LOCUS       sequence-396589-       12694 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EnAsCas12a with a deactivated DNAse domain (D908A), N' 5x tandem 
            ALFA-tags and C' NLS for CRISPRa.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12694)
  AUTHORS   Griffith AL, Zheng F, McGee AV, Miller NW, Szegletes ZM, Reint G, 
            Gademann F, Nwolah I, Hegde M, Liu YV, Goodale A, Doench JG
  TITLE     Optimization of Cas12a for multiplexed genome-scale transcriptional 
            activation.
  JOURNAL   Cell Genom. 2023 Sep 1;3(9):100387. doi: 10.1016/j.xgen.2023.100387.
            eCollection 2023 Sep 13.
  PUBMED    37719144
REFERENCE   2  (bases 1 to 12694)
  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: "Cell 
            Genom."; date: "2023-09-1"; volume: "3(9):100387. doi"; pages: " 
            10.1016/j.xgen.2023.100387. eCollection 2023 Sep 13"
COMMENT     Sequence Label: pRDA_763
FEATURES             Location/Qualifiers
     source          1..12694
                     /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        2640..2851
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2856..2865
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2901..2921
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2931..2969
                     /codon_start=1
                     /product="rationally designed epitope tag that forms a 
                     stable alpha-helix (Gotzke et al., 2019)"
                     /label=ALFA-tag
                     /note="It may be beneficial to flank the ALFA-tag sequence 
                     with proline residues to preserve the helical structure."
                     /translation="SRLEEELRRRLTE"
     CDS             2994..3032
                     /codon_start=1
                     /product="rationally designed epitope tag that forms a 
                     stable alpha-helix (Gotzke et al., 2019)"
                     /label=ALFA-tag
                     /note="It may be beneficial to flank the ALFA-tag sequence 
                     with proline residues to preserve the helical structure."
                     /translation="SRLEEELRRRLTE"
     CDS             3057..3095
                     /codon_start=1
                     /product="rationally designed epitope tag that forms a 
                     stable alpha-helix (Gotzke et al., 2019)"
                     /label=ALFA-tag
                     /note="It may be beneficial to flank the ALFA-tag sequence 
                     with proline residues to preserve the helical structure."
                     /translation="SRLEEELRRRLTE"
     CDS             3120..3158
                     /codon_start=1
                     /product="rationally designed epitope tag that forms a 
                     stable alpha-helix (Gotzke et al., 2019)"
                     /label=ALFA-tag
                     /note="It may be beneficial to flank the ALFA-tag sequence 
                     with proline residues to preserve the helical structure."
                     /translation="SRLEEELRRRLTE"
     CDS             3183..3221
                     /codon_start=1
                     /product="rationally designed epitope tag that forms a 
                     stable alpha-helix (Gotzke et al., 2019)"
                     /label=ALFA-tag
                     /note="It may be beneficial to flank the ALFA-tag sequence 
                     with proline residues to preserve the helical structure."
                     /translation="SRLEEELRRRLTE"
     CDS             3252..7169
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="TQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARNDH
                     YKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRNA
                     IHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFDK
                     FTTYFSGFYRNRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHFE
                     NVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVLN
                     LAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLLR
                     NENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTGK
                     ITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTLK
                     KQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKAR
                     NYATKKPYSVEKFKLNFQMPTLARGWDVNREKNNGAILFVKNGLYYLGIMPKQKGRYKA
                     LSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEPL
                     EITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSIDL
                     SSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKGH
                     HGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLKD
                     QKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFFH
                     VPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIARGERNLIYITVIDSTGKILEQRS
                     LNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAVV
                     VLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQF
                     TSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYDV
                     KTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIENH
                     RFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQMR
                     NSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESK
                     DLKLQNGISNQDWLAYIQELRN"
     regulatory      6889..6898
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7176..7223
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7230..7310
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             7317..7370
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             7377..7775
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    7786..8374
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7839..7859)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8257..8268)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             8446..8679
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(8705..8722)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8711..8935
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8981..9409
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9068..9087)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9278..9299
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9418..9438)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9423..9752
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9603..9738
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     9665..9684
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        9800..9847
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             9866..10240
                     /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    10370..10491
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10407..10426)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10461..10480
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10540..10556)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10540..10556)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10553..10575)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10564..10580
                     /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(10588..10594,10595..10612,10613..10618))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10633..10654
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10771..10788)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10942..11530)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11022..11041)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11701..12492,12493..12561))
                     /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     12324..12343
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(12562..12666)
                     /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 aggtaccggg cagaggtctc gcttttttcg agacaacacg
     2641 ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac
     2701 gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg
     2761 cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct
     2821 ttttcgcaac gggtttgccg ccagaacaca gaggagccac catggccaaa cggacagccg
     2881 acggaagcga gttcgagtca ccaaagaaga agcggaaagt cggcaccagc agcagactgg
     2941 aggaggagct gagaagaagg ctgaccgagc caccaagtgg agggtctcct gcctccagac
     3001 tcgaggaaga gctcagaagg agactcaccg aagggtcccc aacatctact gaagaatcta
     3061 ggctcgaaga ggaactcagg aggaggctca cagaaggcgg accttcctca ggagcccctt
     3121 cacggctgga agaagaactg cggaggcgcc tcactgagcc atcttccaca gggggaccag
     3181 ccagtcgcct cgaagaagag ctcaggcgga gactgacaga gggagggccg agctctggcg
     3241 cacccggcag cacacagttc gagggcttta ccaacctgta tcaggtgagc aagacactgc
     3301 ggtttgagct gatcccacag ggcaagaccc tgaagcacat ccaggagcag ggcttcatcg
     3361 aggaggacaa ggcccgcaat gatcactaca aggagctgaa gcccatcatc gatcggatct
     3421 acaagaccta tgccgaccag tgcctgcagc tggtgcagct ggattgggag aacctgagcg
     3481 ccgccatcga ctcctataga aaggagaaaa ccgaggagac aaggaacgcc ctgatcgagg
     3541 agcaggccac atatcgcaat gccatccacg actacttcat cggccggaca gacaacctga
     3601 ccgatgccat caataagaga cacgccgaga tctacaaggg cctgttcaag gccgagctgt
     3661 ttaatggcaa ggtgctgaag cagctgggca ccgtgaccac aaccgagcac gagaacgccc
     3721 tgctgcggag cttcgacaag tttacaacct acttctccgg cttttatcgg aacaggaaga
     3781 acgtgttcag cgccgaggat atcagcacag ccatcccaca ccgcatcgtg caggacaact
     3841 tccccaagtt taaggagaat tgtcacatct tcacacgcct gatcaccgcc gtgcccagcc
     3901 tgcgggagca ctttgagaac gtgaagaagg ccatcggcat cttcgtgagc acctccatcg
     3961 aggaggtgtt ttccttccct ttttataacc agctgctgac acagacccag atcgacctgt
     4021 ataaccagct gctgggagga atctctcggg aggcaggcac cgagaagatc aagggcctga
     4081 acgaggtgct gaatctggcc atccagaaga atgatgagac agcccacatc atcgcctccc
     4141 tgccacacag attcatcccc ctgtttaagc agatcctgtc cgataggaac accctgtctt
     4201 tcatcctgga ggagtttaag agcgacgagg aagtgatcca gtccttctgc aagtacaaga
     4261 cactgctgag aaacgagaac gtgctggaga cagccgaggc cctgtttaac gagctgaaca
     4321 gcatcgacct gacacacatc ttcatcagcc acaagaagct ggagacaatc agcagcgccc
     4381 tgtgcgacca ctgggataca ctgaggaatg ccctgtatga gcggagaatc tccgagctga
     4441 caggcaagat caccaagtct gccaaggaga aggtgcagcg cagcctgaag cacgaggata
     4501 tcaacctgca ggagatcatc tctgccgcag gcaaggagct gagcgaggcc ttcaagcaga
     4561 aaaccagcga gatcctgtcc cacgcacacg ccgccctgga tcagccactg cctacaaccc
     4621 tgaagaagca ggaggagaag gagatcctga agtctcagct ggacagcctg ctgggcctgt
     4681 accacctgct ggactggttt gccgtggatg agtccaacga ggtggacccc gagttctctg
     4741 cccggctgac cggcatcaag ctggagatgg agccttctct gagcttctac aacaaggcca
     4801 gaaattatgc caccaagaag ccctactccg tggagaagtt caagctgaac tttcagatgc
     4861 ctacactggc ccgcggctgg gacgtgaatc gggagaagaa caatggcgcc atcctgtttg
     4921 tgaagaacgg cctgtactat ctgggcatca tgccaaagca gaagggcagg tataaggccc
     4981 tgagcttcga gcccacagag aaaaccagcg agggctttga taagatgtac tatgactact
     5041 tccctgatgc cgccaagatg atcccaaagt gcagcaccca gctgaaggcc gtgacagccc
     5101 actttcagac ccacacaacc cccatcctgc tgtccaacaa tttcatcgag cctctggaga
     5161 tcacaaagga gatctacgac ctgaacaatc ctgagaagga gccaaagaag tttcagacag
     5221 cctacgccaa gaaaaccggc gaccagaagg gctacagaga ggccctgtgc aagtggatcg
     5281 acttcacaag ggattttctg tccaagtata ccaagacaac ctctatcgat ctgtctagcc
     5341 tgcggccatc ctctcagtat aaggacctgg gcgagtacta tgccgagctg aatcccctgc
     5401 tgtaccacat cagcttccag agaatcgccg agaaggagat catggatgcc gtggagacag
     5461 gcaagctgta cctgttccag atctataaca aggactttgc caagggccac cacggcaagc
     5521 ctaatctgca cacactgtat tggaccggcc tgttttctcc agagaacctg gccaagacaa
     5581 gcatcaagct gaatggccag gccgagctgt tctaccgccc taagtccagg atgaagagga
     5641 tggcacaccg gctgggagag aagatgctga acaagaagct gaaggatcag aaaaccccaa
     5701 tccccgacac cctgtaccag gagctgtacg actatgtgaa tcacagactg tcccacgacc
     5761 tgtctgatga ggccagggcc ctgctgccca acgtgatcac caaggaggtg tctcacgaga
     5821 tcatcaagga taggcgcttt accagcgaca agttcttttt ccacgtgcct atcacactga
     5881 actatcaggc cgccaattcc ccatctaagt tcaaccagag ggtgaatgcc tacctgaagg
     5941 agcaccccga gacacctatc atcggcatcg cccggggcga gagaaacctg atctatatca
     6001 cagtgatcga ctccaccggc aagatcctgg agcagcggag cctgaacacc atccagcagt
     6061 ttgattacca gaagaagctg gacaacaggg agaaggagag ggtggcagca aggcaggcct
     6121 ggtctgtggt gggcacaatc aaggatctga agcagggcta tctgagccag gtcatccacg
     6181 agatcgtgga cctgatgatc cactaccagg ccgtggtggt gctggagaac ctgaatttcg
     6241 gctttaagag caagaggacc ggcatcgccg agaaggccgt gtaccagcag ttcgagaaga
     6301 tgctgatcga taagctgaat tgcctggtgc tgaaggacta tccagcagag aaagtgggag
     6361 gcgtgctgaa cccataccag ctgacagacc agttcacctc ctttgccaag atgggcaccc
     6421 agtctggctt cctgttttac gtgcctgccc catatacatc taagatcgat cccctgaccg
     6481 gcttcgtgga ccccttcgtg tggaaaacca tcaagaatca cgagagccgc aagcacttcc
     6541 tggagggctt cgactttctg cactacgacg tgaaaaccgg cgacttcatc ctgcacttta
     6601 agatgaacag aaatctgtcc ttccagaggg gcctgcccgg ctttatgcct gcatgggata
     6661 tcgtgttcga gaagaacgag acacagtttg acgccaaggg cacccctttc atcgccggca
     6721 agagaatcgt gccagtgatc gagaatcaca gattcaccgg cagataccgg gacctgtatc
     6781 ctgccaacga gctgatcgcc ctgctggagg agaagggcat cgtgttcagg gatggctcca
     6841 acatcctgcc aaagctgctg gagaatgacg attctcacgc catcgacacc atggtggccc
     6901 tgatccgcag cgtgctgcag atgcggaact ccaatgccgc cacaggcgag gactatatca
     6961 acagccccgt gcgcgatctg aatggcgtgt gcttcgactc ccggtttcag aacccagagt
     7021 ggcccatgga cgccgatgcc aatggcgcct accacatcgc cctgaagggc cagctgctgc
     7081 tgaatcacct gaaggagagc aaggatctga agctgcagaa cggcatctcc aatcaggact
     7141 ggctggccta catccaggag ctgcgcaacg ggtccaaaag gccggcggcc acgaaaaagg
     7201 ccggccaggc aaaaaagaaa aagggatcct acccatacga tgttccagat tacgcttatc
     7261 cctacgacgt gcctgattat gcatacccat atgatgtccc cgactatgcc ggaagcgagg
     7321 gcagaggaag ccttctaaca tgcggtgacg tggaggagaa tcccggccct tccgggatgg
     7381 ccaagccttt gtctcaagaa gaatccaccc tcattgaaag agcaacggct acaatcaaca
     7441 gcatccccat ctctgaggac tacagcgtcg ccagcgcagc tctctctagc gacggccgca
     7501 tcttcactgg tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc
     7561 tgggcactgc tgctgctgcg gcagctggca acctgacttg tatcgtcgcg atcggaaatg
     7621 agaacagggg catcttgagc ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc
     7681 ctgggatcaa agccatagtg aaggacagtg atggacagcc gacggcagtt gggattcgtg
     7741 aattgctgcc ctctggttat gtgtgggagg gctaataaat tcgaaaatca acctctggat
     7801 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     7861 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     7921 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     7981 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     8041 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     8101 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     8161 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
     8221 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     8281 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcctcgcct tcgccctcag
     8341 acgagtcgga tctccctttg ggccgcctcc ccgcgtcgac tttaagacca atgacttaca
     8401 aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc
     8461 actcccaacg aagataagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga
     8521 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     8581 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     8641 ccctcagacc cttttagtca gtgtggaaaa tctctagcag ggcccgttta aacccgctga
     8701 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     8761 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     8821 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     8881 ggggaggatt gggaagataa tagcaggcat gctggggatg cggtgggctc tatggcttct
     8941 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     9001 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     9061 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     9121 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     9181 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     9241 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     9301 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     9361 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     9421 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     9481 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     9541 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     9601 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     9661 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     9721 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     9781 tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata
     9841 atacgacaag gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac
     9901 cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga
     9961 cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc
    10021 ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga
    10081 cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc
    10141 ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc
    10201 cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga
    10261 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
    10321 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
    10381 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
    10441 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
    10501 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
    10561 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
    10621 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
    10681 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
    10741 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
    10801 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
    10861 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
    10921 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
    10981 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
    11041 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
    11101 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
    11161 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
    11221 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
    11281 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
    11341 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
    11401 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
    11461 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
    11521 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
    11581 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
    11641 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
    11701 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
    11761 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
    11821 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
    11881 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
    11941 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
    12001 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
    12061 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
    12121 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
    12181 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
    12241 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
    12301 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
    12361 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
    12421 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
    12481 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
    12541 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
    12601 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
    12661 tccgcgcaca tttccccgaa aagtgccacc tgac
//
