LOCUS       sequence-387204-       12020 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dCas12a effector fusion for manipulating transcription.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12020)
  AUTHORS   Tycko J, Van MV, Aradhana, DelRosso N, Ye H, Yao D, Valbuena R, 
            Vaughan-Jackson A, Xu X, Ludwig C, Spees K, Liu K, Gu M, Khare V, 
            Mukund AX, Suzuki PH, Arana S, Zhang C, Du PP, Ornstein TS, Hess GT,
            Kamber RA, Qi LS, Khalil AS, Bintu L, Bassik MC
  TITLE     Development of compact transcriptional effectors using 
            high-throughput measurements in diverse contexts.
  JOURNAL   Nat Biotechnol. 2024 Nov 1. doi: 10.1038/s41587-024-02442-6.
  PUBMED    39487265
REFERENCE   2  (bases 1 to 12020)
  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: "Nat 
            Biotechnol. 2024 Nov 1. doi: 10.1038/s41587-024-02442-6."
COMMENT     Sequence Label: pAR012 Lenti pEF-dEnAsCas12a-NLS-ZNF10
            KRAB-3X...
FEATURES             Location/Qualifiers
     source          1..12020
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     100..118
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(156..178)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     278..297
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     491..513
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     505..522
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     506..522
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        537..763
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             764..944
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    991..1116
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1609..1842
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2027..2071
                     /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    2349..2464
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1 (lacking the first T)"
     promoter        2616..3793
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2847..3784
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3741..3761
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3818..3838
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3863..7783
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="MTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARND
                     HYKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRN
                     AIHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFD
                     KFTTYFSGFYRNRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHF
                     ENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVL
                     NLAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLL
                     RNENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTG
                     KITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTL
                     KKQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKA
                     RNYATKKPYSVEKFKLNFQMPTLARGWDVNREKNNGAILFVKNGLYYLGIMPKQKGRYK
                     ALSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEP
                     LEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSID
                     LSSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKG
                     HHGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLK
                     DQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFF
                     HVPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIARGERNLIYITVIDSTGKILEQR
                     SLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAV
                     VVLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQ
                     FTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYD
                     VKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIEN
                     HRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQM
                     RNSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKES
                     KDLKLQNGISNQDWLAYIQELRN"
     CDS             7784..7831
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7838..7918
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             7967..8152
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEP"
     CDS             8162..8215
                     /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             8222..8620
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8630..9218
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8683..8703)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9101..9112)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             9292..9525
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9597..9718
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9634..9653)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9688..9707
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      9758..9893
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(9926..9942)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9926..9942)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9939..9961)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9950..9966
                     /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(9974..9980,9981..9998,9999..10004))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10019..10040
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(10157..10174)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10328..10916)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(11087..11878,11879..11947))
                     /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     11710..11729
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(11948..12020,1..32))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
       61 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
      121 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
      181 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
      241 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
      301 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
      361 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
      421 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
      481 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct gacgcgtgta
      541 gtcttatgca atactcttgt agtcttgcaa catggtaacg atgagttagc aacatgcctt
      601 acaaggagag aaaaagcacc gtgcatgccg attggtggaa gtaaggtggt acgatcgtgc
      661 cttattagga aggcaacaga cgggtctgac atggattgga cgaaccactg aattgccgca
      721 ttgcagagat attgtattta agtgcctagc tcgatacata aacgggtctc tctggttaga
      781 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
      841 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
      901 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg
      961 gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa
     1021 gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc
     1081 ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga
     1141 tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat
     1201 atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca
     1261 tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa
     1321 gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag
     1381 ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc
     1441 accgcacagc aagcggccac tgatcttcag acctggagga ggagatatga gggacaattg
     1501 gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac
     1561 caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt
     1621 ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt
     1681 acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat
     1741 tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag
     1801 aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc
     1861 tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct
     1921 ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac
     1981 aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga
     2041 attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct
     2101 gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt
     2161 tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac
     2221 ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga
     2281 gagagacaga gacagatcca ttcgattagt gaacggatct cgacggtatc gttgcttgga
     2341 tcgagcaatt taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag
     2401 acataatagc aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa
     2461 atttctgcgt tgttgtcggt gctcgttctc tgctcttcac gctactgaat tcatcaccgg
     2521 ttcttcgaat ctagttgcaa agatggataa agttttaaac agagaggaat ctttgcagct
     2581 aatggacctt ctaggtcttg aaaggagtgg gaattggctc cggtgcccgt cagtgggcag
     2641 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg
     2701 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
     2761 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
     2821 gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc
     2881 tctttacggg ttatggccct tgcgtgcctt gaattacttc cactggctgc agtacgtgat
     2941 tcttgatccc gagcttcggg ttggaagtgg gtgggagagt tcgaggcctt gcgcttaagg
     3001 agccccttcg cctcgtgctt gagttgaggc ctggcctggg cgctggggcc gccgcgtgcg
     3061 aatctggtgg caccttcgcg cctgtctcgc tgctttcgat aagtctctag ccatttaaaa
     3121 tttttgatga cctgctgcga cgcttttttt ctggcaagat agtcttgtaa atgcgggcca
     3181 agatctgcac actggtattt cggtttttgg ggccgcgggc ggcgacgggg cccgtgcgtc
     3241 ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg ccaccgagaa tcggacgggg
     3301 gtagtctcaa gctggccggc ctgctctggt gcctggcctc gcgccgccgt gtatcgcccc
     3361 gccctgggcg gcaaggctgg cccggtcggc accagttgcg tgagcggaaa gatggccgct
     3421 tcccggccct gctgcaggga gctcaaaatg gaggacgcgg cgctcgggag agcgggcggg
     3481 tgagtcaccc acacaaagga aaagggcctt tccgtcctca gccgtcgctt catgtgactc
     3541 cacggagtac cgggcgccgt ccaggcacct cgattagttc tcgagctttt ggagtacgtc
     3601 gtctttaggt tggggggagg ggttttatgc gatggagttt ccccacactg agtgggtgga
     3661 gactgaagtt aggccagctt ggcacttgat gtaattctcc ttggaatttg ccctttttga
     3721 gtttggatct tggttcattc tcaagcctca gacagtggtt caaagttttt ttcttccatt
     3781 tcaggtgtcg tgagtgtcgt gagctagcgc caccatgcca aagaagaagc ggaaggtcgg
     3841 tatccacgga gtcccagcag ccatgacaca gttcgagggc tttaccaacc tgtatcaggt
     3901 gagcaagaca ctgcggtttg agctgatccc acagggcaag accctgaagc acatccagga
     3961 gcagggcttc atcgaggagg acaaggcccg caatgatcac tacaaggagc tgaagcccat
     4021 catcgatcgg atctacaaga cctatgccga ccagtgcctg cagctggtgc agctggattg
     4081 ggagaacctg agcgccgcca tcgactccta tagaaaggag aaaaccgagg agacaaggaa
     4141 cgccctgatc gaggagcagg ccacatatcg caatgccatc cacgactact tcatcggccg
     4201 gacagacaac ctgaccgatg ccatcaataa gagacacgcc gagatctaca agggcctgtt
     4261 caaggccgag ctgtttaatg gcaaggtgct gaagcagctg ggcaccgtga ccacaaccga
     4321 gcacgagaac gccctgctgc ggagcttcga caagtttaca acctacttct ccggctttta
     4381 tagaaacagg aagaacgtgt tcagcgccga ggatatcagc acagccatcc cacaccgcat
     4441 cgtgcaggac aacttcccca agtttaagga gaattgtcac atcttcacac gcctgatcac
     4501 cgccgtgccc agcctgcggg agcactttga gaacgtgaag aaggccatcg gcatcttcgt
     4561 gagcacctcc atcgaggagg tgttttcctt ccctttttat aaccagctgc tgacacagac
     4621 ccagatcgac ctgtataacc agctgctggg aggaatctct cgggaggcag gcaccgagaa
     4681 gatcaagggc ctgaacgagg tgctgaatct ggccatccag aagaatgatg agacagccca
     4741 catcatcgcc tccctgccac acagattcat ccccctgttt aagcagatcc tgtccgatag
     4801 gaacaccctg tctttcatcc tggaggagtt taagagcgac gaggaagtga tccagtcctt
     4861 ctgcaagtac aagacactgc tgagaaacga gaacgtgctg gagacagccg aggccctgtt
     4921 taacgagctg aacagcatcg acctgacaca catcttcatc agccacaaga agctggagac
     4981 aatcagcagc gccctgtgcg accactggga tacactgagg aatgccctgt atgagcggag
     5041 aatctccgag ctgacaggca agatcaccaa gtctgccaag gagaaggtgc agcgcagcct
     5101 gaagcacgag gatatcaacc tgcaggagat catctctgcc gcaggcaagg agctgagcga
     5161 ggccttcaag cagaaaacca gcgagatcct gtcccacgca cacgccgccc tggatcagcc
     5221 actgcctaca accctgaaga agcaggagga gaaggagatc ctgaagtctc agctggacag
     5281 cctgctgggc ctgtaccacc tgctggactg gtttgccgtg gatgagtcca acgaggtgga
     5341 ccccgagttc tctgcccggc tgaccggcat caagctggag atggagcctt ctctgagctt
     5401 ctacaacaag gccagaaatt atgccaccaa gaagccctac tccgtggaga agttcaagct
     5461 gaactttcag atgcctacac tggccagagg ctgggacgtg aatagagaga agaacaatgg
     5521 cgccatcctg tttgtgaaga acggcctgta ctatctgggc atcatgccaa agcagaaggg
     5581 caggtataag gccctgagct tcgagcccac agagaaaacc agcgagggct ttgataagat
     5641 gtactatgac tacttccctg atgccgccaa gatgatccca aagtgcagca cccagctgaa
     5701 ggccgtgaca gcccactttc agacccacac aacccccatc ctgctgtcca acaatttcat
     5761 cgagcctctg gagatcacaa aggagatcta cgacctgaac aatcctgaga aggagccaaa
     5821 gaagtttcag acagcctacg ccaagaaaac cggcgaccag aagggctaca gagaggccct
     5881 gtgcaagtgg atcgacttca caagggattt tctgtccaag tataccaaga caacctctat
     5941 cgatctgtct agcctgcggc catcctctca gtataaggac ctgggcgagt actatgccga
     6001 gctgaatccc ctgctgtacc acatcagctt ccagagaatc gccgagaagg agatcatgga
     6061 tgccgtggag acaggcaagc tgtacctgtt ccagatctat aacaaggact ttgccaaggg
     6121 ccaccacggc aagcctaatc tgcacacact gtattggacc ggcctgtttt ctccagagaa
     6181 cctggccaag acaagcatca agctgaatgg ccaggccgag ctgttctacc gccctaagtc
     6241 caggatgaag aggatggcac accggctggg agagaagatg ctgaacaaga agctgaagga
     6301 tcagaaaacc ccaatccccg acaccctgta ccaggagctg tacgactatg tgaatcacag
     6361 actgtcccac gacctgtctg atgaggccag ggccctgctg cccaacgtga tcaccaagga
     6421 ggtgtctcac gagatcatca aggataggcg ctttaccagc gacaagttct ttttccacgt
     6481 gcctatcaca ctgaactatc aggccgccaa ttccccatct aagttcaacc agagggtgaa
     6541 tgcctacctg aaggagcacc ccgagacacc tatcatcggc atcgcccggg gcgagagaaa
     6601 cctgatctat atcacagtga tcgactccac cggcaagatc ctggagcagc ggagcctgaa
     6661 caccatccag cagtttgatt accagaagaa gctggacaac agggagaagg agagggtggc
     6721 agcaaggcag gcctggtctg tggtgggcac aatcaaggat ctgaagcagg gctatctgag
     6781 ccaggtcatc cacgagatcg tggacctgat gatccactac caggccgtgg tggtgctgga
     6841 gaacctgaat ttcggcttta agagcaagag gaccggcatc gccgagaagg ccgtgtacca
     6901 gcagttcgag aagatgctga tcgataagct gaattgcctg gtgctgaagg actatccagc
     6961 agagaaagtg ggaggcgtgc tgaacccata ccagctgaca gaccagttca cctcctttgc
     7021 caagatgggc acccagtctg gcttcctgtt ttacgtgcct gccccatata catctaagat
     7081 cgatcccctg accggcttcg tggacccctt cgtgtggaaa accatcaaga atcacgagag
     7141 ccgcaagcac ttcctggagg gcttcgactt tctgcactac gacgtgaaaa ccggcgactt
     7201 catcctgcac tttaagatga acagaaatct gtccttccag aggggcctgc ccggctttat
     7261 gcctgcatgg gatatcgtgt tcgagaagaa cgagacacag tttgacgcca agggcacccc
     7321 tttcatcgcc ggcaagagaa tcgtgccagt gatcgagaat cacagattca ccggcagata
     7381 ccgggacctg tatcctgcca acgagctgat cgccctgctg gaggagaagg gcatcgtgtt
     7441 cagggatggc tccaacatcc tgccaaagct gctggagaat gacgattctc acgccatcga
     7501 cacgatggtg gccctgatcc gcagcgtgct gcagatgcgg aactccaatg ccgccacagg
     7561 cgaggactat atcaacagcc ccgtgcgcga tctgaatggc gtgtgcttcg actcccggtt
     7621 tcagaaccca gagtggccaa tggacgccga tgccaatggc gcctaccaca tcgccctgaa
     7681 gggccagctg ctgctgaatc acctgaagga gagcaaggat ctgaagctgc agaacggcat
     7741 ctccaatcag gactggctgg cctacatcca ggagctgcgc aacaaaaggc cggcggccac
     7801 gaaaaaggcc ggccaggcaa aaaagaaaaa gggatcctac ccatacgatg ttccagatta
     7861 cgcttatccc tacgacgtgc ctgattatgc atacccatat gatgtccccg actatgccgg
     7921 tggatccgga tcaggctccg atgctaagtc tctgactgcc tggtccagga cccttgtgac
     7981 cttcaaggac gtcttcgttg attttacgcg ggaagaatgg aaactactag acacagcaca
     8041 acaaatagta tacagaaacg taatgctaga aaactacaaa aacctagtaa gcctaggata
     8101 ccaactaaca aaaccagacg taatactaag actagaaaaa ggagaagaac caggatctgg
     8161 agagggcaga ggaagccttc taacatgcgg tgacgtggag gagaatcccg gcccttccgg
     8221 gatggccaag cctttgtctc aagaagaatc caccctcatt gaaagagcaa cggctacaat
     8281 caacagcatc cccatctctg aggactacag cgtcgccagc gcagctctct ctagcgacgg
     8341 ccgcatcttc actggtgtca atgtatatca ttttactggg ggaccttgtg cagaactcgt
     8401 ggtgctgggc actgctgctg ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg
     8461 aaatgagaac aggggcatct tgagcccctg cggacggtgc cgacaggtgc ttctcgatct
     8521 gcatcctggg atcaaagcca tagtgaagga cagtgatgga cagccgacgg cagttgggat
     8581 tcgtgaattg ctgccctctg gttatgtgtg ggagggctaa taagtcgaca atcaacctct
     8641 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     8701 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     8761 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     8821 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     8881 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     8941 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     9001 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     9061 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     9121 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtctcc gccttcgccc
     9181 tcagacgagt cggatctctc tttgggccgc ctccccgcct ggtaccttta agaccaatga
     9241 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
     9301 taattcactc ccaacgaaga taagatctgc tttttgcttg tactgggtct ctctggttag
     9361 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     9421 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     9481 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc
     9541 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact
     9601 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     9661 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     9721 atgtctggct ctagctatcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     9781 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     9841 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagactt
     9901 ttgcagagac caaattcgta atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc
     9961 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
    10021 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
    10081 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
    10141 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
    10201 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
    10261 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
    10321 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
    10381 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
    10441 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
    10501 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
    10561 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
    10621 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
    10681 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
    10741 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
    10801 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
    10861 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
    10921 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
    10981 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
    11041 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
    11101 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
    11161 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
    11221 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
    11281 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
    11341 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
    11401 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
    11461 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
    11521 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
    11581 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
    11641 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
    11701 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
    11761 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
    11821 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
    11881 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
    11941 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
    12001 gcggatacat atttgaatgt
//
