LOCUS       sequence-176600-       13584 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 13584)
  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 13584)
  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..13584
                     /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..3859
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2913..3850
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3807..3827
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3866..3875
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3881..3901
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(4667..6085,6086..6088,6089..7801,7802..7804,
                     7805..8122)
                     /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             8138..8158
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8168..8248
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     misc_feature    8276..8864
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8329..8349)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8747..8758)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8867..8883)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             9389..9569
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(9595..9612)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9601..9825
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9871..10299
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9958..9977)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10168..10189
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10308..10328)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10313..10642
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10493..10628
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10555..10574
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10690..10737
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10756..11130
                     /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    11260..11381
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11297..11316)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11351..11370
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11430..11446)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11430..11446)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11443..11465)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11454..11470
                     /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(11478..11484,11485..11502,11503..11508))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11523..11544
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11661..11678)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11832..12420)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11912..11931)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12591..13382,13383..13451))
                     /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     13214..13233
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13452..13556)
                     /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 tacttccact ggctgcagta
     3001 cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc
     3061 ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg
     3121 cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt ctctagccat
     3181 ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc
     3241 gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg acggggcccg
     3301 tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg
     3361 acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat
     3421 cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag cggaaagatg
     3481 gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct cgggagagcg
     3541 ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg
     3601 tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga gcttttggag
     3661 tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc acactgagtg
     3721 ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg aatttgccct
     3781 ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa gtttttttct
     3841 tccatttcag gtgtcgtgac gtacggccac catggctagc cccaaaaaga agaggaaagt
     3901 gggatccgtg agtaaaggtg aagaactctt cactggagta gtgcccattc tggtagagct
     3961 tgatggagat gtaaatggac ataaattctc cgtcaggggc gaaggcgaag gggacgccac
     4021 gaatggtaag ctgactctga aattcatctg tacgacgggc aaactgcccg tcccatggcc
     4081 tacactcgta acgaccctca cctacggcgt gcaatgcttt tctcgatatc ccgaccacat
     4141 gaaacagcat gactttttca agtctgcaat gcctgaaggt tatgttcaag aaaggaccat
     4201 cagctttaag gatgatggta catataaaac ccgagccgag gttaaatttg aaggggacac
     4261 tctggttaat cgaattgaac tgaaaggtat tgattttaag gaggacggta acatactggg
     4321 gcacaagttg gagtacaact ttaacagcca taatgtgtat attaccgctg ataagcagaa
     4381 aaatgggata aaggccaact ttaagatccg acataatgtc gaagatggta gtgttcaact
     4441 ggctgatcat taccaacaaa atacgcccat cggagatgga cctgtactct tgcctgacaa
     4501 tcattatctc tccacgcaat caaagctttc caaggaccca aacgaaaaga gagatcacat
     4561 ggtccttctg gaatttgtga ctgccgcagg catcactctc ggtatggatg agctgtacaa
     4621 gggaggaggt ggaagcggag gaggaggaag cggaggagga ggtagcatga aagtgaccaa
     4681 ggtcgacggc atcagccaca agaagtacat cgaagagggc aagctcgtga agtccaccag
     4741 cgaggaaaac cggaccagcg agagactgag cgagctgctg agcatccggc tggacatcta
     4801 catcaagaac cccgacaacg cctccgagga agagaaccgg atcagaagag agaacctgaa
     4861 gaagttcttt agcaacaagg tgctgcacct gaaggacagc gtgctgtatc tgaagaaccg
     4921 gaaagaaaag aacgccgtgc aggacaagaa ctatagcgaa gaggacatca gcgagtacga
     4981 cctgaaaaac aagaacagct tctccgtgct gaagaagatc ctgctgaacg aggacgtgaa
     5041 ctctgaggaa ctggaaatct ttcggaagga cgtggaagcc aagctgaaca agatcaacag
     5101 cctgaagtac agcttcgaag agaacaaggc caactaccag aagatcaacg agaacaacgt
     5161 ggaaaaagtg ggcggcaaga gcaagcggaa catcatctac gactactaca gagagagcgc
     5221 caagcgcaac gactacatca acaacgtgca ggaagccttc gacaagctgt ataagaaaga
     5281 ggatatcgag aaactgtttt tcctgatcga gaacagcaag aagcacgaga agtacaagat
     5341 ccgcgagtac tatcacaaga tcatcggccg gaagaacgac aaagagaact tcgccaagat
     5401 tatctacgaa gagatccaga acgtgaacaa catcaaagag ctgattgaga agatccccga
     5461 catgtctgag ctgaagaaaa gccaggtgtt ctacaagtac tacctggaca aagaggaact
     5521 gaacgacaag aatattaagt acgccttctg ccacttcgtg gaaatcgaga tgtcccagct
     5581 gctgaaaaac tacgtgtaca agcggctgag caacatcagc aacgataaga tcaagcggat
     5641 cttcgagtac cagaatctga aaaagctgat cgaaaacaaa ctgctgaaca agctggacac
     5701 ctacgtgcgg aactgcggca agtacaacta ctatctgcaa gtgggcgaga tcgccacctc
     5761 cgactttatc gcccggaacc ggcagaacga ggccttcctg agaaacatca tcggcgtgtc
     5821 cagcgtggcc tacttcagcc tgaggaacat cctggaaacc gagaacgaga acggtatcac
     5881 cggccggatg cggggcaaga ccgtgaagaa caacaagggc gaagagaaat acgtgtccgg
     5941 cgaggtggac aagatctaca atgagaacaa gcagaacgaa gtgaaagaaa atctgaagat
     6001 gttctacagc tacgacttca acatggacaa caagaacgag atcgaggact tcttcgccaa
     6061 tatcgatgag gccatctcta gtattgctca cggcatcgtg catttcaacc tggaactgga
     6121 aggcaaggac atcttcgcct tcaagaatat cgcccccagc gagatctcca agaagatgtt
     6181 tcagaacgaa atcaacgaaa agaagctgaa gctgaaaatc ttcaagcagc tgaacagcgc
     6241 caacgtgttc aactactacg agaaggatgt gatcatcaag tacctgaaga ataccaagtt
     6301 caacttcgtg aacaaaaaca tccccttcgt gcccagcttc accaagctgt acaacaagat
     6361 tgaggacctg cggaataccc tgaagttttt ttggagcgtg cccaaggaca aagaagagaa
     6421 ggacgcccag atctacctgc tgaagaatat ctactacggc gagttcctga acaagttcgt
     6481 gaaaaactcc aaggtgttct ttaagatcac caatgaagtg atcaagatta acaagcagcg
     6541 gaaccagaaa accggccact acaagtatca gaagttcgag aacatcgaga aaaccgtgcc
     6601 cgtggaatac ctggccatca tccagagcag agagatgatc aacaaccagg acaaagagga
     6661 aaagaatacc tacatcgact ttattcagca gattttcctg aagggcttca tcgactacct
     6721 gaacaagaac aatctgaagt atatcgagag caacaacaac aatgacaaca acgacatctt
     6781 ctccaagatc aagatcaaaa aggataacaa agagaagtac gacaagatcc tgaagaacta
     6841 tgagaagcac aatcggaaca aagaaatccc tcacgagatc aatgagttcg tgcgcgagat
     6901 caagctgggg aagattctga agtacaccga gaatctgaac atgttttacc tgatcctgaa
     6961 gctgctgaac cacaaagagc tgaccaacct gaagggcagc ctggaaaagt accagtccgc
     7021 caacaaagaa gaaaccttca gcgacgagtt ggaactgatc aacctgctga acctggacaa
     7081 caacagagtg accgaggact tcgagctgga agccaacgag atcggcaagt tcctggactt
     7141 caacgaaaac aaaatcaagg accggaaaga gctgaaaaag ttcgacacca acaagatcta
     7201 tttcgacggc gagaacatca tcaagcaccg ggccttctac aatatcaaga aatacggcat
     7261 gctgaatctg ctggaaaaga tcgccgataa ggccaagtat aagatcagcc tgaaagaact
     7321 gaaagagtac agcaacaaga agaatgagat tgaaaagaac tacaccatgc agcagaacct
     7381 gcaccggaag tacgccagac ccaagaagga cgaaaagttc aacgacgagg actacaaaga
     7441 gtatgagaag gccatcggca acatccagaa gtacacccac ctgaagaaca aggtggaatt
     7501 caatgagctg aacctgctgc agggcctgct gctgaagatc ctgcaccggc tcgtgggcta
     7561 caccagcatc tgggagcggg acctgagatt ccggctgaag ggcgagtttc ccgagaacca
     7621 ctacatcgag gaaattttca atttcgacaa ctccaagaat gtgaagtaca aaagcggcca
     7681 gatcgtggaa aagtatatca acttctacaa agaactgtac aaggacaatg tggaaaagcg
     7741 gagcatctac tccgacaaga aagtgaagaa actgaagcag gagaagaaag acctgtatat
     7801 tgcaaactac atcgctcact ttaattatat tcctcatgcc gagattagcc tgctggaagt
     7861 gctggaaaac ctgcggaagc tgctgtccta cgaccggaag ctgaagaacg ccatcatgaa
     7921 gtccatcgtg gacattctga aagaatacgg cttcgtggcc accttcaaga tcggcgctga
     7981 caagaagatc gaaatccaga ccctggaatc agagaagatc gtgcacctga agaatctgaa
     8041 gaaaaagaaa ctgatgaccg accggaacag cgaggaactg tgcgaactcg tgaaagtcat
     8101 gttcgagtac aaggccctgg aaggatccga gggagctccc aagaaaaagc gcaaggtagg
     8161 tagttcctac ccatacgatg ttccagatta cgcttatccc tacgacgtgc ctgattatgc
     8221 atacccatat gatgtccccg actatgccta agaattcgat atcaagctta tcgataatca
     8281 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     8341 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     8401 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     8461 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     8521 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     8581 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     8641 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     8701 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     8761 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     8821 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc
     8881 tcgagaccta gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga
     8941 ttgtgcctgg ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt
     9001 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     9061 ggggggactg gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat
     9121 ctaccacaca caaggctact tccctgattg gcagaactac acaccagggc cagggatcag
     9181 atatccactg acctttggat ggtgctacaa gctagtacca gttgagcaag agaaggtaga
     9241 agaagccaat gaaggagaga acacccgctt gttacaccct gtgagcctgc atgggatgga
     9301 tgacccggag agagaagtat tagagtggag gtttgacagc cgcctagcat ttcatcacat
     9361 ggcccgagag ctgcatccgg actgtactgg gtctctctgg ttagaccaga tctgagcctg
     9421 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt
     9481 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc
     9541 cttttagtca gtgtggaaaa tctctagcag ggcccgttta aacccgctga tcagcctcga
     9601 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     9661 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     9721 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     9781 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     9841 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     9901 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     9961 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
    10021 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
    10081 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
    10141 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
    10201 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
    10261 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
    10321 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
    10381 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
    10441 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
    10501 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
    10561 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
    10621 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
    10681 tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
    10741 gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac
    10801 gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag
    10861 gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac
    10921 caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac
    10981 gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc
    11041 gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc
    11101 gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc
    11161 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
    11221 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
    11281 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
    11341 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
    11401 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
    11461 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
    11521 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
    11581 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
    11641 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
    11701 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
    11761 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
    11821 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
    11881 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
    11941 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
    12001 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
    12061 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
    12121 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
    12181 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
    12241 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
    12301 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
    12361 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
    12421 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
    12481 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
    12541 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
    12601 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
    12661 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
    12721 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
    12781 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
    12841 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
    12901 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
    12961 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
    13021 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
    13081 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
    13141 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
    13201 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
    13261 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
    13321 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
    13381 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
    13441 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
    13501 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
    13561 tttccccgaa aagtgccacc tgac
//
