LOCUS       Plasmid_249545_p        9477 bp DNA     circular SYN 04-FEB-2026
DEFINITION  Backbone for cloning LwCas13a guides under U6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9477)
  AUTHORS   Abudayyeh OO, Gootenberg JS, Essletzbichler P, Han S, Joung J, 
            Belanto JJ, Verdine V, Cox DBT, Kellner MJ, Regev A, Lander ES, 
            Voytas DF, Ting AY, Zhang F
  TITLE     RNA targeting with CRISPR-Cas13.
  JOURNAL   Nature. 2017 Oct 4. doi: 10.1038/nature24049.
  PUBMED    28976959
REFERENCE   2  (bases 1 to 9477)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 5, 2026 from SnapGene 7.2.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            2017 Oct 4. doi: 10.1038/nature24049."
COMMENT     Alias: 18-MS2-U6-GAPDH-ASAP2
FEATURES             Location/Qualifiers
     source          1..9477
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          6840..6841
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     86..103
                     /label=U6-F1
     rep_origin      138..726
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     627..646
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    789..801
                     /label=scaffold-0
     misc_RNA        806..824
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_feature    835..870
                     /label=scaffold-1
     primer_bind     complement(835..853)
                     /label=MS2-scaffold-R-2
     misc_RNA        876..894
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     primer_bind     complement(923..966)
                     /label=MS2-scaffold-R-2
     misc_feature    932..975
                     /label=scaffold-2
     promoter        985..1225
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     985..1005
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1156..1175
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     1222..1271
                     /label=left arm F
     primer_bind     complement(1222..1271)
                     /label=Plasmid left
     misc_feature    1246..1744
                     /label=5' arm
     primer_bind     1590..1609
                     /label=GAPDH-5 arm F
     primer_bind     complement(1720..1769)
                     /label=left arm R
     primer_bind     1720..1769
                     /label=recoded exon 9 F
     misc_feature    1745..7118
                     /label=Cargo
     misc_feature    1745..1808
                     /label=recoded Exon 9
     primer_bind     complement(1787..1808)
                     /label=Recoded exone9 R
     primer_bind     1789..1808
                     /label=Exon9 F
     CDS             1809..1862
                     /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"
     primer_bind     1838..1887
                     /label=Insert-F
     primer_bind     complement(1838..1887)
                     /label=Vector-R
     primer_bind     complement(1838..1862)
                     /label=recoded exon 9 R
     primer_bind     complement(1844..1862)
                     /label=载体.反向
     CDS             1863..1883
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1908..6008
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     primer_bind     complement(2288..2307)
                     /label=Exon9-R
     primer_bind     2785..2809
                     /label=Add-1
     primer_bind     3496..3520
                     /label=Add-2
     primer_bind     complement(3638..3657)
                     /label=Add-1 R
     primer_bind     4222..4246
                     /label=Add-3
     primer_bind     4308..4327
                     /label=Cas9-F
     primer_bind     complement(4345..4364)
                     /label=Add-2 R
     primer_bind     complement(4746..4765)
                     /label=Cas9-R
     primer_bind     4942..4966
                     /label=Add-4
     primer_bind     complement(5054..5073)
                     /label=Add-3 R
     primer_bind     complement(5511..5531)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     5650..5674
                     /label=Add-5
     primer_bind     complement(5733..5752)
                     /label=Add-4 R
     primer_bind     5984..6003
                     /label=引物 1
     CDS             6009..6056
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6072..6125
                     /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             join(6126..6128,6129..6839)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(6168..6189)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6424..6443)
                     /label=Add-5 R
     primer_bind     complement(6429..6448)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(6514..6530)
                     /label=Add-5 R
     primer_bind     6633..6652
                     /label=Gapdh-3 Arm F
     primer_bind     6776..6797
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(6815..6864)
                     /label=Insert-R
     primer_bind     6815..6864
                     /label=Vector-F
     primer_bind     complement(6820..6839)
                     /label=引物 2
     primer_bind     6840..6864
                     /label=poly signal F
     primer_bind     complement(6840..6864)
                     /label=ASAP2 R
     polyA_signal    6894..7118
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7094..7143)
                     /label=poly signal R
     primer_bind     7094..7143
                     /label=right arm F
     misc_feature    7119..7618
                     /label=3' arm
     primer_bind     complement(7360..7379)
                     /label=GAPDH-3 Arm R
     primer_bind     complement(7593..7642)
                     /label=right arm R
     primer_bind     7593..7642
                     
     primer_bind     complement(7658..7675)
                     /label=U6--R1
     primer_bind     7676..7693
                     /label=U6--F1
     rep_origin      7742..8197
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7829..7848)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8039..8060
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8214..8233)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8333..8355
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8393..8411)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     8433..8451
                     /label=U6-F
     promoter        8479..8583
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8584..8652,8653..9444)
                     /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(8802..8821)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
       61 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
      121 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
      181 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
      241 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
      301 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
      361 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
      421 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
      481 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
      541 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
      601 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
      661 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
      721 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
      781 cagctagcgt tttagagcta ggccaacatg aggatcaccc atgtctgcag ggcctagcaa
      841 gttaaaataa ggctagtccg ttatcaactt ggccaacatg aggatcaccc atgtctgcag
      901 ggccaagtgg caccgagtcg gtgctttttt tgttttagag ctagaaatag caagttaaaa
      961 taaggctagt ccgtttttac cggtgagggc ctatttccca tgattccttc atatttgcat
     1021 atacgataca aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata
     1081 ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa
     1141 ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg
     1201 gctttatata tcttgtggaa aggacgaaac accgatttag actacaagac tgtggatggc
     1261 ccctccggga aactgtggcg tgatggccgc ggggctctcc agaacatcat ccctgcctct
     1321 actggcgctg ccaaggctgt gggcaaggtc atccctgagc tgaacgggaa gctcactggc
     1381 atggccttcc gtgtccccac tgccaacgtg tcagtggtgg acctgacctg ccgtctagaa
     1441 aaacctgcca aatatgatga catcaagaag gtggtgaagc aggcgtcgga gggccccctc
     1501 aagggcatcc tgggctacac tgagcaccag gtggtctcct ctgacttcaa cagcgacacc
     1561 cactcctcca cctttgacgc tggggctggc attgccctca acgaccactt tgtcaagctc
     1621 atttcctggt atgtggctgg ggccagagac tggctcttaa aaagtgcagg gtctggcgcc
     1681 ctctggtggc tggctcagaa aaagggccct gacaactctt tacatcttct aggtatgaca
     1741 acgagttcgg atatagcaat agagtggtcg atctgatggc tcatatggct agcaaagagg
     1801 gaagcggaga aggtagaggt tctctcctca cttgtggtga tgttgaagaa aaccctggtc
     1861 caccaaagaa gaagcggaag gtcggtatcc acggagtccc agcagccgac aagaagtaca
     1921 gcatcggcct ggacatcggc accaactctg tgggctgggc cgtgatcacc gacgagtaca
     1981 aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc atcaagaaga
     2041 acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga
     2101 gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg caagagatct
     2161 tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa gagtccttcc
     2221 tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc gtggacgagg
     2281 tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg gtggacagca
     2341 ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc aagttccggg
     2401 gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac aagctgttca
     2461 tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg
     2521 tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg gaaaatctga
     2581 tcgcccagct gcccggcgag aagaagaatg gcctgttcgg aaacctgatt gccctgagcc
     2641 tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc aaactgcagc
     2701 tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc ggcgaccagt
     2761 acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg agcgacatcc
     2821 tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc aagagatacg
     2881 acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag ctgcctgaga
     2941 agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac attgacggcg
     3001 gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag atggacggca
     3061 ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag cggaccttcg
     3121 acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt ctgcggcggc
     3181 aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag atcctgacct
     3241 tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc gcctggatga
     3301 ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg gacaagggcg
     3361 cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg cccaacgaga
     3421 aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac gagctgacca
     3481 aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa
     3541 aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag cagctgaaag
     3601 aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc gtggaagatc
     3661 ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact
     3721 tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt
     3781 ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca
     3841 aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc
     3901 tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg
     3961 acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag
     4021 aggacatcca gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca
     4081 atctggccgg cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg
     4141 agctcgtgaa agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag
     4201 agaaccagac cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag
     4261 agggcatcaa agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc
     4321 tgcagaacga gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc
     4381 aggaactgga catcaaccgg ctgtccgact acgatgtgga ccatatcgtg cctcagagct
     4441 ttctgaagga cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca
     4501 agagcgacaa cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc
     4561 tgctgaacgc caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag
     4621 gcggcctgag cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc
     4681 agatcacaaa gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga
     4741 atgacaagct gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt
     4801 tccggaagga tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg
     4861 acgcctacct gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa
     4921 gcgagttcgt gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg
     4981 agcaggaaat cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt
     5041 tcaagaccga gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa
     5101 acggcgaaac cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag
     5161 tgctgagcat gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca
     5221 gcaaagagtc tatcctgccc aagaggaaca gcgataagct gatcgccaga aagaaggact
     5281 gggaccctaa gaagtacggc ggcttcgaca gccccaccgt ggcctattct gtgctggtgg
     5341 tggccaaagt ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga
     5401 tcaccatcat ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg
     5461 gctacaaaga agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc
     5521 tggaaaacgg ccggaagaga atgctggcct ctgccggcga actgcagaag ggaaacgaac
     5581 tggccctgcc ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga
     5641 agggctcccc cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc
     5701 tggacgagat catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta
     5761 atctggacaa agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg
     5821 ccgagaatat catccacctg tttaccctga ccaatctggg agcccctgcc gccttcaagt
     5881 actttgacac caccatcgac cggaagaggt acaccagcac caaagaggtg ctggacgcca
     5941 ccctgatcca ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg
     6001 gaggcgacaa aaggccggcg gccacgaaaa aggccggcca ggcaaaaaag aaaaaggaat
     6061 tcggcagtgg agagggcaga ggaagtctgc taacatgcgg tgacgtcgag gagaatcctg
     6121 gcccagtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     6181 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     6241 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     6301 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     6361 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     6421 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     6481 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     6541 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     6601 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     6661 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     6721 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     6781 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     6841 gagcggccgc gtcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     6901 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     6961 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     7021 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     7081 caatagcagg catgctgggg atgcggtggg ctctatggat ttggctacag caacagggtg
     7141 gtggacctca tggcccacat ggcctccaag gagtaagacc cctggaccac cagccccagc
     7201 aagagcacaa gaggaagaga gagaccctca ctgctgggga gtccctgcca cactcagtcc
     7261 cccaccacac tgaatctccc ctcctcacag ttgccatgta gaccccttga agaggggagg
     7321 ggcctaggga gccgcacctt gtcatgtacc atcaataaag taccctgtgc tcaaccagtt
     7381 acttgtcctg tcttattcta gggtctgggg cagaggggag ggaagctggg cttgtgtcaa
     7441 ggtgagacat tcttgctggg gagggacctg gtatgttctc ctcagactga gggtagggcc
     7501 tccaaacagc cttgcttgct tcgagaacca tttgcttccc gctcagacgt cttgagtgct
     7561 acaggaagct ggcaccacta cttcagagaa caaggccttt tcctctcctc gctccagtcc
     7621 caaaaacgaa ggggactaaa acgggtcttc gagaagacct tttttttgaa ttctgatgcg
     7681 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
     7741 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     7801 gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     7861 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     7921 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
     7981 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     8041 ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta
     8101 taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt
     8161 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
     8221 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     8281 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     8341 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     8401 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     8461 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     8521 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     8581 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     8641 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     8701 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     8761 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     8821 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     8881 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     8941 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     9001 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     9061 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     9121 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     9181 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     9241 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtgga
     9301 agccgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     9361 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     9421 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttag
//
