LOCUS       sequence-228361-       13585 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses catalytically inactive LwCas13a for mammalian RNA 
            targeting.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13585)
  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 13585)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 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: pC035 - dLwCas13a-msfGFP
FEATURES             Location/Qualifiers
     source          1..13585
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..580
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             598..778
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    825..950
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1443..1676
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1861..1905
                     /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    2203..2320
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2445..3623
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2676..3614
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3571..3591
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3630..3639
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3645..3665
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(4431..5849,5850..5852,5853..7565,7566..7568,
                     7569..7886)
                     /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             7902..7922
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7932..8012
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     misc_feature    8040..8628
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8093..8113)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8511..8522)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8631..8647)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             9153..9333
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(9359..9376)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9365..9589
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9635..10063
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9722..9741)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9932..9953
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10072..10092)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10077..10406
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10257..10392
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10319..10338
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10454..10501
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10520..10894
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    11024..11145
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11061..11080)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11115..11134
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11194..11210)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11194..11210)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11207..11229)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11218..11234
                     /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(11242..11248,11249..11266,11267..11272))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11287..11308
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11425..11442)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11596..12184)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11676..11695)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12355..13146,13147..13215))
                     /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     12978..12997
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13216..13320)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(13395..13414)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
      781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
      841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
      901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
      961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2341 cagagatcca gtttggttaa ttaatgcaaa gatggataaa gttttaaaca gagaggaatc
     2401 tttgcagcta atggaccttc taggtcttga aaggagtggg aattggctcc ggtgcccgtc
     2461 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt
     2521 gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc
     2581 tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg
     2641 ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg
     2701 ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc acctggctgc
     2761 agtacgtgat tcttgatccc gagcttcggg ttggaagtgg gtgggagagt tcgaggcctt
     2821 gcgcttaagg agccccttcg cctcgtgctt gagttgaggc ctggcctggg cgctggggcc
     2881 gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc tgctttcgat aagtctctag
     2941 ccatttaaaa tttttgatga cctgctgcga cgcttttttt ctggcaagat agtcttgtaa
     3001 atgcgggcca agatctgcac actggtattt cggtttttgg ggccgcgggc ggcgacgggg
     3061 cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa
     3121 tcggacgggg gtagtctcaa gctggccggc ctgctctggt gcctggcctc gcgccgccgt
     3181 gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc accagttgcg tgagcggaaa
     3241 gatggccgct tcccggccct gctgcaggga gctcaaaatg gaggacgcgg cgctcgggag
     3301 agcgggcggg tgagtcaccc acacaaagga aaagggcctt tccgtcctca gccgtcgctt
     3361 catgtgactc cacggagtac cgggcgccgt ccaggcacct cgattagttc tcgagctttt
     3421 ggagtacgtc gtctttaggt tggggggagg ggttttatgc gatggagttt ccccacactg
     3481 agtgggtgga gactgaagtt aggccagctt ggcacttgat gtaattctcc ttggaatttg
     3541 ccctttttga gtttggatct tggttcattc tcaagcctca gacagtggtt caaagttttt
     3601 ttcttccatt tcaggtgtcg tgacgtacgg ccaccatggc tagccccaaa aagaagagga
     3661 aagtgggatc cgtgagtaaa ggtgaagaac tcttcactgg agtagtgccc attctggtag
     3721 agcttgatgg agatgtaaat ggacataaat tctccgtcag gggcgaaggc gaaggggacg
     3781 ccacgaatgg taagctgact ctgaaattca tctgtacgac gggcaaactg cccgtcccat
     3841 ggcctacact cgtaacgacc ctcacctacg gcgtgcaatg cttttctcga tatcccgacc
     3901 acatgaaaca gcatgacttt ttcaagtctg caatgcctga aggttatgtt caagaaagga
     3961 ccatcagctt taaggatgat ggtacatata aaacccgagc cgaggttaaa tttgaagggg
     4021 acactctggt taatcgaatt gaactgaaag gtattgattt taaggaggac ggtaacatac
     4081 tggggcacaa gttggagtac aactttaaca gccataatgt gtatattacc gctgataagc
     4141 agaaaaatgg gataaaggcc aactttaaga tccgacataa tgtcgaagat ggtagtgttc
     4201 aactggctga tcattaccaa caaaatacgc ccatcggaga tggacctgta ctcttgcctg
     4261 acaatcatta tctctccacg caatcaaagc tttccaagga cccaaacgaa aagagagatc
     4321 acatggtcct tctggaattt gtgactgccg caggcatcac tctcggtatg gatgagctgt
     4381 acaagggagg aggtggaagc ggaggaggag gaagcggagg aggaggtagc atgaaagtga
     4441 ccaaggtcga cggcatcagc cacaagaagt acatcgaaga gggcaagctc gtgaagtcca
     4501 ccagcgagga aaaccggacc agcgagagac tgagcgagct gctgagcatc cggctggaca
     4561 tctacatcaa gaaccccgac aacgcctccg aggaagagaa ccggatcaga agagagaacc
     4621 tgaagaagtt ctttagcaac aaggtgctgc acctgaagga cagcgtgctg tatctgaaga
     4681 accggaaaga aaagaacgcc gtgcaggaca agaactatag cgaagaggac atcagcgagt
     4741 acgacctgaa aaacaagaac agcttctccg tgctgaagaa gatcctgctg aacgaggacg
     4801 tgaactctga ggaactggaa atctttcgga aggacgtgga agccaagctg aacaagatca
     4861 acagcctgaa gtacagcttc gaagagaaca aggccaacta ccagaagatc aacgagaaca
     4921 acgtggaaaa agtgggcggc aagagcaagc ggaacatcat ctacgactac tacagagaga
     4981 gcgccaagcg caacgactac atcaacaacg tgcaggaagc cttcgacaag ctgtataaga
     5041 aagaggatat cgagaaactg tttttcctga tcgagaacag caagaagcac gagaagtaca
     5101 agatccgcga gtactatcac aagatcatcg gccggaagaa cgacaaagag aacttcgcca
     5161 agattatcta cgaagagatc cagaacgtga acaacatcaa agagctgatt gagaagatcc
     5221 ccgacatgtc tgagctgaag aaaagccagg tgttctacaa gtactacctg gacaaagagg
     5281 aactgaacga caagaatatt aagtacgcct tctgccactt cgtggaaatc gagatgtccc
     5341 agctgctgaa aaactacgtg tacaagcggc tgagcaacat cagcaacgat aagatcaagc
     5401 ggatcttcga gtaccagaat ctgaaaaagc tgatcgaaaa caaactgctg aacaagctgg
     5461 acacctacgt gcggaactgc ggcaagtaca actactatct gcaagtgggc gagatcgcca
     5521 cctccgactt tatcgcccgg aaccggcaga acgaggcctt cctgagaaac atcatcggcg
     5581 tgtccagcgt ggcctacttc agcctgagga acatcctgga aaccgagaac gagaacggta
     5641 tcaccggccg gatgcggggc aagaccgtga agaacaacaa gggcgaagag aaatacgtgt
     5701 ccggcgaggt ggacaagatc tacaatgaga acaagcagaa cgaagtgaaa gaaaatctga
     5761 agatgttcta cagctacgac ttcaacatgg acaacaagaa cgagatcgag gacttcttcg
     5821 ccaatatcga tgaggccatc tctagtattg ctcacggcat cgtgcatttc aacctggaac
     5881 tggaaggcaa ggacatcttc gccttcaaga atatcgcccc cagcgagatc tccaagaaga
     5941 tgtttcagaa cgaaatcaac gaaaagaagc tgaagctgaa aatcttcaag cagctgaaca
     6001 gcgccaacgt gttcaactac tacgagaagg atgtgatcat caagtacctg aagaatacca
     6061 agttcaactt cgtgaacaaa aacatcccct tcgtgcccag cttcaccaag ctgtacaaca
     6121 agattgagga cctgcggaat accctgaagt ttttttggag cgtgcccaag gacaaagaag
     6181 agaaggacgc ccagatctac ctgctgaaga atatctacta cggcgagttc ctgaacaagt
     6241 tcgtgaaaaa ctccaaggtg ttctttaaga tcaccaatga agtgatcaag attaacaagc
     6301 agcggaacca gaaaaccggc cactacaagt atcagaagtt cgagaacatc gagaaaaccg
     6361 tgcccgtgga atacctggcc atcatccaga gcagagagat gatcaacaac caggacaaag
     6421 aggaaaagaa tacctacatc gactttattc agcagatttt cctgaagggc ttcatcgact
     6481 acctgaacaa gaacaatctg aagtatatcg agagcaacaa caacaatgac aacaacgaca
     6541 tcttctccaa gatcaagatc aaaaaggata acaaagagaa gtacgacaag atcctgaaga
     6601 actatgagaa gcacaatcgg aacaaagaaa tccctcacga gatcaatgag ttcgtgcgcg
     6661 agatcaagct ggggaagatt ctgaagtaca ccgagaatct gaacatgttt tacctgatcc
     6721 tgaagctgct gaaccacaaa gagctgacca acctgaaggg cagcctggaa aagtaccagt
     6781 ccgccaacaa agaagaaacc ttcagcgacg agttggaact gatcaacctg ctgaacctgg
     6841 acaacaacag agtgaccgag gacttcgagc tggaagccaa cgagatcggc aagttcctgg
     6901 acttcaacga aaacaaaatc aaggaccgga aagagctgaa aaagttcgac accaacaaga
     6961 tctatttcga cggcgagaac atcatcaagc accgggcctt ctacaatatc aagaaatacg
     7021 gcatgctgaa tctgctggaa aagatcgccg ataaggccaa gtataagatc agcctgaaag
     7081 aactgaaaga gtacagcaac aagaagaatg agattgaaaa gaactacacc atgcagcaga
     7141 acctgcaccg gaagtacgcc agacccaaga aggacgaaaa gttcaacgac gaggactaca
     7201 aagagtatga gaaggccatc ggcaacatcc agaagtacac ccacctgaag aacaaggtgg
     7261 aattcaatga gctgaacctg ctgcagggcc tgctgctgaa gatcctgcac cggctcgtgg
     7321 gctacaccag catctgggag cgggacctga gattccggct gaagggcgag tttcccgaga
     7381 accactacat cgaggaaatt ttcaatttcg acaactccaa gaatgtgaag tacaaaagcg
     7441 gccagatcgt ggaaaagtat atcaacttct acaaagaact gtacaaggac aatgtggaaa
     7501 agcggagcat ctactccgac aagaaagtga agaaactgaa gcaggagaag aaagacctgt
     7561 atattgcaaa ctacatcgct cactttaatt atattcctca tgccgagatt agcctgctgg
     7621 aagtgctgga aaacctgcgg aagctgctgt cctacgaccg gaagctgaag aacgccatca
     7681 tgaagtccat cgtggacatt ctgaaagaat acggcttcgt ggccaccttc aagatcggcg
     7741 ctgacaagaa gatcgaaatc cagaccctgg aatcagagaa gatcgtgcac ctgaagaatc
     7801 tgaagaaaaa gaaactgatg accgaccgga acagcgagga actgtgcgaa ctcgtgaaag
     7861 tcatgttcga gtacaaggcc ctggaaggat ccgagggagc tcccaagaaa aagcgcaagg
     7921 taggtagttc ctacccatac gatgttccag attacgctta tccctacgac gtgcctgatt
     7981 atgcataccc atatgatgtc cccgactatg cctaagaatt cgatatcaag cttatcgata
     8041 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     8101 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     8161 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     8221 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     8281 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     8341 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     8401 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     8461 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     8521 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     8581 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc
     8641 gacctcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca gctaccaatg
     8701 ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca gtcacacctc
     8761 aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
     8821 aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatatc cttgatctgt
     8881 ggatctacca cacacaaggc tacttccctg attggcagaa ctacacacca gggccaggga
     8941 tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag caagagaagg
     9001 tagaagaagc caatgaagga gagaacaccc gcttgttaca ccctgtgagc ctgcatggga
     9061 tggatgaccc ggagagagaa gtattagagt ggaggtttga cagccgccta gcatttcatc
     9121 acatggcccg agagctgcat ccggactgta ctgggtctct ctggttagac cagatctgag
     9181 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     9241 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     9301 gaccctttta gtcagtgtgg aaaatctcta gcagggcccg tttaaacccg ctgatcagcc
     9361 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     9421 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     9481 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     9541 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     9601 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     9661 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     9721 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     9781 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     9841 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     9901 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     9961 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
    10021 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
    10081 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
    10141 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
    10201 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
    10261 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
    10321 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
    10381 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
    10441 ctgatcagca cgtgttgaca attaatcatc ggcatagtat atcggcatag tataatacga
    10501 caaggtgagg aactaaacca tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg
    10561 cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttctccc gggacttcgt
    10621 ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca gcgcggtcca
    10681 ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct
    10741 gtacgccgag tggtcggagg tcgtgtccac gaacttccgg gacgcctccg ggccggccat
    10801 gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa
    10861 ctgcgtgcac ttcgtggccg aggagcagga ctgacacgtg ctacgagatt tcgattccac
    10921 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
    10981 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
    11041 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
    11101 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
    11161 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
    11221 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
    11281 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
    11341 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
    11401 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
    11461 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
    11521 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
    11581 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
    11641 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
    11701 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
    11761 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
    11821 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
    11881 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
    11941 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
    12001 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
    12061 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
    12121 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
    12181 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
    12241 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
    12301 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
    12361 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
    12421 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
    12481 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
    12541 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
    12601 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
    12661 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
    12721 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
    12781 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
    12841 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
    12901 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
    12961 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
    13021 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
    13081 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
    13141 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
    13201 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
    13261 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
    13321 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
    13381 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
    13441 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
    13501 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
    13561 gatgtacggg ccagatatac gcgtt
//
