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addgene-plasmid-21916-sequence-358974 (TetR plasmid).gbk
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addgene-plasmid-21916-sequence-358974 (TetR plasmid).gbk
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LOCUS Exported 10959 bp ds-DNA circular SYN 12-APR-2022
DEFINITION 3rd generation lentiviral plasmid for inducible expression of shRNA;
neomycin selection. See manual for detailed protocols. No further
technical support is available from the depositing laboratory..
ACCESSION .
VERSION .
KEYWORDS Tet-pLKO-neo
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10959)
AUTHORS Wiederschain D, Wee S, Chen L, Loo A, Yang G, Huang A, Chen Y,
Caponigro G, Yao YM, Lengauer C, Sellers WR, Benson JD
TITLE Single-vector inducible lentiviral RNAi system for oncology target
validation.
JOURNAL Cell Cycle. 2009 Feb 1. 8(3):498-504.
PUBMED 19177017
REFERENCE 2 (bases 1 to 10959)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 12, 2022 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..10959
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 420..438
/label=hPGK-F
/note="Human PGK promoter, forward primer"
intron 510..1082
/label=-beta-globin intron
/note="intron from rabbit beta-globin gene"
primer_bind 1090..1109
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
promoter 1137..1155
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1166..1783
/codon_start=1
/gene="tetR from transposon Tn10"
/product="tetracycline repressor TetR"
/label=TetR
/note="TetR binds to the tetracycline operator tetO to
inhibit transcription. This inhibition can be relieved by
adding tetracycline or doxycycline."
/translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESG"
primer_bind complement(1239..1258)
/label=Tet-R
/note="Tetracycline resistance gene, reverse primer"
misc_feature 1821..2394
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(1988..2005)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 2215..2234
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
CDS 2401..3195
/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
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(2455..2474)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3065..3084
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
protein_bind complement(3224..3323)
/gene="mutant version of attL"
/label=attL2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
LTR 3467..3700
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 3772..3893
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3809..3828)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3863..3882
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 3933..4068
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3995..4014
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter complement(4089..4107)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4117..4134)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(4117..4133)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4126..4148)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 4275..4730
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4362..4381)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4572..4593
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 4756..4860
/gene="bla"
/label=AmpR promoter
CDS 4861..5721
/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(5079..5098)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5892..6480
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6381..6400
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 6634..6651
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 6768..6789
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 6804..6834
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6842..6858
/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 6847..6869
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 6866..6882
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 6866..6882
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 6901..6921
/label=T3
/note="T3 promoter, forward primer"
promoter 6903..6921
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 6949..7175
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 7176..7356
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 7403..7528
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 8021..8254
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 8439..8483
/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"
promoter 8772..8866
/label=H1-2O2 promoter
/note="doxycycline-inducible variant of the human H1 RNA
promoter (Henriksen et al., 2007)"
primer_bind 8783..8802
/label=H1
/note="Human H1 promoter, forward primer"
protein_bind 8820..8838
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 8848..8866
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
misc_feature 10784..10901
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 10950..501
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
ORIGIN
1 ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc tgggcgtggt
61 tccgggaaac gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc cgttcgcagc
121 gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc tgctccgccc
181 ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca
241 cgtctcacta gtaccctcgc agacggacag cgccagggag caatggcagc gcgccgaccg
301 cgatgggctg tggccaatag cggctgctca gcagggcgcg ccgagagcag cggccgggaa
361 ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg
421 tgttccgcat tctgcaagcc tccggagcgc acgtcggcag tcggctccct cgttgaccga
481 atcaccgacc tctctcccca gggggatctg tgagtttggg gacccttgat tgttctttct
541 ttttcgctat tgtaaaattc atgttatatg gagggggcaa agttttcagg gtgttgttta
601 gaatgggaag atgtcccttg tatcaccatg gaccctcatg ataattttgt ttctttcact
661 ttctactctg ttgacaacca ttgtctcctc ttattttctt ttcattttct gtaacttttt
721 cgttaaactt tagcttgcat ttgtaacgaa tttttaaatt cacttttgtt tatttgtcag
781 attgtaagta ctttctctaa tcactttttt ttcaaggcaa tcagggtata ttatattgta
841 cttcagcaca gttttagaga acaattgtta taattaaatg ataaggtaga atatttctgc
901 atataaattc tggctggcgt ggaaatattc ttattggtag aaacaactac atcctggtca
961 tcatcctgcc tttctcttta tggttacaat gatatacact gtttgagatg aggataaaat
1021 actctgagtc caaaccgggc ccctctgcta accatgttca tgccttcttc tttttcctac
1081 agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattgtaat
1141 acgactcact atagggcgaa ttgatatgtc tagattagat aaaagtaaag tgattaacag
1201 cgcattagag ctgcttaatg aggtcggaat cgaaggttta acaacccgta aactcgccca
1261 gaagctaggt gtagagcagc ctacattgta ttggcatgta aaaaataagc gggctttgct
1321 cgacgcctta gccattgaga tgttagatag gcaccatact cacttttgcc ctttagaagg
1381 ggaaagctgg caagattttt tacgtaataa cgctaaaagt tttagatgtg ctttactaag
1441 tcatcgcgat ggagcaaaag tacatttagg tacacggcct acagaaaaac agtatgaaac
1501 tctcgaaaat caattagcct ttttatgcca acaaggtttt tcactagaga atgcattata
1561 tgcactcagc gctgtggggc attttacttt aggttgcgta ttggaagatc aagagcatca
1621 agtcgctaaa gaagaaaggg aaacacctac tactgatagt atgccgccat tattacgaca
1681 agctatcgaa ttatttgatc accaaggtgc agagccagcc ttcttattcg gccttgaatt
1741 gatcatatgc ggattagaaa aacaacttaa atgtgaaagt gggtccgcgt acagcggctc
1801 ccgggagttc tagggatctg cccctctccc tccccccccc ctaacgttac tggccgaagc
1861 cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct
1921 tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt
1981 ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct
2041 ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc
2101 ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag
2161 gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc
2221 tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga
2281 tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt
2341 ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataaggatcc
2401 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
2461 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
2521 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
2581 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
2641 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
2701 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
2761 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
2821 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
2881 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
2941 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
3001 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
3061 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
3121 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
3181 gacgagttct tctgaggtac cttttttaag ggtgggcgcg ccgacccagc tttcttgtac
3241 aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc
3301 agtcaaaata aaatcattat ttgccatcca gctgatatcc gcccgcccca cgacccgcag
3361 cgcccgaccg aaaggagcgc acgaccccat gcatcggtac ctttaagacc aatgacttac
3421 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
3481 cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag
3541 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
3601 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
3661 tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt
3721 attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt
3781 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
3841 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
3901 tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
3961 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
4021 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta gggacgtacc
4081 caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg
4141 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
4201 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
4261 gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
4321 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
4381 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
4441 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
4501 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
4561 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
4621 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
4681 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
4741 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
4801 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
4861 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
4921 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
4981 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
5041 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
5101 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
5161 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
5221 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
5281 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
5341 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
5401 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
5461 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
5521 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
5581 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
5641 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
5701 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
5761 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
5821 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
5881 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
5941 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
6001 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
6061 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
6121 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
6181 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
6241 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
6301 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
6361 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
6421 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
6481 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
6541 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
6601 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
6661 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
6721 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
6781 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
6841 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc
6901 gcaattaacc ctcactaaag ggaacaaaag ctggagctgc aagcttaatg tagtcttatg
6961 caatactctt gtagtcttgc aacatggtaa cgatgagtta gcaacatgcc ttacaaggag
7021 agaaaaagca ccgtgcatgc cgattggtgg aagtaaggtg gtacgatcgt gccttattag
7081 gaaggcaaca gacgggtctg acatggattg gacgaaccac tgaattgccg cattgcagag
7141 atattgtatt taagtgccta gctcgataca taaacgggtc tctctggtta gaccagatct
7201 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
7261 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
7321 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
7381 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
7441 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
7501 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
7561 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
7621 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
7681 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
7741 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
7801 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
7861 gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg
7921 aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa
7981 agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt
8041 tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca
8101 gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc
8161 aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg
8221 ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac
8281 tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga
8341 tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa
8401 tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg
8461 aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata
8521 taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac
8581 tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc
8641 caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca
8701 gagacagatc cattcgatta gtgaacggat ctcgacggta tcgatcacga gactagcctc
8761 gagcggccgc aatatttgca tgtcgctatg tgttctggga aatcaccata aacgtgaaat
8821 ccctatcagt gatagagact tataagttcc ctatcagtga tagagacacc ggtgtgttgt
8881 aaatgagcac acaaaataca catgctaaaa tattatattc tatgaccttt ataaaatcaa
8941 ccaaaatctt ctttttaata actttagtat caataattag aatttttatg ttcctttttg
9001 caaactttta ataaaaatga gcaaaataaa aaaacgctag ttttagtaac tcgcgttgtt
9061 ttcttcacct ttaataatag ctactccacc acttgttcct aagcggtcag ctcctgcttc
9121 aatcattttt tgagcatctt caaatgttct aactccacca gctgctttaa ctaaagcatt
9181 gtctttaaca actgacttca ttagtttaac atcttcaaat gttgcacctg attttgaaaa
9241 tcctgttgat gttttaacaa attctaatcc agcttcaaca gctatttcac aagctttcat
9301 gatttcttct tttgttaata aacaattttc cataatacat ttaacaacat gtgatccagc
9361 tgcttttttt acagctttca tgtcttctaa aactaattca taatttttgt cttttaatgc
9421 accaatattt aataccatat caatttctgt tgcaccatct ttaattgctt cagaaacttc
9481 gaatgctttt gtagctgttg tgcatgcacc tagaggaaaa cctacaacat ttgttattcc
9541 tacatttgtg ccttttaata attctttaca atagcttgtt caatatgaat taacacaaac
9601 tgttgcaaaa tcaaattcaa ttgcttcatc acataattgt ttaatttcag ctttcgtagc
9661 atcttgtttt aataatgtgt gatctatata tttgtttagt ttcatttttt ctcctatata
9721 ttcattttta attttaattc tttaataatt tcgtctactt taactttagc gttttgaaca
9781 gattcaccaa cacctataaa ataaattttt agtttaggtt cagttccact tgggcgaaca
9841 gcaaatcatg acttatcttc taaataaaat tttagtaagt cttgtcctgg catattatac
9901 attccatcga tgtagtcttc aacattaaca actttaagtc cagcaatttg agttaagggt
9961 gttgctctca atgatttcat taatggttca atttttaatt tcttttcttc tggtttaaaa
10021 ttcaagttta aagtgaaagt gtaatatgca cccatttctt taaataaatc ttctaaatag
10081 tctactaatg ttttattttg ttttttataa aatcaagcag cctctgctat taatatagaa
10141 gcttgtattc catctttatc tctagctgag tcatcaatta catatccata actttcttca
10201 taagcaaaaa caaaatttaa tccgttatct tcttctttag caatttctct acccattcat
10261 ttaaatccag ttaaagtttt tacaatatta actccatatt tttcatgagc gattctatca
10321 cccaaatcac ttgttacaaa acttgaatat agagccggat tttttggaat gctatttaag
10381 cgttttagat ttgataattt tcaatcaatt aaaattggtc ctgtttgatt tccatctaat
10441 cttacaaaat gaccatcatg ttttattgcc attccaaatc tgtcagcatc tgggtcattc
10501 ataataataa tatctgcatc atgtttaata ccatattcaa gcggtatttt tcatgcagga
10561 tcaaattctg gatttggatt tacaacattt ttaaatgttt catcttcaaa tgcatgctct
10621 tcaacctcaa taacgttata tcctgattca cgtaatattt ttggggtaaa tttagttcct
10681 gttccattaa ctgcgctaaa aataattttt aaatcttttt tagcttcttg ctcttttttg
10741 tagaattctc gacctcgaga caaatggcag tattcatcca caattttaaa agaaaagggg
10801 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
10861 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
10921 gcagagatcc actttggccg cggctcgagg gggttgggg
//