LOCUS       sequence-369754-       15024 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 15024)
  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 15024)
  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: pJT350 Lenti
            pEF-dCas9-3XFLAG-MBDa-T2A-tagBFP-...
FEATURES             Location/Qualifiers
     source          1..15024
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    35..160
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    653..886
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1071..1115
                     /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    1413..1530
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1660..2838
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1891..2829
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2786..2806
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             2854..2901
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(2902..2925,2926..2928,2929..5484,5485..5487,
                     5488..7002)
                     /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             7057..7077
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7093..7158
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             7429..7482
                     /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             8209..8265
                     /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             8266..8664
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    8689..9277
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(8742..8762)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(9160..9171)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(9280..9296)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9281..9297)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             9802..9982
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(10008..10025)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    10014..10238
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      10284..10712
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10371..10390)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10581..10602
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(10721..10741)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        10726..11055
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10906..11041
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10968..10987
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        11103..11150
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             11169..11543
                     /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    11673..11794
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11710..11729)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     11764..11783
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11843..11859)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11843..11859)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11856..11878)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11867..11883
                     /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(11891..11897,11898..11915,11916..11921))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11936..11957
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12074..12091)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12245..12833)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12325..12344)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13004..13795,13796..13864))
                     /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     13627..13646
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13865..13969)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(14044..14063)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        14235..14614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        14616..14814
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     14769..14789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             14832..15012
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga ctcggcttgc
       61 tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa aaattttgac
      121 tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc gggggagaat
      181 tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat ataaattaaa
      241 acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg gcctgttaga
      301 aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc agacaggatc
      361 agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc atcaaaggat
      421 agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa acaaaagtaa
      481 gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat atgagggaca
      541 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
      601 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
      661 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
      721 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
      781 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
      841 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
      901 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
      961 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     1021 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     1081 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     1141 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     1201 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     1261 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg
     1321 gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa
     1381 ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg
     1441 tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta
     1501 caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca
     1561 gtttggttaa ttagctagct gcaaagatgg ataaagtttt aaacagagag gaatctttgc
     1621 agctaatgga ccttctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg
     1681 gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc
     1741 ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc
     1801 ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt
     1861 tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct
     1921 ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg gctgcagtac
     1981 gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct
     2041 taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc
     2101 gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt
     2161 taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg
     2221 ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt
     2281 gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga
     2341 cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc
     2401 gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg
     2461 ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg
     2521 gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt
     2581 gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt
     2641 acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg
     2701 gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt
     2761 tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt
     2821 ccatttcagg tgtcgtgacg tacggccacc atgaaaaggc cggcggccac gaaaaaggcc
     2881 ggccaggcaa aaaagaaaaa ggacaagaag tacagcatcg gcctggccat cggcaccaac
     2941 tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg
     3001 ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac
     3061 agcggcgaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga
     3121 cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac
     3181 gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag
     3241 cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc
     3301 atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc
     3361 tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg
     3421 aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag
     3481 ctgttcgagg aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc
     3541 agactgagca agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag
     3601 aatggcctgt tcggcaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc
     3661 aacttcgacc tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac
     3721 ctggacaacc tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag
     3781 aacctgtccg acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag
     3841 gcccccctga gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg
     3901 ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag
     3961 agcaagaacg gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag
     4021 ttcatcaagc ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac
     4081 agagaggacc tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc
     4141 cacctgggag agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag
     4201 gacaaccggg aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct
     4261 ctggccaggg gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc
     4321 ccctggaact tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg
     4381 atgaccaact tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg
     4441 tacgagtact tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg
     4501 agaaagcccg ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag
     4561 accaaccgga aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc
     4621 ttcgactccg tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac
     4681 cacgatctgc tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac
     4741 attctggaag atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa
     4801 cggctgaaaa cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg
     4861 agatacaccg gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag
     4921 tccggcaaga caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg
     4981 cagctgatcc acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc
     5041 ggccagggcg atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag
     5101 aagggcatcc tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac
     5161 aagcccgaga acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag
     5221 aagaacagcc gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag
     5281 atcctgaaag aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac
     5341 tacctgcaga atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc
     5401 gactacgatg tggaccacat cgtgcctcag agctttctga aggacgactc catcgacaac
     5461 aaggtgctga ccagaagcga caaggcccgg ggcaagagcg acaacgtgcc ctccgaagag
     5521 gtcgtgaaga agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag
     5581 agaaagttcg acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc
     5641 ggcttcatca agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc
     5701 ctggactccc ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa
     5761 gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa
     5821 gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga
     5881 accgccctga tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag
     5941 gtgtacgacg tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc
     6001 aagtacttct tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac
     6061 ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg
     6121 gataagggcc gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc
     6181 gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg
     6241 aacagcgata agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc
     6301 gacagcccca ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc
     6361 aagaaactga agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc
     6421 gagaagaatc ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg
     6481 atcatcaagc tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg
     6541 gcctctgccg gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac
     6601 ttcctgtacc tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag
     6661 aaacagctgt ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc
     6721 gagttctcca agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac
     6781 aacaagcacc gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc
     6841 ctgaccaatc tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag
     6901 aggtacacca gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc
     6961 ctgtacgaga cacggatcga cctgtctcag ctgggaggcg acagcgctgg aggaggtgga
     7021 agcggaggag gaggaagcgg aggaggaggt agcggaccta agaaaaagag gaaggtggcg
     7081 gccgctggat ccgactacaa agaccatgac ggtgattata aagatcatga catcgactac
     7141 aaggatgacg atgacaaggg tggatccgga tcaggctccc tgcagaagaa caagcagaga
     7201 ctgagaaacg accccctgaa ccagaacaag ggcaagcccg acctgaacac caccctgccc
     7261 atcagacaga ccgccagcat cttcaagcag cccgtgacca aggtgaccaa ccaccccagc
     7321 aacaaggtga agagcgaccc ccagagaatg aacgagcagc ccagacagct gttctgggag
     7381 aagagactgc agggcctgag cgccagcgac gtgaccgagg gatctggaga gggaagggga
     7441 agcctcctaa cttgcggaga tgtcgaggag aatcctggcc caatggttag taagggagag
     7501 gagttgataa aagagaatat gcacatgaaa ctgtatatgg aagggacagt ggacaaccat
     7561 cacttcaagt gcacctctga gggagaagga aagccatacg aaggcaccca gactatgcga
     7621 attaaggtcg tggagggagg gcctctcccc ttcgccttcg atattcttgc aacatctttc
     7681 ctctacggga gcaaaacctt catcaatcac acacaaggca tacctgactt ttttaagcag
     7741 agttttccag agggctttac ctgggagcgt gtgactacat acgaagatgg gggtgtgctg
     7801 acggctacac aggatacttc acttcaggac ggttgtctga tctacaacgt caagatcagg
     7861 ggggtcaatt tcacctcgaa tggaccagta atgcaaaaaa agaccctggg atgggaagct
     7921 tttactgaaa ctctctatcc cgccgacggc ggcctcgaag gcagaaacga tatggctttg
     7981 aagctggttg gcgggtccca tctaattgcc aatgccaaaa ccacatatag gagcaaaaag
     8041 cctgcaaaga acctgaagat gccgggtgtg tactatgttg actatagatt agagcggatc
     8101 aaagaggcaa ataacgaaac gtacgtggag cagcacgagg tagccgtggc gcgctactgc
     8161 gatttaccct ccaaactggg ccataaactg aactgtacag gaagcggagc tactaacttc
     8221 agcctgctga agcaggctgg cgacgtggaa gaaaacccag gacctatggc caagcctttg
     8281 tctcaagaag aatccaccct cattgaaaga gcaacggcta caatcaacag catccccatc
     8341 tctgaagact acagcgtcgc cagcgcagct ctctctagcg acggccgcat cttcactggt
     8401 gtcaatgtat atcattttac tgggggacct tgtgcagaac tcgtggtgct gggcactgct
     8461 gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga tcggaaatga gaacaggggc
     8521 atcttgagcc cctgcggacg gtgccgacag gtgcttctcg atctgcatcc tgggatcaaa
     8581 gccatagtga aggacagtga tggacagccg acggcagttg ggattcgtga attgctgccc
     8641 tctggttatg tgtgggaggg ctaagaattc gatatcaagc ttatcggtaa tcaacctctg
     8701 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     8761 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     8821 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     8881 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     8941 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     9001 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     9061 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
     9121 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     9181 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     9241 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg acctcgagac
     9301 ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc tgattgtgcc
     9361 tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca ggtaccttta
     9421 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     9481 ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg gatctaccac
     9541 acacaaggct acttccctga ttggcagaac tacacaccag ggccagggat cagatatcca
     9601 ctgacctttg gatggtgcta caagctagta ccagttgagc aagagaaggt agaagaagcc
     9661 aatgaaggag agaacacccg cttgttacac cctgtgagcc tgcatgggat ggatgacccg
     9721 gagagagaag tattagagtg gaggtttgac agccgcctag catttcatca catggcccga
     9781 gagctgcatc cggactgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     9841 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     9901 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     9961 tcagtgtgga aaatctctag cagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
    10021 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
    10081 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
    10141 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
    10201 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
    10261 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
    10321 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
    10381 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
    10441 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
    10501 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
    10561 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
    10621 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
    10681 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
    10741 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
    10801 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
    10861 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
    10921 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
    10981 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
    11041 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac
    11101 gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga
    11161 actaaaccat ggccaagttg accagtgccg ttccggtgct caccgcgcgc gacgtcgccg
    11221 gagcggtcga gttctggacc gaccggctcg ggttctcccg ggacttcgtg gaggacgact
    11281 tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg
    11341 tgccggacaa caccctggcc tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt
    11401 ggtcggaggt cgtgtccacg aacttccggg acgcctccgg gccggccatg accgagatcg
    11461 gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact
    11521 tcgtggccga ggagcaggac tgacacgtgc tacgagattt cgattccacc gccgccttct
    11581 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
    11641 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
    11701 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
    11761 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
    11821 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
    11881 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
    11941 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
    12001 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
    12061 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
    12121 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
    12181 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
    12241 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
    12301 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
    12361 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
    12421 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
    12481 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
    12541 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
    12601 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
    12661 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
    12721 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
    12781 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
    12841 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
    12901 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
    12961 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
    13021 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
    13081 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
    13141 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
    13201 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
    13261 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
    13321 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
    13381 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
    13441 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
    13501 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
    13561 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
    13621 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
    13681 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
    13741 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
    13801 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
    13861 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
    13921 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
    13981 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
    14041 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
    14101 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
    14161 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
    14221 cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
    14281 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
    14341 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
    14401 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
    14461 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
    14521 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
    14581 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
    14641 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
    14701 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
    14761 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt
    14821 ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
    14881 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
    14941 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
    15001 aaatctctag cagtggcgcc cgaa
//
