LOCUS       sequence-373284-       14997 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 14997)
  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 14997)
  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: pJT242 Lenti pEF-dCas9-3XFLAG-ZNF10 KRAB[WSR7E...
FEATURES             Location/Qualifiers
     source          1..14997
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(64..83)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        255..634
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        636..834
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     789..809
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             852..1032
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1079..1204
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1697..1930
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2115..2159
                     /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    2457..2574
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2704..3882
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2935..3873
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3830..3850
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3898..3945
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(3946..3969,3970..3972,3973..6528,6529..6531,
                     6532..8046)
                     /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             8101..8121
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8137..8202
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             8254..8436
                     /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="TLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGYQ
                     LTKPDVILRLEKGEEP"
     CDS             8446..8499
                     /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             9226..9282
                     /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             9283..9681
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    9706..10294
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9759..9779)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10177..10188)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10297..10313)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10298..10314)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10819..10999
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(11025..11042)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11031..11255
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11301..11729
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11388..11407)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11598..11619
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11738..11758)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11743..12072
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      11923..12058
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     11985..12004
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12120..12167
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12186..12560
                     /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    12690..12811
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12727..12746)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12781..12800
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12860..12876)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12860..12876)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12873..12895)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12884..12900
                     /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(12908..12914,12915..12932,12933..12938))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12953..12974
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13091..13108)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13262..13850)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13342..13361)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(14021..14812,14813..14881))
                     /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     14644..14663
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14882..14986)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
       61 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
      121 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
      181 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
      241 cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
      301 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      361 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      421 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      481 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      541 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      601 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
      661 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      721 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      781 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt
      841 ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
      901 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
      961 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     1021 aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct
     1081 ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact
     1141 ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag
     1201 cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg
     1261 gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt
     1321 cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct
     1381 acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac
     1441 cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat
     1501 agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac
     1561 ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa
     1621 aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa
     1681 aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta
     1741 tgggcgcagc gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc
     1801 agcagcagaa caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag
     1861 tctggggcat caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc
     1921 aacagctcct ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt
     1981 ggaatgctag ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg
     2041 agtgggacag agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc
     2101 aaaaccagca agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt
     2161 ggaattggtt taacataaca aattggctgt ggtatataaa attattcata atgatagtag
     2221 gaggcttggt aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc
     2281 agggatattc accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc
     2341 ccgaaggaat agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga
     2401 acggatcggc actgcgtgcg ccaattctgc agacaaatgg cagtattcat ccacaatttt
     2461 aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca
     2521 acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt
     2581 tattacaggg acagcagaga tccagtttgg ttaattagct agctgcaaag atggataaag
     2641 ttttaaacag agaggaatct ttgcagctaa tggaccttct aggtcttgaa aggagtggga
     2701 attggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg
     2761 gggggagggg tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa
     2821 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt
     2881 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acaggtaagt
     2941 gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt atggcccttg cgtgccttga
     3001 attacttcca cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg
     3061 tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc
     3121 tggcctgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct
     3181 gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc
     3241 tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg
     3301 gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg
     3361 cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg
     3421 cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca
     3481 ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg
     3541 aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt
     3601 ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc
     3661 gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg
     3721 atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg gcacttgatg
     3781 taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag
     3841 acagtggttc aaagtttttt tcttccattt caggtgtcgt gacgtacggc caccatgaaa
     3901 aggccggcgg ccacgaaaaa ggccggccag gcaaaaaaga aaaaggacaa gaagtacagc
     3961 atcggcctgg ccatcggcac caactctgtg ggctgggccg tgatcaccga cgagtacaag
     4021 gtgcccagca agaaattcaa ggtgctgggc aacaccgacc ggcacagcat caagaagaac
     4081 ctgatcggag ccctgctgtt cgacagcggc gaaacagccg aggccacccg gctgaagaga
     4141 accgccagaa gaagatacac cagacggaag aaccggatct gctatctgca agagatcttc
     4201 agcaacgaga tggccaaggt ggacgacagc ttcttccaca gactggaaga gtccttcctg
     4261 gtggaagagg ataagaagca cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg
     4321 gcctaccacg agaagtaccc caccatctac cacctgagaa agaaactggt ggacagcacc
     4381 gacaaggccg acctgcggct gatctatctg gccctggccc acatgatcaa gttccggggc
     4441 cacttcctga tcgagggcga cctgaacccc gacaacagcg acgtggacaa gctgttcatc
     4501 cagctggtgc agacctacaa ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg
     4561 gacgccaagg ccatcctgtc tgccagactg agcaagagca gacggctgga aaatctgatc
     4621 gcccagctgc ccggcgagaa gaagaatggc ctgttcggca acctgattgc cctgagcctg
     4681 ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggatgccaa actgcagctg
     4741 agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg cgaccagtac
     4801 gccgacctgt ttctggccgc caagaacctg tccgacgcca tcctgctgag cgacatcctg
     4861 agagtgaaca ccgagatcac caaggccccc ctgagcgcct ctatgatcaa gagatacgac
     4921 gagcaccacc aggacctgac cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag
     4981 tacaaagaga ttttcttcga ccagagcaag aacggctacg ccggctacat tgacggcgga
     5041 gccagccagg aagagttcta caagttcatc aagcccatcc tggaaaagat ggacggcacc
     5101 gaggaactgc tcgtgaagct gaacagagag gacctgctgc ggaagcagcg gaccttcgac
     5161 aacggcagca tcccccacca gatccacctg ggagagctgc acgccattct gcggcggcag
     5221 gaagattttt acccattcct gaaggacaac cgggaaaaga tcgagaagat cctgaccttc
     5281 cgcatcccct actacgtggg ccctctggcc aggggaaaca gcagattcgc ctggatgacc
     5341 agaaagagcg aggaaaccat caccccctgg aacttcgagg aagtggtgga caagggcgct
     5401 tccgcccaga gcttcatcga gcggatgacc aacttcgata agaacctgcc caacgagaag
     5461 gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtataacga gctgaccaaa
     5521 gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc tgagcggcga gcagaaaaag
     5581 gccatcgtgg acctgctgtt caagaccaac cggaaagtga ccgtgaagca gctgaaagag
     5641 gactacttca agaaaatcga gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg
     5701 ttcaacgcct ccctgggcac ataccacgat ctgctgaaaa ttatcaagga caaggacttc
     5761 ctggacaatg aggaaaacga ggacattctg gaagatatcg tgctgaccct gacactgttt
     5821 gaggacagag agatgatcga ggaacggctg aaaacctatg cccacctgtt cgacgacaaa
     5881 gtgatgaagc agctgaagcg gcggagatac accggctggg gcaggctgag ccggaagctg
     5941 atcaacggca tccgggacaa gcagtccggc aagacaatcc tggatttcct gaagtccgac
     6001 ggcttcgcca acagaaactt catgcagctg atccacgacg acagcctgac ctttaaagag
     6061 gacatccaga aagcccaggt gtccggccag ggcgatagcc tgcacgagca cattgccaat
     6121 ctggccggca gccccgccat taagaagggc atcctgcaga cagtgaaggt ggtggacgag
     6181 ctcgtgaaag tgatgggccg gcacaagccc gagaacatcg tgatcgaaat ggccagagag
     6241 aaccagacca cccagaaggg acagaagaac agccgcgaga gaatgaagcg gatcgaagag
     6301 ggcatcaaag agctgggcag ccagatcctg aaagaacacc ccgtggaaaa cacccagctg
     6361 cagaacgaga agctgtacct gtactacctg cagaatgggc gggatatgta cgtggaccag
     6421 gaactggaca tcaaccggct gtccgactac gatgtggacc acatcgtgcc tcagagcttt
     6481 ctgaaggacg actccatcga caacaaggtg ctgaccagaa gcgacaaggc ccggggcaag
     6541 agcgacaacg tgccctccga agaggtcgtg aagaagatga agaactactg gcggcagctg
     6601 ctgaacgcca agctgattac ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc
     6661 ggcctgagcg aactggataa ggccggcttc atcaagagac agctggtgga aacccggcag
     6721 atcacaaagc acgtggcaca gatcctggac tcccggatga acactaagta cgacgagaat
     6781 gacaagctga tccgggaagt gaaagtgatc accctgaagt ccaagctggt gtccgatttc
     6841 cggaaggatt tccagtttta caaagtgcgc gagatcaaca actaccacca cgcccacgac
     6901 gcctacctga acgccgtcgt gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc
     6961 gagttcgtgt acggcgacta caaggtgtac gacgtgcgga agatgatcgc caagagcgag
     7021 caggaaatcg gcaaggctac cgccaagtac ttcttctaca gcaacatcat gaactttttc
     7081 aagaccgaga ttaccctggc caacggcgag atccggaagc ggcctctgat cgagacaaac
     7141 ggcgaaaccg gggagatcgt gtgggataag ggccgggatt ttgccaccgt gcggaaagtg
     7201 ctgagcatgc cccaagtgaa tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc
     7261 aaagagtcta tcctgcccaa gaggaacagc gataagctga tcgccagaaa gaaggactgg
     7321 gaccctaaga agtacggcgg cttcgacagc cccaccgtgg cctattctgt gctggtggtg
     7381 gccaaagtgg aaaagggcaa gtccaagaaa ctgaagagtg tgaaagagct gctggggatc
     7441 accatcatgg aaagaagcag cttcgagaag aatcccatcg actttctgga agccaagggc
     7501 tacaaagaag tgaaaaagga cctgatcatc aagctgccta agtactccct gttcgagctg
     7561 gaaaacggcc ggaagagaat gctggcctct gccggcgaac tgcagaaggg aaacgaactg
     7621 gccctgccct ccaaatatgt gaacttcctg tacctggcca gccactatga gaagctgaag
     7681 ggctcccccg aggataatga gcagaaacag ctgtttgtgg aacagcacaa gcactacctg
     7741 gacgagatca tcgagcagat cagcgagttc tccaagagag tgatcctggc cgacgctaat
     7801 ctggacaaag tgctgtccgc ctacaacaag caccgggata agcccatcag agagcaggcc
     7861 gagaatatca tccacctgtt taccctgacc aatctgggag cccctgccgc cttcaagtac
     7921 tttgacacca ccatcgaccg gaagaggtac accagcacca aagaggtgct ggacgccacc
     7981 ctgatccacc agagcatcac cggcctgtac gagacacgga tcgacctgtc tcagctggga
     8041 ggcgacagcg ctggaggagg tggaagcgga ggaggaggaa gcggaggagg aggtagcgga
     8101 cctaagaaaa agaggaaggt ggcggccgct ggatccgact acaaagacca tgacggtgat
     8161 tataaagatc atgacatcga ctacaaggat gacgatgaca agggtggatc cggatcaggc
     8221 tccgatgcca agtccttgac cgctgaggag gagaccctgg tcactttcaa ggacgttttt
     8281 gtggatttca cgagggaaga atggaaacta ctagacacag cacaacaaat agtatacaga
     8341 aacgtaatgc tagaaaacta caaaaaccta gtaagcctag gataccaact aacaaaacca
     8401 gacgtaatac taagactaga aaaaggagaa gaaccaggat ctggagaggg aaggggaagc
     8461 ctcctaactt gcggagatgt cgaggagaat cctggcccaa tggttagtaa gggagaggag
     8521 ttgataaaag agaatatgca catgaaactg tatatggaag ggacagtgga caaccatcac
     8581 ttcaagtgca cctctgaggg agaaggaaag ccatacgaag gcacccagac tatgcgaatt
     8641 aaggtcgtgg agggagggcc tctccccttc gccttcgata ttcttgcaac atctttcctc
     8701 tacgggagca aaaccttcat caatcacaca caaggcatac ctgacttttt taagcagagt
     8761 tttccagagg gctttacctg ggagcgtgtg actacatacg aagatggggg tgtgctgacg
     8821 gctacacagg atacttcact tcaggacggt tgtctgatct acaacgtcaa gatcaggggg
     8881 gtcaatttca cctcgaatgg accagtaatg caaaaaaaga ccctgggatg ggaagctttt
     8941 actgaaactc tctatcccgc cgacggcggc ctcgaaggca gaaacgatat ggctttgaag
     9001 ctggttggcg ggtcccatct aattgccaat gccaaaacca catataggag caaaaagcct
     9061 gcaaagaacc tgaagatgcc gggtgtgtac tatgttgact atagattaga gcggatcaaa
     9121 gaggcaaata acgaaacgta cgtggagcag cacgaggtag ccgtggcgcg ctactgcgat
     9181 ttaccctcca aactgggcca taaactgaac tgtacaggaa gcggagctac taacttcagc
     9241 ctgctgaagc aggctggcga cgtggaagaa aacccaggac ctatggccaa gcctttgtct
     9301 caagaagaat ccaccctcat tgaaagagca acggctacaa tcaacagcat ccccatctct
     9361 gaagactaca gcgtcgccag cgcagctctc tctagcgacg gccgcatctt cactggtgtc
     9421 aatgtatatc attttactgg gggaccttgt gcagaactcg tggtgctggg cactgctgct
     9481 gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc
     9541 ttgagcccct gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc
     9601 atagtgaagg acagtgatgg acagccgacg gcagttggga ttcgtgaatt gctgccctct
     9661 ggttatgtgt gggagggcta agaattcgat atcaagctta tcggtaatca acctctggat
     9721 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     9781 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     9841 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     9901 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     9961 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
    10021 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
    10081 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
    10141 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
    10201 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
    10261 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc tcgagaccta
    10321 gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga ttgtgcctgg
    10381 ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt acctttaaga
    10441 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa ggggggactg
    10501 gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat ctaccacaca
    10561 caaggctact tccctgattg gcagaactac acaccagggc cagggatcag atatccactg
    10621 acctttggat ggtgctacaa gctagtacca gttgagcaag agaaggtaga agaagccaat
    10681 gaaggagaga acacccgctt gttacaccct gtgagcctgc atgggatgga tgacccggag
    10741 agagaagtat tagagtggag gtttgacagc cgcctagcat ttcatcacat ggcccgagag
    10801 ctgcatccgg actgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
    10861 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
    10921 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
    10981 gtgtggaaaa tctctagcag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
    11041 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
    11101 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
    11161 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
    11221 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
    11281 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
    11341 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
    11401 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
    11461 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
    11521 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
    11581 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
    11641 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
    11701 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
    11761 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
    11821 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
    11881 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
    11941 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
    12001 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
    12061 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg
    12121 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact
    12181 aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag
    12241 cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg
    12301 ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc
    12361 cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt
    12421 cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg
    12481 agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg
    12541 tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg
    12601 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
    12661 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
    12721 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
    12781 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
    12841 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
    12901 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
    12961 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
    13021 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
    13081 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
    13141 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
    13201 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
    13261 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
    13321 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
    13381 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
    13441 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
    13501 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
    13561 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
    13621 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
    13681 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
    13741 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
    13801 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
    13861 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
    13921 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
    13981 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
    14041 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
    14101 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
    14161 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
    14221 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
    14281 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
    14341 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
    14401 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
    14461 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
    14521 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
    14581 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
    14641 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
    14701 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
    14761 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
    14821 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
    14881 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
    14941 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttcccc
//
