LOCUS       sequence-227471-       13585 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses active 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 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: pC014 - LwCas13a-msfGFP
FEATURES             Location/Qualifiers
     source          1..13585
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2682..3860
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2913..3851
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3808..3828
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3867..3876
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3882..3902
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3909..7364
                     /codon_start=1
                     /product="Leptotrichia wadei CRISPR-Cas effector that acts 
                     as an RNA-guided RNAse (Abudayyeh et al., 2017)"
                     /label=LwCas13a
                     /note="previously called C2c2"
                     /translation="MKVTKVDGISHKKYIEEGKLVKSTSEENRTSERLSELLSIRLDIY
                     IKNPDNASEEENRIRRENLKKFFSNKVLHLKDSVLYLKNRKEKNAVQDKNYSEEDISEY
                     DLKNKNSFSVLKKILLNEDVNSEELEIFRKDVEAKLNKINSLKYSFEENKANYQKINEN
                     NVEKVGGKSKRNIIYDYYRESAKRNDYINNVQEAFDKLYKKEDIEKLFFLIENSKKHEK
                     YKIREYYHKIIGRKNDKENFAKIIYEEIQNVNNIKELIEKIPDMSELKKSQVFYKYYLD
                     KEELNDKNIKYAFCHFVEIEMSQLLKNYVYKRLSNISNDKIKRIFEYQNLKKLIENKLL
                     NKLDTYVRNCGKYNYYLQVGEIATSDFIARNRQNEAFLRNIIGVSSVAYFSLRNILETE
                     NENGITGRMRGKTVKNNKGEEKYVSGEVDKIYNENKQNEVKENLKMFYSYDFNMDNKNE
                     IEDFFANIDEAISSIRHGIVHFNLELEGKDIFAFKNIAPSEISKKMFQNEINEKKLKLK
                     IFKQLNSANVFNYYEKDVIIKYLKNTKFNFVNKNIPFVPSFTKLYNKIEDLRNTLKFFW
                     SVPKDKEEKDAQIYLLKNIYYGEFLNKFVKNSKVFFKITNEVIKINKQRNQKTGHYKYQ
                     KFENIEKTVPVEYLAIIQSREMINNQDKEEKNTYIDFIQQIFLKGFIDYLNKNNLKYIE
                     SNNNNDNNDIFSKIKIKKDNKEKYDKILKNYEKHNRNKEIPHEINEFVREIKLGKILKY
                     TENLNMFYLILKLLNHKELTNLKGSLEKYQSANKEETFSDELELINLLNLDNNRVTEDF
                     ELEANEIGKFLDFNENKIKDRKELKKFDTNKIYFDGENIIKHRAFYNIKKYGMLNLLEK
                     IADKAKYKISLKELKEYSNKKNEIEKNYTMQQNLHRKYARPKKDEKFNDEDYKEYEKAI
                     GNIQKYTHLKNKVEFNELNLLQGLLLKILHRLVGYTSIWERDLRFRLKGEFPENHYIEE
                     IFNFDNSKNVKYKSGQIVEKYINFYKELYKDNVEKRSIYSDKKVKKLKQEKKDLYIRNY
                     IAHFNYIPHAEISLLEVLENLRKLLSYDRKLKNAIMKSIVDILKEYGFVATFKIGADKK
                     IEIQTLESEKIVHLKNLKKKKLMTDRNSEELCELVKVMFEYKALE"
     CDS             8139..8159
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8169..8249
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     misc_feature    8277..8865
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8330..8350)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8748..8759)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8868..8884)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             9390..9570
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(9596..9613)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9602..9826
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9872..10300
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9959..9978)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10169..10190
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10309..10329)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10314..10643
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10494..10629
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10556..10575
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10691..10738
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10757..11131
                     /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    11261..11382
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11298..11317)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11352..11371
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11431..11447)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11431..11447)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11444..11466)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11455..11471
                     /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(11479..11485,11486..11503,11504..11509))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11524..11545
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11662..11679)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11833..12421)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11913..11932)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12592..13383,13384..13452))
                     /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     13215..13234
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13453..13557)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta atgcaaagat ggataaagtt ttaaacagag aggaatcttt
     2641 gcagctaatg gaccttctag gtcttgaaag gagtgggaat tggctccggt gcccgtcagt
     2701 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa
     2761 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc
     2821 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc
     2881 tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc
     2941 ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc tggctgcagt
     3001 acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg aggccttgcg
     3061 cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc tggggccgcc
     3121 gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca
     3181 tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg
     3241 cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc
     3301 gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg
     3361 gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta
     3421 tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat
     3481 ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc tcgggagagc
     3541 gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat
     3601 gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg agcttttgga
     3661 gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc cacactgagt
     3721 gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc
     3781 tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc
     3841 ttccatttca ggtgtcgtga cgtacggcca ccatggctag ccccaaaaag aagaggaaag
     3901 tgggatccat gaaagtgacc aaggtcgacg gcatcagcca caagaagtac atcgaagagg
     3961 gcaagctcgt gaagtccacc agcgaggaaa accggaccag cgagagactg agcgagctgc
     4021 tgagcatccg gctggacatc tacatcaaga accccgacaa cgcctccgag gaagagaacc
     4081 ggatcagaag agagaacctg aagaagttct ttagcaacaa ggtgctgcac ctgaaggaca
     4141 gcgtgctgta tctgaagaac cggaaagaaa agaacgccgt gcaggacaag aactatagcg
     4201 aagaggacat cagcgagtac gacctgaaaa acaagaacag cttctccgtg ctgaagaaga
     4261 tcctgctgaa cgaggacgtg aactctgagg aactggaaat ctttcggaag gacgtggaag
     4321 ccaagctgaa caagatcaac agcctgaagt acagcttcga agagaacaag gccaactacc
     4381 agaagatcaa cgagaacaac gtggaaaaag tgggcggcaa gagcaagcgg aacatcatct
     4441 acgactacta cagagagagc gccaagcgca acgactacat caacaacgtg caggaagcct
     4501 tcgacaagct gtataagaaa gaggatatcg agaaactgtt tttcctgatc gagaacagca
     4561 agaagcacga gaagtacaag atccgcgagt actatcacaa gatcatcggc cggaagaacg
     4621 acaaagagaa cttcgccaag attatctacg aagagatcca gaacgtgaac aacatcaaag
     4681 agctgattga gaagatcccc gacatgtctg agctgaagaa aagccaggtg ttctacaagt
     4741 actacctgga caaagaggaa ctgaacgaca agaatattaa gtacgccttc tgccacttcg
     4801 tggaaatcga gatgtcccag ctgctgaaaa actacgtgta caagcggctg agcaacatca
     4861 gcaacgataa gatcaagcgg atcttcgagt accagaatct gaaaaagctg atcgaaaaca
     4921 aactgctgaa caagctggac acctacgtgc ggaactgcgg caagtacaac tactatctgc
     4981 aagtgggcga gatcgccacc tccgacttta tcgcccggaa ccggcagaac gaggccttcc
     5041 tgagaaacat catcggcgtg tccagcgtgg cctacttcag cctgaggaac atcctggaaa
     5101 ccgagaacga gaacggtatc accggccgga tgcggggcaa gaccgtgaag aacaacaagg
     5161 gcgaagagaa atacgtgtcc ggcgaggtgg acaagatcta caatgagaac aagcagaacg
     5221 aagtgaaaga aaatctgaag atgttctaca gctacgactt caacatggac aacaagaacg
     5281 agatcgagga cttcttcgcc aacatcgacg aggccatcag cagcatcaga cacggcatcg
     5341 tgcacttcaa cctggaactg gaaggcaagg acatcttcgc cttcaagaat atcgccccca
     5401 gcgagatctc caagaagatg tttcagaacg aaatcaacga aaagaagctg aagctgaaaa
     5461 tcttcaagca gctgaacagc gccaacgtgt tcaactacta cgagaaggat gtgatcatca
     5521 agtacctgaa gaataccaag ttcaacttcg tgaacaaaaa catccccttc gtgcccagct
     5581 tcaccaagct gtacaacaag attgaggacc tgcggaatac cctgaagttt ttttggagcg
     5641 tgcccaagga caaagaagag aaggacgccc agatctacct gctgaagaat atctactacg
     5701 gcgagttcct gaacaagttc gtgaaaaact ccaaggtgtt ctttaagatc accaatgaag
     5761 tgatcaagat taacaagcag cggaaccaga aaaccggcca ctacaagtat cagaagttcg
     5821 agaacatcga gaaaaccgtg cccgtggaat acctggccat catccagagc agagagatga
     5881 tcaacaacca ggacaaagag gaaaagaata cctacatcga ctttattcag cagattttcc
     5941 tgaagggctt catcgactac ctgaacaaga acaatctgaa gtatatcgag agcaacaaca
     6001 acaatgacaa caacgacatc ttctccaaga tcaagatcaa aaaggataac aaagagaagt
     6061 acgacaagat cctgaagaac tatgagaagc acaatcggaa caaagaaatc cctcacgaga
     6121 tcaatgagtt cgtgcgcgag atcaagctgg ggaagattct gaagtacacc gagaatctga
     6181 acatgtttta cctgatcctg aagctgctga accacaaaga gctgaccaac ctgaagggca
     6241 gcctggaaaa gtaccagtcc gccaacaaag aagaaacctt cagcgacgag ttggaactga
     6301 tcaacctgct gaacctggac aacaacagag tgaccgagga cttcgagctg gaagccaacg
     6361 agatcggcaa gttcctggac ttcaacgaaa acaaaatcaa ggaccggaaa gagctgaaaa
     6421 agttcgacac caacaagatc tatttcgacg gcgagaacat catcaagcac cgggccttct
     6481 acaatatcaa gaaatacggc atgctgaatc tgctggaaaa gatcgccgat aaggccaagt
     6541 ataagatcag cctgaaagaa ctgaaagagt acagcaacaa gaagaatgag attgaaaaga
     6601 actacaccat gcagcagaac ctgcaccgga agtacgccag acccaagaag gacgaaaagt
     6661 tcaacgacga ggactacaaa gagtatgaga aggccatcgg caacatccag aagtacaccc
     6721 acctgaagaa caaggtggaa ttcaatgagc tgaacctgct gcagggcctg ctgctgaaga
     6781 tcctgcaccg gctcgtgggc tacaccagca tctgggagcg ggacctgaga ttccggctga
     6841 agggcgagtt tcccgagaac cactacatcg aggaaatttt caatttcgac aactccaaga
     6901 atgtgaagta caaaagcggc cagatcgtgg aaaagtatat caacttctac aaagaactgt
     6961 acaaggacaa tgtggaaaag cggagcatct actccgacaa gaaagtgaag aaactgaagc
     7021 aggaaaaaaa ggacctgtac atccggaact acattgccca cttcaactac atcccccacg
     7081 ccgagattag cctgctggaa gtgctggaaa acctgcggaa gctgctgtcc tacgaccgga
     7141 agctgaagaa cgccatcatg aagtccatcg tggacattct gaaagaatac ggcttcgtgg
     7201 ccaccttcaa gatcggcgct gacaagaaga tcgaaatcca gaccctggaa tcagagaaga
     7261 tcgtgcacct gaagaatctg aagaaaaaga aactgatgac cgaccggaac agcgaggaac
     7321 tgtgcgaact cgtgaaagtc atgttcgagt acaaggccct ggaaggagga ggtggaagcg
     7381 gaggaggagg aagcggagga ggaggtagcg tgagtaaagg tgaagaactc ttcactggag
     7441 tagtgcccat tctggtagag cttgatggag atgtaaatgg acataaattc tccgtcaggg
     7501 gcgaaggcga aggggacgcc acgaatggta agctgactct gaaattcatc tgtacgacgg
     7561 gcaaactgcc cgtcccatgg cctacactcg taacgaccct cacctacggc gtgcaatgct
     7621 tttctcgata tcccgaccac atgaaacagc atgacttttt caagtctgca atgcctgaag
     7681 gttatgttca agaaaggacc atcagcttta aggatgatgg tacatataaa acccgagccg
     7741 aggttaaatt tgaaggggac actctggtta atcgaattga actgaaaggt attgatttta
     7801 aggaggacgg taacatactg gggcacaagt tggagtacaa ctttaacagc cataatgtgt
     7861 atattaccgc tgataagcag aaaaatggga taaaggccaa ctttaagatc cgacataatg
     7921 tcgaagatgg tagtgttcaa ctggctgatc attaccaaca aaatacgccc atcggagatg
     7981 gacctgtact cttgcctgac aatcattatc tctccacgca atcaaagctt tccaaggacc
     8041 caaacgaaaa gagagatcac atggtccttc tggaatttgt gactgccgca ggcatcactc
     8101 tcggtatgga tgagctgtac aagggatccg agggagctcc caagaaaaag cgcaaggtag
     8161 gtagttccta cccatacgat gttccagatt acgcttatcc ctacgacgtg cctgattatg
     8221 catacccata tgatgtcccc gactatgcct aagaattcga tatcaagctt atcgataatc
     8281 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     8341 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     8401 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     8461 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     8521 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     8581 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     8641 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     8701 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     8761 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     8821 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgtcgac
     8881 ctcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct accaatgctg
     8941 attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc acacctcagg
     9001 tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa
     9061 aggggggact ggaagggcta attcactccc aacgaagaca agatatcctt gatctgtgga
     9121 tctaccacac acaaggctac ttccctgatt ggcagaacta cacaccaggg ccagggatca
     9181 gatatccact gacctttgga tggtgctaca agctagtacc agttgagcaa gagaaggtag
     9241 aagaagccaa tgaaggagag aacacccgct tgttacaccc tgtgagcctg catgggatgg
     9301 atgacccgga gagagaagta ttagagtgga ggtttgacag ccgcctagca tttcatcaca
     9361 tggcccgaga gctgcatccg gactgtactg ggtctctctg gttagaccag atctgagcct
     9421 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
     9481 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
     9541 ccttttagtc agtgtggaaa atctctagca gggcccgttt aaacccgctg atcagcctcg
     9601 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     9661 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     9721 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     9781 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     9841 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     9901 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     9961 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
    10021 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
    10081 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
    10141 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
    10201 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
    10261 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
    10321 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
    10381 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
    10441 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
    10501 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
    10561 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
    10621 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
    10681 atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa
    10741 ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga
    10801 cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg acttcgtgga
    10861 ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg cggtccagga
    10921 ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta
    10981 cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc cggccatgac
    11041 cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg ccggcaactg
    11101 cgtgcacttc gtggccgagg agcaggactg acacgtgcta cgagatttcg attccaccgc
    11161 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
    11221 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
    11281 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
    11341 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
    11401 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
    11461 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
    11521 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
    11581 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
    11641 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
    11701 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
    11761 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
    11821 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
    11881 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
    11941 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
    12001 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
    12061 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
    12121 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
    12181 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
    12241 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
    12301 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
    12361 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
    12421 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
    12481 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
    12541 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
    12601 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
    12661 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
    12721 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
    12781 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
    12841 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
    12901 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
    12961 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
    13021 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
    13081 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
    13141 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
    13201 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
    13261 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
    13321 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
    13381 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
    13441 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
    13501 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
    13561 atttccccga aaagtgccac ctgac
//
