| 质粒名称: | pXD023 - pLKO-EF1a-Puro-P2A-FLuc-WPRE |
|---|---|
| 目录编号: | 192203 |
| 存储实验室: | Sidi Chen |
|
载体骨架基本信息 |
|
| 载体名称: | pLenti |
| 空载体大小: | 4038 bp |
| 完整质粒大小: | 9668 bp |
| 载体类型: | Lentiviral ; Mammalian Expression |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Lenti-PL(Puro-Luciferase) |
| 物种来源: | Synthetic |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | 未知 |
|
文献引用方法 |
|
| 材料和方法部分: | pXD023 - pLKO-EF1a-Puro-P2A-FLuc-WPRE was a gift from Sidi Chen (Addgene plasmid # 192203 ; http://n2t.net/addgene:192203 ; RRID:Addgene_192203) |
| 参考文献部分: | A genome-scale gain-of-function CRISPR screen in CD8 T cells identifies proline metabolism as a means to enhance CAR-T therapy. Ye L, Park JJ, Peng L, Yang Q, Chow RD, Dong MB, Lam SZ, Guo J, Tang E, Zhang Y, Wang G, Dai X, Du Y, Kim HR, Cao H, Errami Y, Clark P, Bersenev A, Montgomery RR, Chen S. Cell Metab. 2022 Apr 5;34(4):595-614.e14. doi: 10.1016/j.cmet.2022.02.009. Epub 2022 Mar 10. 10.1016/j.cmet.2022.02.009 PubMed 35276062 |
买家导航
pXD023 - pLKO-EF1a-Puro-P2A-FLuc-WPRE载体质粒基本信息
pXD023 - pLKO-EF1a-Puro-P2A-FLuc-WPRE质粒图谱载体图谱和pXD023 - pLKO-EF1a-Puro-P2A-FLuc-WPRE载体序列质粒序列多克隆位点信息
pXD023 - pLKO-EF1a-Puro-P2A-FLuc-WPRE质粒载体简介
pXD023 - pLKO-EF1a-Puro-P2A-FLuc-WPRE质粒序列载体序列
LOCUS pXD023 - pLKO-EF1a-Puro-P2A-FLuc-WPRE 9668 bp ds-DNA circular SYN 21-OCT-2022
DEFINITION Lenti-PL(Puro-Luciferase).
ACCESSION .
VERSION .
KEYWORDS pXD023 - pLKO-EF1a-Puro-P2A-FLuc-WPRE
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9668)
AUTHORS Ye L, Park JJ, Peng L, Yang Q, Chow RD, Dong MB, Lam SZ, Guo J, Tang
E, Zhang Y, Wang G, Dai X, Du Y, Kim HR, Cao H, Errami Y, Clark P,
Bersenev A, Montgomery RR, Chen S
TITLE A genome-scale gain-of-function CRISPR screen in CD8 T cells
identifies proline metabolism as a means to enhance CAR-T therapy.
JOURNAL Cell Metab. 2022 Apr 5;34(4):595-614.e14. doi:
10.1016/j.cmet.2022.02.009. Epub 2022 Mar 10.
PUBMED 35276062
REFERENCE 2 (bases 1 to 9668)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Oct 21, 2022 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..9668
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 5..231
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 232..412
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 459..584
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1077..1310
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1495..1539
/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 1741..1858
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 1997..3175
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 2228..3166
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 3123..3143
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
CDS 3188..3784
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(3188..3207)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 3684..3704
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
CDS 3794..3850
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS 3851..5503
/codon_start=1
/gene="luc+"
/product="firefly luciferase"
/label=luciferase
/note="enhanced luc+ version of the luciferase gene"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
primer_bind complement(3917..3935)
/label=LucNrev
/note="Firefly luciferase, reverse primer"
misc_feature 5525..6113
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(5578..5598)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(5996..6007)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 6185..6418
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 6496..6617
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6533..6552)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 6587..6606
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 6657..6792
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 6719..6738
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(6812..6831)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(6813..6831)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(6841..6858)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(6841..6857)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6850..6872)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 6999..7454
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(7086..7105)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 7296..7317
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 7480..7584
/gene="bla"
/label=AmpR promoter
CDS 7585..8445
/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(7803..7822)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 8616..9204
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 9105..9124
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 9358..9375
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 9492..9513
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 9528..9558
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 9566..9582
/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 9571..9593
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 9590..9606
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 9590..9606
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 9625..9645
/label=T3
/note="T3 promoter, forward primer"
promoter 9627..9645
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
1 ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat gagttagcaa
61 catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt aaggtggtac
121 gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg aaccactgaa
181 ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa cgggtctctc
241 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
301 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
361 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc
421 cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc
481 ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt
541 tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga
601 gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat
661 taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt
721 tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag
781 gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa
841 ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa
901 gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg
961 gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta
1021 gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga
1081 gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg
1141 ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg
1201 agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc
1261 caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg
1321 ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat
1381 aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac
1441 aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat
1501 gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca
1561 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
1621 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
1681 tttcagaccc acctcccaac cccgagggga cccgagacaa atggcagtat tcatccacaa
1741 ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat
1801 agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg
1861 ggtttattac agggacagca gagatccact ttggcgccgg ctcgaggggg cccgggtgca
1921 aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct tctaggtctt
1981 gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt
2041 ccccgagaag ttggggggag gggtcggcaa ttgatccggt gcctagagaa ggtggcgcgg
2101 ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga
2161 accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag
2221 aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg gttatggccc
2281 ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc ccgagcttcg
2341 ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc
2401 ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg
2461 cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc
2521 gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc acactggtat
2581 ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc
2641 gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg
2701 gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct
2761 ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctgcagg
2821 gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag
2881 gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc
2941 gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag gttgggggga
3001 ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc
3061 ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat
3121 tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgacgtac
3181 ggccaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag
3241 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
3301 tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
3361 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gccgtggcgg tctggaccac
3421 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
3481 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
3541 caaggagccc gcgtggttcc tggccaccgt cggagtctcg cccgaccacc agggcaaggg
3601 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
3661 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
3721 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
3781 tgccggatcc ggcgcaacaa acttctctct gctgaaacaa gccggagatg tcgaagagaa
3841 tcctggaccg atggaagacg ccaaaaacat aaagaaaggc ccggcgccat tctatccgct
3901 ggaagatgga accgctggag agcaactgca taaggctatg aagagatacg ccctggttcc
3961 tggaacaatt gcttttacag atgcacatat cgaggtggac atcacttacg ctgagtactt
4021 cgaaatgtcc gttcggttgg cagaagctat gaaacgatat gggctgaata caaatcacag
4081 aatcgtcgta tgcagtgaaa actctcttca attctttatg ccggtgttgg gcgcgttatt
4141 tatcggagtt gcagttgcgc ccgcgaacga catttataat gaacgtgaat tgctcaacag
4201 tatgggcatt tcgcagccta ccgtggtgtt cgtttccaaa aaggggttgc aaaaaatttt
4261 gaacgtgcaa aaaaagctcc caatcatcca aaaaattatt atcatggatt ctaaaacgga
4321 ttaccaggga tttcagtcga tgtacacgtt cgtcacatct catctacctc ccggttttaa
4381 tgaatacgat tttgtgccag agtccttcga tagggacaag acaattgcac tgatcatgaa
4441 ctcctctgga tctactggtc tgcctaaagg tgtcgctctg cctcatagaa ctgcctgcgt
4501 gagattctcg catgccagag atcctatttt tggcaatcaa atcattccgg atactgcgat
4561 tttaagtgtt gttccattcc atcacggttt tggaatgttt actacactcg gatatttgat
4621 atgtggattt cgagtcgtct taatgtatag atttgaagag gagctgtttc tgaggagcct
4681 tcaggattac aagattcaaa gtgcgctgct ggtgccaacc ctattctcct tcttcgccaa
4741 aagcactctg attgacaaat acgatttatc taatttacac gaaattgctt ctggtggcgc
4801 tcccctctct aaggaagtcg gggaagcggt tgccaagagg ttccatctgc caggtatcag
4861 gcaaggatat gggctcactg agactacatc agctattctg attacacccg agggggatga
4921 taaaccgggc gcggtcggta aagttgttcc attttttgaa gcgaaggttg tggatctgga
4981 taccgggaaa acgctgggcg ttaatcaaag aggcgaactg tgtgtgagag gtcctatgat
5041 tatgtccggt tatgtaaaca atccggaagc gaccaacgcc ttgattgaca aggatggatg
5101 gctacattct ggagacatag cttactggga cgaagacgaa cacttcttca tcgttgaccg
5161 cctgaagtct ctgattaagt acaaaggcta tcaggtggct cccgctgaat tggaatccat
5221 cttgctccaa caccccaaca tcttcgacgc aggtgtcgca ggtcttcccg acgatgacgc
5281 cggtgaactt cccgccgccg ttgttgtttt ggagcacgga aagacgatga cggaaaaaga
5341 gatcgtggat tacgtcgcca gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt
5401 gtttgtggac gaagtaccga aaggtcttac cggaaaactc gacgcaagaa aaatcagaga
5461 gatcctcata aaggccaaga agggcggaaa gatcgccgtg taagaattca cgcgttaagt
5521 cgacaatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
5581 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
5641 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
5701 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
5761 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
5821 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
5881 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
5941 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
6001 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
6061 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcgtcgact
6121 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg
6181 ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg
6241 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
6301 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
6361 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
6421 acgtatagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata
6481 tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc
6541 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
6601 ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccatcc
6661 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
6721 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
6781 tttttggagg cctagggacg tacccaattc gccctatagt gagtcgtatt acgcgcgctc
6841 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
6901 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
6961 cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt
7021 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
7081 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
7141 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
7201 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
7261 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
7321 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
7381 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
7441 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
7501 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
7561 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
7621 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
7681 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
7741 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
7801 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
7861 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
7921 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
7981 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
8041 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
8101 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
8161 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
8221 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
8281 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
8341 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
8401 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
8461 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
8521 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
8581 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
8641 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
8701 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
8761 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
8821 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
8881 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
8941 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
9001 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
9061 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
9121 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
9181 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
9241 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
9301 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
9361 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
9421 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
9481 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
9541 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
9601 tatgaccatg attacgccaa gcgcgcaatt aaccctcact aaagggaaca aaagctggag
9661 ctgcaagc
//
