| 质粒名称: | pCW57.1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 41393 |
| 存储实验室: | David Root |
|
载体骨架基本信息 |
|
| 载体名称: | pCW57.1 |
| 载体抗性: | 氯霉素和氨苄西林 |
| 空载体大小: | 9354 bp |
| 启动子： | TRE promoter, Tet ON |
| 载体类型: | 哺乳动物表达，慢病毒；网关目的载体，多西环素诱导 |
| 筛选标记: | 嘌呤霉素 |
| 宿生菌： | ccdB Survival |
| 生长条件: | 37 ℃ |
| 生长说明： | 注意这个质粒生长缓慢。液体培养可能需要培养两天才能大量生长。 |
|
插入基因信息 |
|
| 克隆方法: | 网关克隆 |
| 5' 测序引物 | LNCX |
| 3' 测序引物 |
O.PGK1b-R (GAACGGACGTGAAGAATGTG)
|
|
文献引用方法 |
|
| 材料和方法部分: | pCW57.1 was a gift from David Root (Addgene plasmid # 41393 ; http://n2t.net/addgene:41393 ; RRID:Addgene_41393) |
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 pCW57.1 9354 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION 3rd generation, Inducible lentiviral expression, TRE-gateway;
PGK-rtTA-2A-puro.
ACCESSION .
VERSION .
KEYWORDS pCW57.1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9354)
TITLE Root Lab pLEX and pLIX Gateway backbones
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 9354)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9354
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 11..29
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 47..65
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 83..101
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 118..136
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 154..172
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind 219..243
/label=LNCX
/note="Human CMV promoter, forward primer"
protein_bind 267..391
/gene="mutant version of attR"
/label=attR1
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
promoter 416..446
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
CDS 500..1159
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(566..585)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
CDS 1501..1806
/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 1685..1704
/label=ccdB-fwd
/note="ccdB gene, forward primer"
protein_bind complement(1847..1971)
/gene="mutant version of attR"
/label=attR2
/bound_moiety="LR Clonase(TM)"
/note="recombination site for the Gateway(R) LR reaction"
CDS 1989..2006
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
promoter 2016..2526
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
primer_bind 2445..2463
/label=hPGK-F
/note="Human PGK promoter, forward primer"
CDS 2536..3132
/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(2536..2555)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 3032..3052
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
CDS 3133..3186
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS 3187..3933
/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"
primer_bind complement(3973..3993)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4391..4402)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 4607..4840
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 4912..5033
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4949..4968)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5003..5022
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 5073..5208
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5135..5154
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 5208..5279
/gene="bla"
/label=AmpR promoter
CDS 5280..6140
/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(5498..5517)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6311..6899
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6800..6819
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7053..7070
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 7187..7208
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 7223..7253
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7261..7277
/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 7266..7288
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 7285..7301
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 7285..7301
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 7320..7340
/label=T3
/note="T3 promoter, forward primer"
promoter 7322..7340
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 7368..7594
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 7595..7775
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 7822..7947
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 8440..8673
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 9115..9232
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 9286..246
/label=tight TRE promoter
/note="Tet-responsive promoter PTight, consisting of seven
tet operator sequences followed by the minimal CMV
promoter"
protein_bind 9294..9312
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 9330..9348
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
1 tcgagtttac tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat
61 agagaacgta tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttatcc
121 ctatcagtga tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgtatgt
181 cgaggtaggc gtgtacggtg ggaggcctat ataagcagag ctcgtttagt gaaccgtcag
241 atcgcctgga gaattggcta gcatcaacaa gtttgtacaa aaaagctgaa cgagaaacgt
301 aaaatgatat aaatatcaat atattaaatt agattttgca taaaaaacag actacataat
361 actgtaaaac acaacatatc cagtcactat ggcggccgca ttaggcaccc caggctttac
421 actttatgct tccggctcgt ataatgtgtg gattttgagt taggatccgt cgagattttc
481 aggagctaag gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc
541 ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa
601 ccagaccgtt cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa
661 gttttatccg gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaattccg
721 tatggcaatg aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt
781 tttccatgag caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg
841 gcagtttcta cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt
901 ccctaaaggg tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac
961 cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg
1021 caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc
1081 cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga
1141 gtggcagggc ggggcgtaaa gatctggatc cggcttacta aaagccagat aacagtatgc
1201 gtatttgcgc gctgattttt gcggtataag aatatatact gatatgtata cccgaagtat
1261 gtcaaaaaga ggtatgctat gaagcagcgt attacagtga cagttgacag cgacagctat
1321 cagttgctca aggcatatat gatgtcaata tctccggtct ggtaagcaca accatgcaga
1381 atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga aaatcaggaa gggatggctg
1441 aggtcgcccg gtttattgaa atgaacggct cttttgctga cgagaacagg ggctggtgaa
1501 atgcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta
1561 cagagtgata ttattgacac gcccgggcga cggatggtga tccccctggc cagtgcacgt
1621 ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc
1681 tggcgcatga tgaccaccga tatggccagt gtgccggtct ccgttatcgg ggaagaagtg
1741 gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga
1801 atataaatgt caggctccct tatacacagc cagtctgcag gtcgaccata gtgactggat
1861 atgttgtgtt ttacagtatt atgtagtctg ttttttatgc aaaatctaat ttaatatatt
1921 gatatttata tcattttacg tttctcgttc agctttcttg tacaaagtgg tttagtaatg
1981 aaccggtcca ccaccaccac caccactaag gatccggggt tggggttgcg ccttttccaa
2041 ggcagccctg ggtttgcgca gggacgcggc tgctctgggc gtggttccgg gaaacgcagc
2101 ggcgccgacc ctgggtctcg cacattcttc acgtccgttc gcagcgtcac ccggatcttc
2161 gccgctaccc ttgtgggccc cccggcgacg cttcctgctc cgcccctaag tcgggaaggt
2221 tccttgcggt tcgcggcgtg ccggacgtga caaacggaag ccgcacgtct cactagtacc
2281 ctcgcagacg gacagcgcca gggagcaatg gcagcgcgcc gaccgcgatg ggctgtggcc
2341 aatagcggct gctcagcagg gcgcgccgag agcagcggcc gggaaggggc ggtgcgggag
2401 gcggggtgtg gggcggtagt gtgggccctg ttcctgcccg cgcggtgttc cgcattctgc
2461 aagcctccgg agcgcacgtc ggcagtcggc tccctcgttg accgaatcac cgacctctct
2521 ccccagcaat tcaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
2581 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
2641 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
2701 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
2761 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
2821 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
2881 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
2941 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
3001 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
3061 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
3121 aagcccggtg ccgaaggtag aggttctctc ctcacttgtg gtgatgttga agaaaaccct
3181 ggtccaatgt ctagactgga caagagcaaa gtcataaacg gagctctgga attactcaat
3241 ggtgtcggta tcgaaggcct gacgacaagg aaactcgctc aaaagctggg agttgagcag
3301 cctaccctgt actggcacgt gaagaacaag cgggccctgc tcgatgccct gccaatcgag
3361 atgctggaca ggcatcatac ccacttctgc cccctggaag gcgagtcatg gcaagacttt
3421 ctgcggaaca acgccaagtc ataccgctgt gctctcctct cacatcgcga cggggctaaa
3481 gtgcatctcg gcacccgccc aacagagaaa cagtacgaaa ccctggaaaa tcagctcgcg
3541 ttcctgtgtc agcaaggctt ctccctggag aacgcactgt acgctctgtc cgccgtgggc
3601 cactttacac tgggctgcgt attggaggaa caggagcatc aagtagcaaa agaggaaaga
3661 gagacaccta ccaccgattc tatgccccca cttctgagac aagcaattga gctgttcgac
3721 cggcagggag ccgaacctgc cttccttttc ggcctggaac taatcatatg tggcctggag
3781 aaacagctaa agtgcgaaag cggcgggccg accgacgccc ttgacgattt tgacttagac
3841 atgctcccag ccgatgccct tgacgacttt gaccttgata tgctgcctgc tgacgctctt
3901 gacgattttg accttgacat gctccccggg taactaagta aggatcgatc caagatatcg
3961 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
4021 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
4081 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
4141 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
4201 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
4261 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
4321 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
4381 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
4441 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
4501 ctccccgcct gtttcgcctc ggcgtccgga ctagaggtac ctttaagacc aatgacttac
4561 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
4621 cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag
4681 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
4741 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
4801 tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt
4861 attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt
4921 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
4981 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
5041 tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
5101 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
5161 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggcctt tcaaatatgt
5221 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
5281 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
5341 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
5401 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
5461 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
5521 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
5581 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
5641 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
5701 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
5761 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
5821 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
5881 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
5941 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
6001 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
6061 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
6121 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
6181 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
6241 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
6301 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
6361 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
6421 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
6481 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
6541 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
6601 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
6661 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
6721 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
6781 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
6841 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
6901 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
6961 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
7021 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
7081 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
7141 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
7201 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
7261 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
7321 caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc
7381 aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga
7441 gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg
7501 aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga
7561 tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg
7621 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
7681 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
7741 cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa
7801 gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg
7861 gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag
7921 aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg
7981 gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg
8041 gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc
8101 tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga
8161 tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac
8221 accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag
8281 caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
8341 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
8401 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
8461 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
8521 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
8581 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
8641 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
8701 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
8761 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
8821 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
8881 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
8941 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
9001 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
9061 aaccccgagg ggacaattct cgacctcgag acaaatggca gtattcatcc acaattttaa
9121 aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac
9181 agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta
9241 ttacagggac agcagagatc cactttggcc gcgaatcgat atgtcgagtt tactccctat
9301 cagtgataga gaacgtatgt cgagtttact ccctatcagt gatagagaac gatg
//
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