LOCUS       sequence-369770-       14928 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dCas9 effector fusion for manipulating transcription.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14928)
  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 14928)
  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: pJT254 Lenti
            pEF-dCas9-3XFLAG-NCOA3_TAD-T2A-ta...
FEATURES             Location/Qualifiers
     source          1..14928
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(50..69)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        241..620
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        622..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     775..795
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             838..1018
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1065..1190
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1683..1916
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2101..2145
                     /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    2443..2560
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2690..3868
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2921..3859
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3816..3836
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3884..3931
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(3932..3955,3956..3958,3959..6514,6515..6517,
                     6518..8032)
                     /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             8087..8107
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8123..8188
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             8363..8416
                     /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             9143..9199
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             9200..9598
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    9623..10211
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9676..9696)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10094..10105)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10214..10230)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10215..10231)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10736..10916
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10942..10959)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10948..11172
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11218..11646
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11305..11324)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11515..11536
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11655..11675)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11660..11989
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11840..11975
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11902..11921
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12037..12084
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12103..12477
                     /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    12607..12728
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12644..12663)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12698..12717
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12777..12793)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12777..12793)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12790..12812)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12801..12817
                     /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(12825..12831,12832..12849,12850..12855))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12870..12891
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13008..13025)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13179..13767)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13259..13278)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13938..14729,14730..14798))
                     /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     14561..14580
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14799..14903)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
       61 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
      121 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
      181 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
      241 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
      301 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      361 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      421 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      481 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      541 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      601 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      661 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      721 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      781 tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc ctgtactggg
      841 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
      901 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
      961 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     1021 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
     1081 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
     1141 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
     1201 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
     1261 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
     1321 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
     1381 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
     1441 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
     1501 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
     1561 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1621 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1681 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1741 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1801 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1861 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1921 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1981 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     2041 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     2101 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     2161 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     2221 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2281 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2341 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2401 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2461 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2521 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2581 cagagatcca gtttggttaa ttagctagct gcaaagatgg ataaagtttt aaacagagag
     2641 gaatctttgc agctaatgga ccttctaggt cttgaaagga gtgggaattg gctccggtgc
     2701 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
     2761 caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
     2821 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
     2881 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc
     2941 ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg
     3001 gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag
     3061 gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg
     3121 gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc
     3181 tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct
     3241 tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga
     3301 cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc
     3361 gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc
     3421 gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc
     3481 ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc
     3541 gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt
     3601 cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag
     3661 cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca
     3721 cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga
     3781 atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag
     3841 tttttttctt ccatttcagg tgtcgtgacg tacggccacc atgaaaaggc cggcggccac
     3901 gaaaaaggcc ggccaggcaa aaaagaaaaa ggacaagaag tacagcatcg gcctggccat
     3961 cggcaccaac tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa
     4021 attcaaggtg ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggagccct
     4081 gctgttcgac agcggcgaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag
     4141 atacaccaga cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc
     4201 caaggtggac gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa
     4261 gaagcacgag cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa
     4321 gtaccccacc atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct
     4381 gcggctgatc tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga
     4441 gggcgacctg aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac
     4501 ctacaaccag ctgttcgagg aaaaccccat caacgccagc ggcgtggacg ccaaggccat
     4561 cctgtctgcc agactgagca agagcagacg gctggaaaat ctgatcgccc agctgcccgg
     4621 cgagaagaag aatggcctgt tcggcaacct gattgccctg agcctgggcc tgacccccaa
     4681 cttcaagagc aacttcgacc tggccgagga tgccaaactg cagctgagca aggacaccta
     4741 cgacgacgac ctggacaacc tgctggccca gatcggcgac cagtacgccg acctgtttct
     4801 ggccgccaag aacctgtccg acgccatcct gctgagcgac atcctgagag tgaacaccga
     4861 gatcaccaag gcccccctga gcgcctctat gatcaagaga tacgacgagc accaccagga
     4921 cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca aagagatttt
     4981 cttcgaccag agcaagaacg gctacgccgg ctacattgac ggcggagcca gccaggaaga
     5041 gttctacaag ttcatcaagc ccatcctgga aaagatggac ggcaccgagg aactgctcgt
     5101 gaagctgaac agagaggacc tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc
     5161 ccaccagatc cacctgggag agctgcacgc cattctgcgg cggcaggaag atttttaccc
     5221 attcctgaag gacaaccggg aaaagatcga gaagatcctg accttccgca tcccctacta
     5281 cgtgggccct ctggccaggg gaaacagcag attcgcctgg atgaccagaa agagcgagga
     5341 aaccatcacc ccctggaact tcgaggaagt ggtggacaag ggcgcttccg cccagagctt
     5401 catcgagcgg atgaccaact tcgataagaa cctgcccaac gagaaggtgc tgcccaagca
     5461 cagcctgctg tacgagtact tcaccgtgta taacgagctg accaaagtga aatacgtgac
     5521 cgagggaatg agaaagcccg ccttcctgag cggcgagcag aaaaaggcca tcgtggacct
     5581 gctgttcaag accaaccgga aagtgaccgt gaagcagctg aaagaggact acttcaagaa
     5641 aatcgagtgc ttcgactccg tggaaatctc cggcgtggaa gatcggttca acgcctccct
     5701 gggcacatac cacgatctgc tgaaaattat caaggacaag gacttcctgg acaatgagga
     5761 aaacgaggac attctggaag atatcgtgct gaccctgaca ctgtttgagg acagagagat
     5821 gatcgaggaa cggctgaaaa cctatgccca cctgttcgac gacaaagtga tgaagcagct
     5881 gaagcggcgg agatacaccg gctggggcag gctgagccgg aagctgatca acggcatccg
     5941 ggacaagcag tccggcaaga caatcctgga tttcctgaag tccgacggct tcgccaacag
     6001 aaacttcatg cagctgatcc acgacgacag cctgaccttt aaagaggaca tccagaaagc
     6061 ccaggtgtcc ggccagggcg atagcctgca cgagcacatt gccaatctgg ccggcagccc
     6121 cgccattaag aagggcatcc tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat
     6181 gggccggcac aagcccgaga acatcgtgat cgaaatggcc agagagaacc agaccaccca
     6241 gaagggacag aagaacagcc gcgagagaat gaagcggatc gaagagggca tcaaagagct
     6301 gggcagccag atcctgaaag aacaccccgt ggaaaacacc cagctgcaga acgagaagct
     6361 gtacctgtac tacctgcaga atgggcggga tatgtacgtg gaccaggaac tggacatcaa
     6421 ccggctgtcc gactacgatg tggaccacat cgtgcctcag agctttctga aggacgactc
     6481 catcgacaac aaggtgctga ccagaagcga caaggcccgg ggcaagagcg acaacgtgcc
     6541 ctccgaagag gtcgtgaaga agatgaagaa ctactggcgg cagctgctga acgccaagct
     6601 gattacccag agaaagttcg acaatctgac caaggccgag agaggcggcc tgagcgaact
     6661 ggataaggcc ggcttcatca agagacagct ggtggaaacc cggcagatca caaagcacgt
     6721 ggcacagatc ctggactccc ggatgaacac taagtacgac gagaatgaca agctgatccg
     6781 ggaagtgaaa gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga aggatttcca
     6841 gttttacaaa gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct acctgaacgc
     6901 cgtcgtggga accgccctga tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg
     6961 cgactacaag gtgtacgacg tgcggaagat gatcgccaag agcgagcagg aaatcggcaa
     7021 ggctaccgcc aagtacttct tctacagcaa catcatgaac tttttcaaga ccgagattac
     7081 cctggccaac ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg aaaccgggga
     7141 gatcgtgtgg gataagggcc gggattttgc caccgtgcgg aaagtgctga gcatgcccca
     7201 agtgaatatc gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag agtctatcct
     7261 gcccaagagg aacagcgata agctgatcgc cagaaagaag gactgggacc ctaagaagta
     7321 cggcggcttc gacagcccca ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa
     7381 gggcaagtcc aagaaactga agagtgtgaa agagctgctg gggatcacca tcatggaaag
     7441 aagcagcttc gagaagaatc ccatcgactt tctggaagcc aagggctaca aagaagtgaa
     7501 aaaggacctg atcatcaagc tgcctaagta ctccctgttc gagctggaaa acggccggaa
     7561 gagaatgctg gcctctgccg gcgaactgca gaagggaaac gaactggccc tgccctccaa
     7621 atatgtgaac ttcctgtacc tggccagcca ctatgagaag ctgaagggct cccccgagga
     7681 taatgagcag aaacagctgt ttgtggaaca gcacaagcac tacctggacg agatcatcga
     7741 gcagatcagc gagttctcca agagagtgat cctggccgac gctaatctgg acaaagtgct
     7801 gtccgcctac aacaagcacc gggataagcc catcagagag caggccgaga atatcatcca
     7861 cctgtttacc ctgaccaatc tgggagcccc tgccgccttc aagtactttg acaccaccat
     7921 cgaccggaag aggtacacca gcaccaaaga ggtgctggac gccaccctga tccaccagag
     7981 catcaccggc ctgtacgaga cacggatcga cctgtctcag ctgggaggcg acagcgctgg
     8041 aggaggtgga agcggaggag gaggaagcgg aggaggaggt agcggaccta agaaaaagag
     8101 gaaggtggcg gccgctggat ccgactacaa agaccatgac ggtgattata aagatcatga
     8161 catcgactac aaggatgacg atgacaaggg tggatccgga tcaggctccg agggccagag
     8221 cgacgagaga gccctgctgg accagctgca caccctgctg agcaacaccg acgccaccgg
     8281 cctggaggag atcgacagag ccctgggcat ccccgagctg gtgaaccagg gccaggccct
     8341 ggagcccaag cagggatctg gagagggaag gggaagcctc ctaacttgcg gagatgtcga
     8401 ggagaatcct ggcccaatgg ttagtaaggg agaggagttg ataaaagaga atatgcacat
     8461 gaaactgtat atggaaggga cagtggacaa ccatcacttc aagtgcacct ctgagggaga
     8521 aggaaagcca tacgaaggca cccagactat gcgaattaag gtcgtggagg gagggcctct
     8581 ccccttcgcc ttcgatattc ttgcaacatc tttcctctac gggagcaaaa ccttcatcaa
     8641 tcacacacaa ggcatacctg acttttttaa gcagagtttt ccagagggct ttacctggga
     8701 gcgtgtgact acatacgaag atgggggtgt gctgacggct acacaggata cttcacttca
     8761 ggacggttgt ctgatctaca acgtcaagat caggggggtc aatttcacct cgaatggacc
     8821 agtaatgcaa aaaaagaccc tgggatggga agcttttact gaaactctct atcccgccga
     8881 cggcggcctc gaaggcagaa acgatatggc tttgaagctg gttggcgggt cccatctaat
     8941 tgccaatgcc aaaaccacat ataggagcaa aaagcctgca aagaacctga agatgccggg
     9001 tgtgtactat gttgactata gattagagcg gatcaaagag gcaaataacg aaacgtacgt
     9061 ggagcagcac gaggtagccg tggcgcgcta ctgcgattta ccctccaaac tgggccataa
     9121 actgaactgt acaggaagcg gagctactaa cttcagcctg ctgaagcagg ctggcgacgt
     9181 ggaagaaaac ccaggaccta tggccaagcc tttgtctcaa gaagaatcca ccctcattga
     9241 aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc
     9301 agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg
     9361 accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac
     9421 ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg
     9481 acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca
     9541 gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg agggctaaga
     9601 attcgatatc aagcttatcg gtaatcaacc tctggattac aaaatttgtg aaagattgac
     9661 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     9721 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     9781 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     9841 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     9901 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     9961 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
    10021 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
    10081 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
    10141 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
    10201 cgcctccccg catcgatacc gtcgacctcg agacctagaa aaacatggag caatcacaag
    10261 tagcaataca gcagctacca atgctgattg tgcctggcta gaagcacaag aggaggagga
    10321 ggtgggtttt ccagtcacac ctcaggtacc tttaagacca atgacttaca aggcagctgt
    10381 agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg
    10441 aagacaagat atccttgatc tgtggatcta ccacacacaa ggctacttcc ctgattggca
    10501 gaactacaca ccagggccag ggatcagata tccactgacc tttggatggt gctacaagct
    10561 agtaccagtt gagcaagaga aggtagaaga agccaatgaa ggagagaaca cccgcttgtt
    10621 acaccctgtg agcctgcatg ggatggatga cccggagaga gaagtattag agtggaggtt
    10681 tgacagccgc ctagcatttc atcacatggc ccgagagctg catccggact gtactgggtc
    10741 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
    10801 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
    10861 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagggc
    10921 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
    10981 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
    11041 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
    11101 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
    11161 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
    11221 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
    11281 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
    11341 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
    11401 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
    11461 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
    11521 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
    11581 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
    11641 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
    11701 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
    11761 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
    11821 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
    11881 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
    11941 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
    12001 agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag
    12061 tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt
    12121 gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg
    12181 ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg
    12241 accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg
    12301 tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc
    12361 cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc
    12421 gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac
    12481 gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
    12541 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
    12601 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
    12661 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
    12721 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
    12781 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
    12841 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
    12901 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
    12961 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
    13021 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
    13081 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
    13141 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
    13201 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
    13261 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
    13321 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
    13381 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
    13441 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
    13501 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
    13561 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
    13621 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
    13681 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
    13741 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
    13801 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
    13861 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
    13921 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
    13981 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
    14041 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
    14101 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
    14161 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
    14221 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
    14281 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
    14341 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
    14401 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
    14461 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
    14521 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
    14581 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
    14641 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
    14701 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
    14761 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
    14821 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
    14881 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacct
//
