LOCUS       sequence-431411-       11290 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPRi, EFS-driven dCas9-KRAB (ZNF10) (Cas only).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11290)
  AUTHORS   McGee AV, Liu YV, Griffith AL, Szegletes ZM, Wen B, Kraus C, Miller 
            NW, Steger RJ, Escude Velasco B, Bosch JA, Zirin JD, Viswanatha R, 
            Sontheimer EJ, Goodale A, Greene MA, Green TM, Doench JG
  TITLE     Modular vector assembly enables rapid assessment of emerging CRISPR 
            technologies.
  JOURNAL   Cell Genom. 2024 Mar 13;4(3):100519. doi: 
            10.1016/j.xgen.2024.100519.
  PUBMED    38484704
REFERENCE   2  (bases 1 to 11290)
  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: "2024-03-13"; volume: "4(3):100519. doi"; pages: " 
            10.1016/j.xgen.2024.100519"
COMMENT     Sequence Label: pRDA_662
FEATURES             Location/Qualifiers
     source          1..11290
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        11..222
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      227..236
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             272..292
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(299..322,323..325,326..2881,2882..2884,2885..4399)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the N863A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKARGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             4430..4477
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             4517..4711
                     /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
                     QLTKPDVILRLEKGEEPWLV"
     CDS             4736..4789
                     /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             4796..5194
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    5205..5793
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5258..5278)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5676..5687)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5865..6098
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6176..6297
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6213..6232)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6267..6286
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6337..6472
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6492..6511)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6493..6511)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6521..6538)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6521..6537)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6530..6552)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6679..7134
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6766..6785)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6976..6997
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7160..7264
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7265..7333,7334..8125)
                     /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     complement(7483..7502)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8296..8884
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8785..8804
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9038..9055
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9172..9193
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9208..9213,9214..9231,9232..9238)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9246..9262
                     /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)."
     primer_bind     9251..9273
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9270..9286
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9270..9286
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9305..9325
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        9307..9325
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        9353..9579
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             9580..9760
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    9807..9932
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    10425..10658
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             10843..10887
                     /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    11121..11238
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 cccgggacac gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
       61 ggcaattgaa cgggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
      121 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
      181 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac agaggagcca ccatggccaa
      241 acggacagcc gacggaagcg agttcgagtc accaaagaag aagcggaaag tcggcagcga
      301 caagaagtac agcatcggcc tggccatcgg caccaactct gtgggctggg ccgtgatcac
      361 cgacgagtac aaggtgccca gcaagaaatt caaggtgctg ggcaacaccg accggcacag
      421 catcaagaag aacctgatcg gagccctgct gttcgacagc ggcgaaacag ccgaggccac
      481 ccggctgaag agaaccgcca gaagaagata caccagacgg aagaaccgga tctgctatct
      541 gcaagagatc ttcagcaacg agatggccaa ggtggacgac agcttcttcc acagactgga
      601 agagtccttc ctggtggaag aggataagaa gcacgagcgg caccccatct tcggcaacat
      661 cgtggacgag gtggcctacc acgagaagta ccccaccatc taccacctga gaaagaaact
      721 ggtggacagc accgacaagg ccgacctgcg gctgatctat ctggccctgg cccacatgat
      781 caagttccgg ggccacttcc tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga
      841 caagctgttc atccagctgg tgcagaccta caaccagctg ttcgaggaaa accccatcaa
      901 cgctagcggc gtggacgcca aggccatcct gtctgccaga ctgagcaaga gcagacggct
      961 ggaaaatctg atcgcccagc tgcccggcga gaagaagaat ggcctgttcg gcaacctgat
     1021 tgccctgagc ctgggcctga cccccaactt caagagcaac ttcgacctgg ccgaggatgc
     1081 caaactgcag ctgagcaagg acacctacga cgacgacctg gacaacctgc tggcccagat
     1141 cggcgaccag tacgccgacc tgtttctggc cgccaagaac ctgtccgacg ccatcctgct
     1201 gagcgacatc ctgagagtga acaccgagat caccaaggcc cccctgagcg cctctatgat
     1261 caagagatac gacgagcacc accaggacct gaccctgctg aaagctctcg tgcggcagca
     1321 gctgcctgag aagtacaaag agattttctt cgaccagagc aagaacggct acgccggcta
     1381 cattgacggc ggagccagcc aggaagagtt ctacaagttc atcaagccca tcctggaaaa
     1441 gatggacggc accgaggaac tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca
     1501 gcggaccttc gacaacggca gcatccccca ccagatccac ctgggagagc tgcacgccat
     1561 tctgcggcgg caggaagatt tttacccatt cctgaaggac aaccgggaaa agatcgagaa
     1621 gatcctgacc ttccgcatcc cctactacgt gggccctctg gccaggggaa acagcagatt
     1681 cgcctggatg accagaaaga gcgaggaaac catcaccccc tggaacttcg aggaagtggt
     1741 ggacaagggc gcttccgccc agagcttcat cgagcggatg accaacttcg ataagaacct
     1801 gcccaacgag aaggtgctgc ccaagcacag cctgctgtac gagtacttca ccgtgtataa
     1861 cgagctgacc aaagtgaaat acgtgaccga gggaatgaga aagcccgcct tcctgagcgg
     1921 cgagcagaaa aaggccatcg tggacctgct gttcaagacc aaccggaaag tgaccgtgaa
     1981 gcagctgaaa gaggactact tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg
     2041 cgtggaagat cggttcaacg cctccctggg cacataccac gatctgctga aaattatcaa
     2101 ggacaaggac ttcctggaca atgaggaaaa cgaggacatt ctggaagata tcgtgctgac
     2161 cctgacactg tttgaggaca gagagatgat cgaggaacgg ctgaaaacct atgcccacct
     2221 gttcgacgac aaagtgatga agcagctgaa gcggcggaga tacaccggct ggggcaggct
     2281 gagccggaag ctgatcaacg gcatccggga caagcagtcc ggcaagacaa tcctggattt
     2341 cctgaagtcc gacggcttcg ccaacagaaa cttcatgcag ctgatccacg acgacagcct
     2401 gacctttaaa gaggacatcc agaaagccca ggtgtccggc cagggcgata gcctgcacga
     2461 gcacattgcc aatctggccg gcagccccgc cattaagaag ggcatcctgc agacagtgaa
     2521 ggtggtggac gagctcgtga aagtgatggg ccggcacaag cccgagaaca tcgtgatcga
     2581 aatggccaga gagaaccaga ccacccagaa gggacagaag aacagccgcg agagaatgaa
     2641 gcggatcgaa gagggcatca aagagctggg cagccagatc ctgaaagaac acccagtgga
     2701 aaacacccag ctgcagaacg agaagctgta cctgtactac ctgcagaatg ggcgggatat
     2761 gtacgtggac caggaactgg acatcaaccg gctgtccgac tacgatgtgg accatatcgt
     2821 gcctcagagc tttctgaagg acgactccat cgacaacaag gtgctgacca gaagcgacaa
     2881 ggcccggggc aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta
     2941 ctggcggcag ctgctgaacg ccaagctgat tacccagaga aagttcgaca atctgaccaa
     3001 ggccgagaga ggcggcctga gcgaactgga taaggccggc ttcatcaaga gacagctggt
     3061 ggaaacccgg cagatcacaa agcacgtggc acagatcctg gactcccgga tgaacactaa
     3121 gtacgacgag aatgacaagc tgatccggga agtgaaagtg atcaccctga agtccaagct
     3181 ggtgtccgat ttccggaagg atttccagtt ttacaaagtg cgcgagatca acaactacca
     3241 ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc
     3301 taagctggaa agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat
     3361 cgccaagagc gagcaggaaa tcggcaaggc taccgccaag tacttcttct acagcaacat
     3421 catgaacttt ttcaagaccg agattaccct ggccaacggc gagatccgga agcggcctct
     3481 gatcgagaca aacggcgaaa ccggggagat cgtgtgggat aagggccggg attttgccac
     3541 cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac
     3601 aggcggcttc agcaaagagt ctatcctgcc caagaggaac agcgataagc tgatcgccag
     3661 aaagaaggac tgggacccta agaagtacgg cggcttcgac agccccaccg tggcctattc
     3721 tgtgctggtg gtggccaaag tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga
     3781 gctgctgggg atcaccatca tggaaagaag cagcttcgag aagaatccca tcgactttct
     3841 ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc
     3901 cctgttcgag ctggaaaacg gccggaagag aatgctggcc tctgccggcg aactgcagaa
     3961 gggaaacgaa ctggccctgc cctccaaata tgtgaacttc ctgtacctgg ccagccacta
     4021 tgagaagctg aagggctccc ccgaggataa tgagcagaaa cagctgtttg tggaacagca
     4081 caagcactac ctggacgaga tcatcgagca gatcagcgag ttctccaaga gagtgatcct
     4141 ggccgacgct aatctggaca aagtgctgtc cgcctacaac aagcaccggg ataagcccat
     4201 cagagagcag gccgagaata tcatccacct gtttaccctg accaatctgg gagcccctgc
     4261 cgccttcaag tactttgaca ccaccatcga ccggaagagg tacaccagca ccaaagaggt
     4321 gctggacgcc accctgatcc accagagcat caccggcctg tacgagacac gaatcgattt
     4381 gtctcagctg ggaggcgatg ggtccgggag cggcgggagc ggagggagca aaaggccggc
     4441 ggccacgaaa aaggccggcc aggcaaaaaa gaaaaagagc ggaggctccg acgccaagag
     4501 cctgaccgcc tggagcagaa ccctggtgac cttcaaggac gtgttcgtgg acttcaccag
     4561 agaggagtgg aagctgctgg acaccgccca gcagatcgtg tacagaaacg tgatgctgga
     4621 gaactacaag aacctggtga gcctgggcta ccagctgacc aagcccgacg tgatcctgag
     4681 actggagaag ggcgaggagc cctggctggt ggagagagag atccaccagg gaagcgaggg
     4741 cagaggaagc cttctaacat gcggtgacgt ggaggagaat cccggccctt ccgggatggc
     4801 caagcctttg tctcaagaag aatccaccct cattgaaaga gcaacggcta caatcaacag
     4861 catccccatc tctgaggact acagcgtcgc cagcgcagct ctctctagcg acggccgcat
     4921 cttcactggt gtcaatgtat atcattttac tgggggacct tgtgcagaac tcgtggtgct
     4981 gggcactgct gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga tcggaaatga
     5041 gaacaggggc atcttgagcc cctgcggacg gtgccgacag gtgcttctcg atctgcatcc
     5101 tgggatcaaa gccatagtga aggacagtga tggacagccg acggcagttg ggattcgtga
     5161 attgctgccc tctggttatg tgtgggaggg ctaataaatt cgaaaatcaa cctctggatt
     5221 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     5281 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     5341 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     5401 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     5461 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     5521 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     5581 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct
     5641 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     5701 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcctcgcctt cgccctcaga
     5761 cgagtcggat ctccctttgg gccgcctccc cgcgtcgact ttaagaccaa tgacttacaa
     5821 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
     5881 ctcccaacga agataagatc tgctttttgc ttgtactggg tctctctggt tagaccagat
     5941 ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt
     6001 gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc
     6061 cctcagaccc ttttagtcag tgtggaaaat ctctagcagt acgtatagta gttcatgtca
     6121 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt
     6181 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     6241 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     6301 tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     6361 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     6421 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagggacg
     6481 tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac
     6541 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     6601 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     6661 gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     6721 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     6781 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     6841 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     6901 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     6961 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     7021 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     7081 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg
     7141 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     7201 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     7261 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     7321 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     7381 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     7441 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     7501 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     7561 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     7621 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     7681 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     7741 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     7801 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     7861 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     7921 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     7981 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     8041 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     8101 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     8161 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     8221 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     8281 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     8341 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     8401 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     8461 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     8521 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     8581 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     8641 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     8701 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     8761 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     8821 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     8881 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     8941 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     9001 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     9061 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     9121 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     9181 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     9241 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     9301 gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctgcaagctt aatgtagtct
     9361 tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca tgccttacaa
     9421 ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga tcgtgcctta
     9481 ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt gccgcattgc
     9541 agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg gttagaccag
     9601 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
     9661 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
     9721 tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact
     9781 tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc
     9841 gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag
     9901 gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc
     9961 gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag
    10021 tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag
    10081 aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac
    10141 ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa
    10201 aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg
    10261 cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga
    10321 agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag
    10381 gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt
    10441 gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag
    10501 gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag
    10561 gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc
    10621 ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga
    10681 aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa
    10741 cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc
    10801 ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta
    10861 ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg
    10921 tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct
    10981 gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac
    11041 ctcccaaccc cgaggggacc ctggtaccca tgcatgcatg aattctagat cttgagacaa
    11101 atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg
    11161 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
    11221 acaaaaattc aaaattttcg ggtttattac agggacagca gagatccact ttggcgccgg
    11281 ctcgaggggg
//
