctcaaaagc tgggagttga gcagcctacc
5521 ctgtactggc acgtgaagaa caagcgggcc ctgctcgatg ccctgccaat cgagatgctg
5581 gacaggcatc atacccactt ctgccccctg gaaggcgagt catggcaaga ctttctgcgg
5641 aacaacgcca agtcataccg ctgtgctctc ctctcacatc gcgacggggc taaagtgcat
5701 ctcggcaccc gcccaacaga gaaacagtac gaaaccctgg aaaatcagct cgcgttcctg
5761 tgtcagcaag gcttctccct ggagaacgca ctgtacgctc tgtccgccgt gggccacttt
5821 acactgggct gcgtattgga ggaacaggag catcaagtag caaaagagga aagagagaca
5881 cctaccaccg attctatgcc cccacttctg agacaagcaa ttgagctgtt cgaccggcag
5941 ggagccgaac ctgccttcct tttcggcctg gaactaatca tatgtggcct ggagaaacag
6001 ctaaagtgcg aaagcggcgg gccgaccgac gcccttgacg attttgactt agacatgctc
6061 ccagccgatg cccttgacga ctttgacctt gatatgctgc ctgctgacgc tcttgacgat
6121 tttgaccttg acatgctccc cgggtaacta agtaaggatc cgcggccgca ctagaggaat
6181 tccgcccctc tccccccccc ccctctcccc taacgttact ggccgaagcc gcttggaata
6241 aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt
6301 gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct
6361 cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc
6421 ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga
6481 caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc
6541 ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt
6601 attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg
6661 gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc
6721 gaaccacggg gacgtggttt tcctttgaaa aacacgatga taagcttacc ggtccaccat
6781 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
6841 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
6901 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
6961 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
7021 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
7081 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
7141 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
7201 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
7261 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
7321 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
7381 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
7441 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
7501 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
7561 cgagttcttc tgatgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca
7621 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
7681 ataaaatgaa tgcaattgtt gttgtttagt ccctcccaat tcgatatcaa gcttatcgat
7741 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
7801 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
7861 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
7921 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
7981 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
8041 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
8101 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
8161 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
8221 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
8281 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccgt
8341 cgacctcgat cgagacctag aaaaacatgg agcaatcaca agtagcaata cagcagctac
8401 caatgctgat tgtgcctggc tagaagcaca agaggaggag gaggtgggtt ttccagtcac
8461 acctcaggta cctttaagac caatgactta caaggcagct gtagatctta gccacttttt
8521 aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag atatccttga
8581 tctgtggatc taccacacac aaggctactt ccctgattgg cagaactaca caccagggcc
8641 agggatcaga tatccactga cctttggatg gtgctacaag ctagtaccag ttgagcaaga
8701 gaaggtagaa gaagccaatg aaggagagaa cacccgcttg ttacaccctg tgagcctgca
8761 tgggatggat gacccggaga gagaagtatt agagtggagg tttgacagcc gcctagcatt
8821 tcatcacatg gcccgagagc tgcatccgga ctgtactggg tctctctggt tagaccagat
8881 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
8941 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
9001 cctcagaccc ttttagtcag tgtggaaaat ctctagcagg gcccgtttaa acccgctgat
9061 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
9121 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
9181 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
9241 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
9301 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
9361 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
9421 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
9481 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
9541 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
9601 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
9661 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
9721 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
9781 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
9841 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
9901 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
9961 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
10021 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
10081 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
10141 cggatctgat cagcacgtgt tgacaattaa tcatcggcat agtatatcgg catagtataa
10201 tacgacaagg tgaggaacta aaccatggcc aagttgacca gtgccgttcc ggtgctcacc
10261 gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac
10321 ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg
10381 gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac
10441 gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg
10501 gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc
10561 ggcaactgcg tgcacttcgt ggccgaggag caggactgac acgtgctacg agatttcgat
10621 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
10681 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
10741 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
10801 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
10861 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
10921 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
10981 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
11041 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
11101 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
11161 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
11221 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
11281 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
11341 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
11401 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
11461 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
11521 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
11581 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
11641 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
11701 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
11761 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
11821 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
11881 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
11941 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
12001 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
12061 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
12121 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
12181 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
12241 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
12301 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
12361 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
12421 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
12481 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
12541 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
12601 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
12661 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
12721 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
12781 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
12841 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
12901 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
12961 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
13021 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
13081 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
13141 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
13201 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
13261 ttcgcgatgt acgggccaga tatacgcgtt
//
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