LOCUS       sequence-198453-       10715 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses negative-feedback dLwCas13a for mammalian RNA 
            targeting/imaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10715)
  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 10715)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            2017 Oct 4. doi: 10.1038/nature24049."
COMMENT     Sequence Label: pC015 - dLwCas13a-NF
FEATURES             Location/Qualifiers
     source          1..10715
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(281..1072,1073..1141))
                     /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     904..923
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1142..1246)
                     /gene="bla"
                     /label=AmpR promoter
     protein_bind    1297..1308
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     enhancer        1312..1691
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1694..1969
                     /label=chicken beta-actin promoter
     intron          1970..2981
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2905..2927
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2989..3008
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(3060..4478,4479..4481,4482..6194,6195..6197,
                     6198..6515)
                     /codon_start=1
                     /product="catalytically dead mutant of a Leptotrichia wadei
                     CRISPR-Cas effector that acts as an RNA-guided RNAse 
                     (Abudayyeh et al., 2017)"
                     /label=dLwCas13a
                     /note="contains the inactivating mutations R474A and 
                     R1046A"
                     /translation="MKVTKVDGISHKKYIEEGKLVKSTSEENRTSERLSELLSIRLDIY
                     IKNPDNASEEENRIRRENLKKFFSNKVLHLKDSVLYLKNRKEKNAVQDKNYSEEDISEY
                     DLKNKNSFSVLKKILLNEDVNSEELEIFRKDVEAKLNKINSLKYSFEENKANYQKINEN
                     NVEKVGGKSKRNIIYDYYRESAKRNDYINNVQEAFDKLYKKEDIEKLFFLIENSKKHEK
                     YKIREYYHKIIGRKNDKENFAKIIYEEIQNVNNIKELIEKIPDMSELKKSQVFYKYYLD
                     KEELNDKNIKYAFCHFVEIEMSQLLKNYVYKRLSNISNDKIKRIFEYQNLKKLIENKLL
                     NKLDTYVRNCGKYNYYLQVGEIATSDFIARNRQNEAFLRNIIGVSSVAYFSLRNILETE
                     NENGITGRMRGKTVKNNKGEEKYVSGEVDKIYNENKQNEVKENLKMFYSYDFNMDNKNE
                     IEDFFANIDEAISSIAHGIVHFNLELEGKDIFAFKNIAPSEISKKMFQNEINEKKLKLK
                     IFKQLNSANVFNYYEKDVIIKYLKNTKFNFVNKNIPFVPSFTKLYNKIEDLRNTLKFFW
                     SVPKDKEEKDAQIYLLKNIYYGEFLNKFVKNSKVFFKITNEVIKINKQRNQKTGHYKYQ
                     KFENIEKTVPVEYLAIIQSREMINNQDKEEKNTYIDFIQQIFLKGFIDYLNKNNLKYIE
                     SNNNNDNNDIFSKIKIKKDNKEKYDKILKNYEKHNRNKEIPHEINEFVREIKLGKILKY
                     TENLNMFYLILKLLNHKELTNLKGSLEKYQSANKEETFSDELELINLLNLDNNRVTEDF
                     ELEANEIGKFLDFNENKIKDRKELKKFDTNKIYFDGENIIKHRAFYNIKKYGMLNLLEK
                     IADKAKYKISLKELKEYSNKKNEIEKNYTMQQNLHRKYARPKKDEKFNDEDYKEYEKAI
                     GNIQKYTHLKNKVEFNELNLLQGLLLKILHRLVGYTSIWERDLRFRLKGEFPENHYIEE
                     IFNFDNSKNVKYKSGQIVEKYINFYKELYKDNVEKRSIYSDKKVKKLKQEKKDLYIANY
                     IAHFNYIPHAEISLLEVLENLRKLLSYDRKLKNAIMKSIVDILKEYGFVATFKIGADKK
                     IEIQTLESEKIVHLKNLKKKKLMTDRNSEELCELVKVMFEYKALE"
     CDS             join(6561..6563,6564..7271)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELY"
     CDS             join(6561..6563,6564..7271)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELY"
     primer_bind     complement(6603..6624)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6864..6883)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7211..7232
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             7317..7658
                     /codon_start=1
                     /product="zinc-finger domain from the CCR5-224 (+) 
                     zinc-finger nuclease that recognizes the left half-site 
                     target sequence in the human CCR5 gene (Perez et al., 2008;
                     Gross et al., 2013)"
                     /label=CCR5TC
                     /translation="FQCRICMRNFSDRSNLSRHIRTHTGEKPFACDICGRKFAISSNLN
                     SHTKIHTGSQKPFQCRICMRNFSRSDNLARHIRTHTGEKPFACDICGRKFATSGNLTRH
                     TKIHLRGSQL"
     CDS             7707..7832
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the rat zinc finger protein Kid-1 
                     (Witzgall et al., 1994)"
                     /label=KRAB-A
                     /translation="VSVTFEDVAVLFTRDEWKKLDLSQRSLYREVMLENYSNLASM"
     CDS             7842..7862
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     intron          7879..8011
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     7994..8013
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    8020..8608
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8073..8093)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8491..8502)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8635..8652)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8641..8752
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(8848..8868)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(8848..8866)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(8887..8903)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9009..9028)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    9074..9129
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(9128..9147)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(9490..9506)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    9514..9530
                     /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)."
     promoter        complement(join(9538..9544,9545..9562,9563..9568))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9583..9604
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9663..9858
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      9709..9844
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    9864..9998
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9901..9920)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9955..9974
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10066..10083)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(10237..10715,1..110))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10317..10336)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
       61 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      121 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      181 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      241 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      301 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
      361 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
      421 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
      481 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
      541 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
      601 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
      661 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
      721 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
      781 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
      841 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
      901 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
      961 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1021 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1081 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1141 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1201 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1261 aagtgccacc tgggtcgaca ttgattattg actagagtca tcctcatcgt cgacattgat
     1321 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     1381 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     1441 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     1501 gacgtcaatg ggtggactat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     1561 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     1621 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     1681 ctattaccat gggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc
     1741 tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg
     1801 gggggggggg gggcgcgcgc caggcggggc ggggcggggg cgagggcggg gcggggcgag
     1861 gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc
     1921 gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc
     1981 gttgccttcg ccccgtgccc cgctccgcgc cgcctcgcgc cgcccgcccc ggctctgact
     2041 gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta
     2101 gcgcttggtt taatgacggc tcgtttcttt tctgtggctg cgtgaaagcc ttaaagggct
     2161 ccgggagggc cctttgtgcg ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg
     2221 tggggagcgc cgcgtgcggc ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc
     2281 ggggctttgt gcgctccgcg tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg
     2341 tgcggggggg ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag
     2401 cagggggtgt gggcgcggcg gtcgggctgt aacccccccc tgcacccccc tccccgagtt
     2461 gctgagcacg gcccggcttc gggtgcgggg ctccgtgcgg ggcgtggcgc ggggctcgcc
     2521 gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc
     2581 ggggagggct cgggggaggg gcgcggcggc cccggagcgc cggcggctgt cgaggcgcgg
     2641 cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt
     2701 cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct agcgggcgcg
     2761 gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt cgtgcgtcgc
     2821 cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg gacggctgcc
     2881 ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagag
     2941 cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg
     3001 ttattgtgct gtctcatcat tttggcaaag aattcggtac cgcgggcccg ggcgccacca
     3061 tgaaagtgac caaggtcgac ggcatcagcc acaagaagta catcgaagag ggcaagctcg
     3121 tgaagtccac cagcgaggaa aaccggacca gcgagagact gagcgagctg ctgagcatcc
     3181 ggctggacat ctacatcaag aaccccgaca acgcctccga ggaagagaac cggatcagaa
     3241 gagagaacct gaagaagttc tttagcaaca aggtgctgca cctgaaggac agcgtgctgt
     3301 atctgaagaa ccggaaagaa aagaacgccg tgcaggacaa gaactatagc gaagaggaca
     3361 tcagcgagta cgacctgaaa aacaagaaca gcttctccgt gctgaagaag atcctgctga
     3421 acgaggacgt gaactctgag gaactggaaa tctttcggaa ggacgtggaa gccaagctga
     3481 acaagatcaa cagcctgaag tacagcttcg aagagaacaa ggccaactac cagaagatca
     3541 acgagaacaa cgtggaaaaa gtgggcggca agagcaagcg gaacatcatc tacgactact
     3601 acagagagag cgccaagcgc aacgactaca tcaacaacgt gcaggaagcc ttcgacaagc
     3661 tgtataagaa agaggatatc gagaaactgt ttttcctgat cgagaacagc aagaagcacg
     3721 agaagtacaa gatccgcgag tactatcaca agatcatcgg ccggaagaac gacaaagaga
     3781 acttcgccaa gattatctac gaagagatcc agaacgtgaa caacatcaaa gagctgattg
     3841 agaagatccc cgacatgtct gagctgaaga aaagccaggt gttctacaag tactacctgg
     3901 acaaagagga actgaacgac aagaatatta agtacgcctt ctgccacttc gtggaaatcg
     3961 agatgtccca gctgctgaaa aactacgtgt acaagcggct gagcaacatc agcaacgata
     4021 agatcaagcg gatcttcgag taccagaatc tgaaaaagct gatcgaaaac aaactgctga
     4081 acaagctgga cacctacgtg cggaactgcg gcaagtacaa ctactatctg caagtgggcg
     4141 agatcgccac ctccgacttt atcgcccgga accggcagaa cgaggccttc ctgagaaaca
     4201 tcatcggcgt gtccagcgtg gcctacttca gcctgaggaa catcctggaa accgagaacg
     4261 agaacggtat caccggccgg atgcggggca agaccgtgaa gaacaacaag ggcgaagaga
     4321 aatacgtgtc cggcgaggtg gacaagatct acaatgagaa caagcagaac gaagtgaaag
     4381 aaaatctgaa gatgttctac agctacgact tcaacatgga caacaagaac gagatcgagg
     4441 acttcttcgc caatatcgat gaggccatct ctagtattgc tcacggcatc gtgcatttca
     4501 acctggaact ggaaggcaag gacatcttcg ccttcaagaa tatcgccccc agcgagatct
     4561 ccaagaagat gtttcagaac gaaatcaacg aaaagaagct gaagctgaaa atcttcaagc
     4621 agctgaacag cgccaacgtg ttcaactact acgagaagga tgtgatcatc aagtacctga
     4681 agaataccaa gttcaacttc gtgaacaaaa acatcccctt cgtgcccagc ttcaccaagc
     4741 tgtacaacaa gattgaggac ctgcggaata ccctgaagtt tttttggagc gtgcccaagg
     4801 acaaagaaga gaaggacgcc cagatctacc tgctgaagaa tatctactac ggcgagttcc
     4861 tgaacaagtt cgtgaaaaac tccaaggtgt tctttaagat caccaatgaa gtgatcaaga
     4921 ttaacaagca gcggaaccag aaaaccggcc actacaagta tcagaagttc gagaacatcg
     4981 agaaaaccgt gcccgtggaa tacctggcca tcatccagag cagagagatg atcaacaacc
     5041 aggacaaaga ggaaaagaat acctacatcg actttattca gcagattttc ctgaagggct
     5101 tcatcgacta cctgaacaag aacaatctga agtatatcga gagcaacaac aacaatgaca
     5161 acaacgacat cttctccaag atcaagatca aaaaggataa caaagagaag tacgacaaga
     5221 tcctgaagaa ctatgagaag cacaatcgga acaaagaaat ccctcacgag atcaatgagt
     5281 tcgtgcgcga gatcaagctg gggaagattc tgaagtacac cgagaatctg aacatgtttt
     5341 acctgatcct gaagctgctg aaccacaaag agctgaccaa cctgaagggc agcctggaaa
     5401 agtaccagtc cgccaacaaa gaagaaacct tcagcgacga gttggaactg atcaacctgc
     5461 tgaacctgga caacaacaga gtgaccgagg acttcgagct ggaagccaac gagatcggca
     5521 agttcctgga cttcaacgaa aacaaaatca aggaccggaa agagctgaaa aagttcgaca
     5581 ccaacaagat ctatttcgac ggcgagaaca tcatcaagca ccgggccttc tacaatatca
     5641 agaaatacgg catgctgaat ctgctggaaa agatcgccga taaggccaag tataagatca
     5701 gcctgaaaga actgaaagag tacagcaaca agaagaatga gattgaaaag aactacacca
     5761 tgcagcagaa cctgcaccgg aagtacgcca gacccaagaa ggacgaaaag ttcaacgacg
     5821 aggactacaa agagtatgag aaggccatcg gcaacatcca gaagtacacc cacctgaaga
     5881 acaaggtgga attcaatgag ctgaacctgc tgcagggcct gctgctgaag atcctgcacc
     5941 ggctcgtggg ctacaccagc atctgggagc gggacctgag attccggctg aagggcgagt
     6001 ttcccgagaa ccactacatc gaggaaattt tcaatttcga caactccaag aatgtgaagt
     6061 acaaaagcgg ccagatcgtg gaaaagtata tcaacttcta caaagaactg tacaaggaca
     6121 atgtggaaaa gcggagcatc tactccgaca agaaagtgaa gaaactgaag caggagaaga
     6181 aagacctgta tattgcaaac tacatcgctc actttaatta tattcctcat gccgagatta
     6241 gcctgctgga agtgctggaa aacctgcgga agctgctgtc ctacgaccgg aagctgaaga
     6301 acgccatcat gaagtccatc gtggacattc tgaaagaata cggcttcgtg gccaccttca
     6361 agatcggcgc tgacaagaag atcgaaatcc agaccctgga atcagagaag atcgtgcacc
     6421 tgaagaatct gaagaaaaag aaactgatga ccgaccggaa cagcgaggaa ctgtgcgaac
     6481 tcgtgaaagt catgttcgag tacaaggccc tggaaggagg aggtggaagc ggaggaggag
     6541 gaagcggagg aggaggtagc gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     6601 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     6661 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     6721 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     6781 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     6841 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     6901 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     6961 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     7021 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     7081 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     7141 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     7201 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     7261 acgagctgta ctctagggga gctggcgctg gagcgggtgc aggggctggc tctagattcc
     7321 agtgccggat ctgcatgcgg aacttcagcg accggtccaa cctgagcagg cacatcagaa
     7381 cccacaccgg agaaaagccc ttcgcctgcg acatttgcgg ccggaagttc gccatcagca
     7441 gcaacctgaa cagccacacc aagatccaca ctggcagcca gaaacctttc cagtgcagaa
     7501 tttgtatgag aaactttagc agaagcgaca acctggccag acacatccgg acacatactg
     7561 gtgaaaaacc ttttgcctgt gatatctgtg gcagaaagtt tgccacctcc ggcaatctga
     7621 cccggcacac aaagattcac ctgcggggca gccagctatc gattgtcgac gctcctgaac
     7681 aacgtgaagg tgcttctcaa gtttctgttt ctgttacttt tgaagatgtt gctgttcttt
     7741 ttactcgtga tgaatggaaa aaacttgatc tttctcaacg ttctctttat cgtgaagtta
     7801 tgcttgaaaa ttattctaat cttgcttcta tggctggatc ccccaagaaa aagcgcaagg
     7861 tataactcga ggccgcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag
     7921 aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt
     7981 actgacatcc actttgcctt tctctccaca ggtgtcgaca atcaacctct ggattacaaa
     8041 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     8101 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     8161 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     8221 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     8281 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     8341 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     8401 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt
     8461 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     8521 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     8581 cggatctccc tttgggccgc ctccccgcct ggaattcgag ctcggtacga tcagcctcga
     8641 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     8701 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     8761 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     8821 gggaagacaa tagcccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta
     8881 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctaattcact cctcaggtgc
     8941 aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact
     9001 gagatcgatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat
     9061 ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg
     9121 tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat
     9181 ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta
     9241 taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa
     9301 gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca
     9361 tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc
     9421 ccagtcatag ctgtccctct tctcttatgg agatccctcg acctgcagcc caagcttggc
     9481 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     9541 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     9601 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat
     9661 ccgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     9721 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     9781 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     9841 ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa
     9901 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     9961 tgtccaaact catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca
    10021 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
    10081 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
    10141 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
    10201 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
    10261 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
    10321 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
    10381 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
    10441 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
    10501 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
    10561 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
    10621 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
    10681 agtatttggt atctgcgctc tgctgaagcc agtta
//
