| 质粒名称: | pLV-EF1a-FRB-mCherry-MinD-IRES-Puro |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 213116 |
| 存储实验室: | Scott Coyle |
|
载体骨架基本信息 |
|
| 载体名称: | pLV-EF1a |
| 载体出品公司: | Clontech |
| 空载体大小: | 8699 bp |
| 完整质粒大小: | 10526 bp |
| 载体类型: | 哺乳细胞表达, Lentiviral |
| 筛选标记: | Puromycin |
| 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: | FRB-mCherry-MinD |
| 物种来源: | Escherichia coli |
| 插入大小: | 1827 bp |
| 启动子: | EF1a |
| 克隆方法: |
Gibson Cloning
|
| 5’测序引物 | EF1a_F (TCAAGCCTCAGACAGTGGTTC) |
| 3’测序引物 | EF1a_R (TAGAGGGAAACCGTTGCTAGC) |
|
文献引用方法 |
|
| 材料和方法部分: |
pLV-EF1a-FRB-mCherry-MinD-IRES-Puro was a gift from Scott Coyle (Addgene plasmid # 213116 ; http://n2t.net/addgene:213116 ; RRID:Addgene_213116)
|
| 参考文献部分: | A programmable reaction-diffusion system for spatiotemporal cell signaling circuit design. Rajasekaran R, Chang CC, Weix EWZ, Galateo TM, Coyle SM. Cell. 2023 Dec 27:S0092-8674(23)01339-9. doi: 10.1016/j.cell.2023.12.007. 10.1016/j.cell.2023.12.007 PubMed 38181787 |
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 pLV-EF1a-FRB-mCherry-MinD-IRES-Puro 10526 bp ds-DNA circular SYN 26-JAN-2024
DEFINITION Expression of FRB-mCherry-MinD in mammalian cells for payload
delivery.
ACCESSION .
VERSION .
KEYWORDS pLV-EF1a-FRB-mCherry-MinD-IRES-Puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10526)
AUTHORS Rajasekaran R, Chang CC, Weix EWZ, Galateo TM, Coyle SM
TITLE A programmable reaction-diffusion system for spatiotemporal cell
signaling circuit design.
JOURNAL Cell. 2023 Dec 27:S0092-8674(23)01339-9. doi:
10.1016/j.cell.2023.12.007.
PUBMED 38181787
REFERENCE 2 (bases 1 to 10526)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jan 26, 2024 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..10526
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(286..330)
/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 515..748
/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 1245..1370
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
LTR 1417..2050
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
polyA_signal 2218..2352
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2242..2261)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2296..2315
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
promoter 2400..2504
/gene="bla"
/label=AmpR promoter
CDS 2505..3365
/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(2723..2742)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 3536..4124
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4025..4044
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4278..4295
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 4412..4433
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4448..4478
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4486..4502
/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 4491..4513
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 4510..4526
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 4510..4526
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
LTR 4654..5287
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 5494..6082
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS 5600..5611
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind 6009..6029
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(6106..6705)
/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(6189..6209)
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
primer_bind 6686..6705
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind complement(6867..6886)
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
primer_bind 7096..7113
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
CDS complement(8204..8911)
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(8479..8498)
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind 8748..8766
/label=mCherry-R
/note="mCherry, reverse primer"
CDS complement(8912..9181)
/codon_start=1
/product="FKBP-rapamycin binding domain of human FRAP"
/label=FRB
/translation="EMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETS
FNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISKQ"
regulatory 9181..9190
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
intron 9206..10144
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind complement(9229..9249)
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
promoter complement(10145..10356)
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
misc_feature 10434..23
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1 (lacking the first T)"
ORIGIN
1 ccaatccccc cttttctttt aaaggcgata ccgtcgagat ccgttcacta atcgaatgga
61 tctgtctctg tctctctctc caccttcttc ttctattcct tcgggcctgt cgggtcccct
121 cggggttggg aggtgggtct gaaacgataa tggtgaatat ccctgcctaa ctctattcac
181 tatagaaagt acagcaaaaa ctattcttaa acctaccaag cctcctacta tcattatgaa
241 taattttata taccacagcc aatttgttat gttaaaccaa ttccacaaac ttgcccattt
301 atctaattcc aataattctt gttcattctt ttcttgctgg ttttgcgatt cttcaattaa
361 ggagtgtatt aagcttgtgt aattgttaat ttctctgtcc cactccatcc aggtcgtgtg
421 attccaaatc tgttccagag atttattact ccaactagca ttccaaggca cagcagtggt
481 gcaaatgagt tttccagagc aaccccaaat ccccaggagc tgttgatcct ttaggtatct
541 ttccacagcc aggattcttg cctggagctg cttgatgccc cagactgtga gttgcaacag
601 atgctgttgc gcctcaatag ccctcagcaa attgttctgc tgctgcacta taccagacaa
661 taattgtctg gcctgtaccg tcagcgtcat tgacgctgcg cccatagtgc ttcctgctgc
721 tcccaagaac ccaaggaaca aagctcctat tcccactgct cttttttctc tctgcaccac
781 tcttctcttt gccttggtgg gtgctactcc taatggttca atttttacta ctttatattt
841 atataattca cttctccaat tgtccctcat atctcctcct ccaggtctga agatcagcgg
901 ccggccgctt gctgtgcggt ggtcttactt ttgttttgct cttcctctat cttgtctaaa
961 gcttccttgg tgtcttttat ctctatcctt tgatgcacac aatagagggt tgctactgta
1021 ttatataatg atctaagttc ttctgatcct gtctgaaggg atggttgtag ctgtcccagt
1081 atttgtctac agccttctga tgtttctaac aggccaggat taactgcgaa tcgttctagc
1141 tccctgcttg cccatactat atgttttaat ttatattttt tctttccccc tggccttaac
1201 cgaatttttt cccatcgcga tctaattctc ccccgcttaa tactgacgct ctcgcaccca
1261 tctctctcct tctagcctcc gctagtcaaa atttttggcg tactcaccag tcgccgcccc
1321 tcgcctcttg ccgtgcgcgc ttcagcaagc cgagtcctgc gtcgagagag ctcctctggt
1381 ttccctttcg ctttcaagtc cctgttcggg cgccactgct agagattttc cacactgact
1441 aaaagggtct gagggatctc tagttaccag agtcacacaa cagacgggca cacactactt
1501 gaagcactca aggcaagctt tattgaggct taagcagtgg gttccctagt tagccagaga
1561 gctcccaggc tcagatctgg tctaaccaga gagacccagt acaggcaaaa agcagctgct
1621 tatatgcagg atctgagggc tcgccactcc ccagtcccgc ccaggccacg cctccctgga
1681 aagtccccag cggaaagtcc cttgtagcaa gctcgatatc agcagttctt gaagtactcc
1741 ggatgcagct ctcgggccac gtgatgaaat gctaggcggc tgtcaaacct ccactctaac
1801 acttctctct ccgggtcatc catcccatgc aggctcacag ggtgtaacaa gctggtgttc
1861 tctcctttat tggcctcttc taccttatct ggctcaactg gtactagctt gtagcaccat
1921 ccaaaggtca gtggatatct gacccctggc cctggtgtgt agttctgcta atcagggaag
1981 tagccttgtg tgtggtagat ccacagatca aggatatctt gtcttctttg ggagtgaatt
2041 agcccttcca actactaagt ttgtagtaca tatttaacaa atacaatttc tttaaaatga
2101 aaataattca gaggaatcac aggtttagag taaatgaaac cacaggtaat tggcagtggt
2161 aatagggtat ggggtgggaa gtttgggatg attttggtta gcttgagtta tccagttgat
2221 ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
2281 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
2341 aataaacaag ttgatctccc gatccgtcga cgtcaggtgg cacttttcgg ggaaatgtgc
2401 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
2461 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
2521 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
2581 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
2641 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
2701 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
2761 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
2821 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
2881 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
2941 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
3001 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
3061 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
3121 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
3181 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
3241 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
3301 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
3361 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
3421 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
3481 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
3541 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
3601 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
3661 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
3721 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
3781 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
3841 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
3901 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
3961 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
4021 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
4081 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
4141 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
4201 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
4261 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
4321 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
4381 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
4441 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
4501 aatttcacac aggaaacagc tatgaccatg attacgccaa gctctagcta gaggtcgacg
4561 gtaaacgcta gcaatgtcaa ggcctctcac tctctgatat tcatttcttt gcaagttata
4621 aatactgaat aataagatga catgaactac tactgctaga gattttccac actgactaaa
4681 agggtctgag ggatctctag ttaccagagt cacacaacag acgggcacac actacttgaa
4741 gcactcaagg caagctttat tgaggcttaa gcagtgggtt ccctagttag ccagagagct
4801 cccaggctca gatctggtct aaccagagag acccagtaca ggcaaaaagc agctgcttat
4861 atgcaggatc tgagggctcg ccactcccca gtcccgccca ggccacgcct ccctggaaag
4921 tccccagcgg aaagtccctt gtagcaagct cgatatcagc agttcttgaa gtactccgga
4981 tgcagctctc gggccacgtg atgaaatgct aggcggctgt caaacctcca ctctaacact
5041 tctctctccg ggtcatccat cccatgcagg ctcacagggt gtaacaagct ggtgttctct
5101 cctttattgg cctcttctac cttatctggc tcaactggta ctagcttgta gcaccatcca
5161 aaggtcagtg gatatctgac ccctggccct ggtgtgtagt tctgctaatc agggaagtag
5221 ccttgtgtgt ggtagatcca cagatcaagg atatcttgtc ttcgttggga gtgaattagc
5281 ccttccagtc ccctcttttc ttttaaaaag tggctaagat ctacagctgc cttgtaagtc
5341 attggtctta aaggtacctg aggtgtgact ggaaaaccca cctcctcctc ctcttgtgct
5401 tctagccagg cacaatcagc attggtagct gctgtattgc tacttgtgat tgctccatgt
5461 ttttctaggt ctcgactgca gaattaattc caggcgggga ggcggcccaa agggagatcc
5521 gactcgtctg agggcgaagg cgaagacgcg gaagaggccg cagagccggc agcaggccgc
5581 gggaaggaag gtccgctgga ttgagggccg aagggacgta gcagaaggac gtcccgcgca
5641 gaatccaggt ggcaacacag gcgagcagcc atggaaagga cgtcagcttc cccgacaaca
5701 ccacggaatt gtcagtgccc aacagccgag cccctgtcca gcagcgggca aggcaggcgg
5761 cgatgagttc cgccgtggca atagggaggg ggaaagcgaa agtcccggaa aggagctgac
5821 aggtggtggc aatgccccaa ccagtggggg ttgcgtcagc aaacacagtg cacaccacgc
5881 cacgttgcct gacaacgggc cacaactcct cataaagaga cagcaaccag gatttataca
5941 aggaggagaa aatgaaagcc atacgggaag caatagcatg atacaaaggc attaaagcag
6001 cgtatccaca tagcgtaaaa ggagcaacat agttaagaat accagtcaat ctttcacaaa
6061 ttttgtaatc cagaggttga ttgttccaga cgcgccccgc ccgggtcagg caccgggctt
6121 gcgggtcatg caccaggtgc gcggtccttc gggcacctcg acgtcggcgg tgacggtgaa
6181 gccgagccgc tcgtagaagg ggaggttgcg gggcgcggag gtctccagga aggcgggcac
6241 cccggcgcgc tcggccgcct ccactccggg gagcacgacg gcgctgccca gacccttgcc
6301 ctggtggtcg ggcgagacgc cgacggtggc caggaaccac gcgggctcct tgggccggtg
6361 cggcgccagg aggccttcca tctgttgctg cgcggccagc cgggaaccgc tcaactcggc
6421 catgcgcggg ccgatctcgg cgaacaccgc ccccgcttcg acgctctccg gcgtggtcca
6481 gaccgccacc gcggcgccgt cgtccgcgac ccacaccttg ccgatgtcga gcccgacgcg
6541 cgtgaggaag agttcttgca gctcggtgac ccgctcgatg tggcggtccg gatcgacggt
6601 gtggcgcgtg gcggggtagt cggcgaacgc ggcggcgagg gtgcgtacgg ccctggggac
6661 gtcgtcgcgg gtggcgaggc gcaccgtggg cttgtactcg gtcatggtat tatcgtgttt
6721 ttcaaaggaa aaccacgtcc ccgtggttcg gggggcctag acgttttttt aacctcgact
6781 aaacacatgt aaagcatgtg caccgaggcc ccagatcaga tcccatacaa tggggtacct
6841 tctgggcatc cttcagcccc ttgttgaata cgcttgagga gagccatttg actctttcca
6901 caactatcca actcacaacg tggcactggg gttgtgccgc ctttgcaggt gtatcttata
6961 cacgtggctt ttggccgcag aggcacctgt cgccaggtgg ggggttccgc tgcctgcaaa
7021 gggtcgctac agacgttgtt tgtcttcaag aagcttccag aggaactgct tccttcacga
7081 cattcaacag accttgcatt cctttggcga gaggggaaag acccctagga atgctcgtca
7141 agaagacagg gccaggtttc cgggccctca cattgccaaa agacggcaat atggtggaaa
7201 ataacatata gacaaacgca caccggcctt attccaagcg gcttcggcca gtaacgttag
7261 gggggggggg cggaattgat cccgctagag ggaaaccgtt gctagcttaa gttagttaac
7321 aattgttacc gcggcccgtc gactctagag cggccgctta tcccccgaac agccgtttaa
7381 ggaatccctt cttttcctcc tctatgaagc ggaagggccg ttcttccccg aggagccgtt
7441 ctaccgtatc ggcataagct ttcccagcat ccgcatttat gtcaagtatg acaggctccc
7501 cctgattaga tgcgcggagg acggactggt cttccgggat aacgcctacc agttttatcc
7561 tcagaatctc gaggacatct tccattgaaa gcatatctcc ccgagatact cgaccggggt
7621 tatatcgggt aagcaggaga tgctccttaa ttggctcctc cccgttttct gccctccgtg
7681 atttgctggc gagtatcccc agtattcggt ctgaatctcg cacactagac acctccggat
7741 tagttgtgat tatggcttca tctgcgaagt acagtgccat aagtgcccct gtctctatac
7801 cggcgggaga gtcacaaact atgaactcaa agtccatcgc tttcaagtca tccaaaactt
7861 ttgcgacacc ctccctggtg agcgcgtctt tgtcccgcgt ttgtgaggca ggaagaatat
7921 acaagttttc ggttcgttta tcttttataa gtgcctgatt aagagtcgcg tcgccttgga
7981 tgacattgac aaaatcgtag acaacgcgcc gttcgcagcc cattatcagg tccaggttcc
8041 tgagcccaat atcaaaatct atgaccacag ttttcttgcc tttctgtgcg aggccggtag
8101 ctatggctgc gctggatgtg gtttttccaa ctccgccctt acctgaagtc acaactatta
8161 tgcgggccat ggtacccgca gagccagcac ttccggcgct gtccttgtac agctcgtcca
8221 tgccgccggt ggagtggcgg ccctcggcgc gttcgtactg ttccacgatg gtgtagtcct
8281 cgttgtggga ggtgatgtcc aacttgatgt tgacgttgta ggcgccgggc agctgcacgg
8341 gcttcttggc cttgtaggtg gtcttgacct cagcgtcgta gtggccgccg tccttcagct
8401 tcagcctctg cttgatctcg cccttcaggg cgccgtcctc ggggtacatc cgctcggagg
8461 aggcctccca gcccatggtc ttcttctgca ttacggggcc gtcggagggg aagttggtgc
8521 cgcgcagctt caccttgtag atgaactcgc cgtcctgcag ggaggagtcc tgggtcacgg
8581 tcaccacgcc gccgtcctcg aagttcatca cgcgctccca cttgaagccc tcggggaagg
8641 acagcttcaa gtagtcgggg atgtcggcgg ggtgcttcac gtaggccttg gagccgtaca
8701 tgaactgagg ggacaggatg tcccaggcga agggcagggg gccacccttg gtcaccttca
8761 gcttggcggt ctgggtgccc tcgtaggggc ggccctcgcc ctcgccctcg atctcgaact
8821 cgtggccgtt cacggagccc tccatgtgca ccttgaagcg catgaactcc ttgatgatgg
8881 ccatgttatc ctcctcgccc ttgctcacca tctgctttga gattcgtcgg aacacatgat
8941 aatagaggtc ccaggcttgg gtgaggtcct tgacattccc tgatttcatg tacttcctgc
9001 accactcttg ggcctccatt aaatctcgac cataggcctg attaaaggat gtttccttca
9061 gagtctgggg gccccgttcc atcatagcat gcaagggctc cagcacctca aacatgcctt
9121 tcacgttcct ttccccaaag tacaaacgag atgcctcttc caggccttca tgccacatct
9181 ccatggtggc ggatcctcac gacacctgaa atggaagaaa aaaactttga accactgtct
9241 gaggcttgag aatgaaccaa gatccaaact caaaaagggc aaattccaag gagaattaca
9301 tcaagtgcca agctggccta acttcagtct ccacccactc agtgtgggga aactccatcg
9361 cataaaaccc ctccccccaa cctaaagacg acgtactcca aaagctcgag aactaatcga
9421 ggtgcctgga cggcgcccgg tactccgtgg agtcacatga agcgacggct gaggacggaa
9481 aggccctttt cctttgtgtg ggtgactcac ccgcccgctc tcccgagcgc cgcgtcctcc
9541 attttgagct ccctgcagca gggccgggaa gcggccatct ttccgctcac gcaactggtg
9601 ccgaccgggc cagccttgcc gcccagggcg gggcgataca cggcggcgcg aggccaggca
9661 ccagagcagg ccggccagct tgagactacc cccgtccgat tctcggtggc cgcgctcgca
9721 ggccccgcct cgccgaacat gtgcgctggg acgcacgggc cccgtcgccg cccgcggccc
9781 caaaaaccga aataccagtg tgcagatctt ggcccgcatt tacaagacta tcttgccaga
9841 aaaaaagcgt cgcagcaggt catcaaaaat tttaaatggc tagagactta tcgaaagcag
9901 cgagacaggc gcgaaggtgc caccagattc gcacgcggcg gccccagcgc ccaggccagg
9961 cctcaactca agcacgaggc gaaggggctc cttaagcgca aggcctcgaa ctctcccacc
10021 cacttccaac ccgaagctcg ggatcaagaa tcacgtactg cagccaggtg gaagtaattc
10081 aaggcacgca agggccataa cccgtaaaga ggccaggccc gcgggaacca cacacggcac
10141 ttacctgtgt tctggcggca aacccgttgc gaaaaagaac gttcacggcg actactgcac
10201 ttatatacgg ttctccccca ccctcgggaa aaaggcggag ccagtacacg acatcacttt
10261 cccagtttac cccgcgccac cttctctagg caccggttca attgccgacc cctcccccca
10321 acttctcggg gactgtgggc gatgtgcgct ctgcccactg acgggcacct cgagtgaaga
10381 cgaaagggcc tcatcgataa actggatctc tgctgtccct gtaataaacc cgaaaatttt
10441 gaatttttgt aatttgtttt tgtaattctt tagtttgtat gtctgttgct attatgtcta
10501 ctattctttc ccctgcactg tacccc
//
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