LOCUS       sequence-369768-       15069 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 15069)
  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 15069)
  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: pJT256 Lenti
            pEF-dCas9-3XFLAG-N+F-T2A-tagBFP-P...
FEATURES             Location/Qualifiers
     source          1..15069
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(94..113)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        285..664
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        666..864
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     819..839
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             882..1062
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1109..1234
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1727..1960
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2145..2189
                     /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    2487..2604
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2734..3912
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2965..3903
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3860..3880
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3928..3975
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(3976..3999,4000..4002,4003..6558,6559..6561,
                     6562..8076)
                     /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             8131..8151
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8167..8232
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             8548..8601
                     /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             9328..9384
                     /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             9385..9783
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    9808..10396
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(9861..9881)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(10279..10290)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(10399..10415)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10400..10416)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             10921..11101
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(11127..11144)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11133..11357
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11403..11831
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11490..11509)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11700..11721
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(11840..11860)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        11845..12174
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      12025..12160
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     12087..12106
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        12222..12269
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             12288..12662
                     /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    12792..12913
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(12829..12848)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     12883..12902
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(12962..12978)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12962..12978)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12975..12997)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12986..13002
                     /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(13010..13016,13017..13034,13035..13040))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13055..13076
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13193..13210)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13364..13952)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13444..13463)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(14123..14914,14915..14983))
                     /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     14746..14765
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(14984..15069,1..19))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
       61 gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
      121 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
      181 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
      241 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac
      301 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      361 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      421 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      481 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      541 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      601 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      661 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      721 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      781 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      841 acggtgggag gtctatataa gcagcgcgtt ttgcctgtac tgggtctctc tggttagacc
      901 agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
      961 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
     1021 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
     1081 cttgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
     1141 gcgcacggca agaggcgagg ggcggcgact ggtgagtacg ccaaaaattt tgactagcgg
     1201 aggctagaag gagagagatg ggtgcgagag cgtcagtatt aagcggggga gaattagatc
     1261 gcgatgggaa aaaattcggt taaggccagg gggaaagaaa aaatataaat taaaacatat
     1321 agtatgggca agcagggagc tagaacgatt cgcagttaat cctggcctgt tagaaacatc
     1381 agaaggctgt agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga
     1441 acttagatca ttatataata cagtagcaac cctctattgt gtgcatcaaa ggatagagat
     1501 aaaagacacc aaggaagctt tagacaagat agaggaagag caaaacaaaa gtaagaccac
     1561 cgcacagcaa gcggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga
     1621 gaagtgaatt atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca
     1681 aggcaaagag aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc
     1741 ttgggttctt gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac
     1801 aggccagaca attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg
     1861 aggcgcaaca gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa
     1921 tcctggctgt ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg
     1981 gaaaactcat ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg
     2041 aacagatttg gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa
     2101 gcttaataca ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat
     2161 tattggaatt agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt
     2221 ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg
     2281 ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc
     2341 acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga
     2401 gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg ccaattctgc
     2461 agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg ggggtacagt
     2521 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     2581 caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga tccagtttgg
     2641 ttaattagct agctgcaaag atggataaag ttttaaacag agaggaatct ttgcagctaa
     2701 tggaccttct aggtcttgaa aggagtggga attggctccg gtgcccgtca gtgggcagag
     2761 cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg aaccggtgcc
     2821 tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt
     2881 cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc
     2941 aacgggtttg ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc
     3001 tttacgggtt atggcccttg cgtgccttga attacttcca cctggctgca gtacgtgatt
     3061 cttgatcccg agcttcgggt tggaagtggg tgggagagtt cgaggccttg cgcttaagga
     3121 gccccttcgc ctcgtgcttg agttgaggcc tggcctgggc gctggggccg ccgcgtgcga
     3181 atctggtggc accttcgcgc ctgtctcgct gctttcgata agtctctagc catttaaaat
     3241 ttttgatgac ctgctgcgac gctttttttc tggcaagata gtcttgtaaa tgcgggccaa
     3301 gatctgcaca ctggtatttc ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc
     3361 cagcgcacat gttcggcgag gcggggcctg cgagcgcggc caccgagaat cggacggggg
     3421 tagtctcaag ctggccggcc tgctctggtg cctggcctcg cgccgccgtg tatcgccccg
     3481 ccctgggcgg caaggctggc ccggtcggca ccagttgcgt gagcggaaag atggccgctt
     3541 cccggccctg ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt
     3601 gagtcaccca cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc
     3661 acggagtacc gggcgccgtc caggcacctc gattagttct cgagcttttg gagtacgtcg
     3721 tctttaggtt ggggggaggg gttttatgcg atggagtttc cccacactga gtgggtggag
     3781 actgaagtta ggccagcttg gcacttgatg taattctcct tggaatttgc cctttttgag
     3841 tttggatctt ggttcattct caagcctcag acagtggttc aaagtttttt tcttccattt
     3901 caggtgtcgt gacgtacggc caccatgaaa aggccggcgg ccacgaaaaa ggccggccag
     3961 gcaaaaaaga aaaaggacaa gaagtacagc atcggcctgg ccatcggcac caactctgtg
     4021 ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa ggtgctgggc
     4081 aacaccgacc ggcacagcat caagaagaac ctgatcggag ccctgctgtt cgacagcggc
     4141 gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac cagacggaag
     4201 aaccggatct gctatctgca agagatcttc agcaacgaga tggccaaggt ggacgacagc
     4261 ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca cgagcggcac
     4321 cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc caccatctac
     4381 cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct gatctatctg
     4441 gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga cctgaacccc
     4501 gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc
     4561 gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc tgccagactg
     4621 agcaagagca gacggctgga aaatctgatc gcccagctgc ccggcgagaa gaagaatggc
     4681 ctgttcggca acctgattgc cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc
     4741 gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga cgacctggac
     4801 aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc caagaacctg
     4861 tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac caaggccccc
     4921 ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac cctgctgaaa
     4981 gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga ccagagcaag
     5041 aacggctacg ccggctacat tgacggcgga gccagccagg aagagttcta caagttcatc
     5101 aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct gaacagagag
     5161 gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca gatccacctg
     5221 ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct gaaggacaac
     5281 cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg ccctctggcc
     5341 aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat caccccctgg
     5401 aacttcgagg aagtggtgga caagggcgct tccgcccaga gcttcatcga gcggatgacc
     5461 aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct gctgtacgag
     5521 tacttcaccg tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag
     5581 cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt caagaccaac
     5641 cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga gtgcttcgac
     5701 tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac ataccacgat
     5761 ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga ggacattctg
     5821 gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga ggaacggctg
     5881 aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg gcggagatac
     5941 accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa gcagtccggc
     6001 aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt catgcagctg
     6061 atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt gtccggccag
     6121 ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat taagaagggc
     6181 atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg gcacaagccc
     6241 gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg acagaagaac
     6301 agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag ccagatcctg
     6361 aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct gtactacctg
     6421 cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct gtccgactac
     6481 gatgtggacc acatcgtgcc tcagagcttt ctgaaggacg actccatcga caacaaggtg
     6541 ctgaccagaa gcgacaaggc ccggggcaag agcgacaacg tgccctccga agaggtcgtg
     6601 aagaagatga agaactactg gcggcagctg ctgaacgcca agctgattac ccagagaaag
     6661 ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa ggccggcttc
     6721 atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca gatcctggac
     6781 tcccggatga acactaagta cgacgagaat gacaagctga tccgggaagt gaaagtgatc
     6841 accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta caaagtgcgc
     6901 gagatcaaca actaccacca cgcccacgac gcctacctga acgccgtcgt gggaaccgcc
     6961 ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt acggcgacta caaggtgtac
     7021 gacgtgcgga agatgatcgc caagagcgag caggaaatcg gcaaggctac cgccaagtac
     7081 ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc caacggcgag
     7141 atccggaagc ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt gtgggataag
     7201 ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa tatcgtgaaa
     7261 aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa gaggaacagc
     7321 gataagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg cttcgacagc
     7381 cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa gtccaagaaa
     7441 ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag cttcgagaag
     7501 aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga cctgatcatc
     7561 aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat gctggcctct
     7621 gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt gaacttcctg
     7681 tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga gcagaaacag
     7741 ctgtttgtgg aacagcacaa gcactacctg gacgagatca tcgagcagat cagcgagttc
     7801 tccaagagag tgatcctggc cgacgctaat ctggacaaag tgctgtccgc ctacaacaag
     7861 caccgggata agcccatcag agagcaggcc gagaatatca tccacctgtt taccctgacc
     7921 aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg gaagaggtac
     7981 accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac cggcctgtac
     8041 gagacacgga tcgacctgtc tcagctggga ggcgacagcg ctggaggagg tggaagcgga
     8101 ggaggaggaa gcggaggagg aggtagcgga cctaagaaaa agaggaaggt ggcggccgct
     8161 ggatccgact acaaagacca tgacggtgat tataaagatc atgacatcga ctacaaggat
     8221 gacgatgaca agggtggatc cggatcaggc tccgagggcc agagcgacga gagagccctg
     8281 ctggaccagc tgcacaccct gctgagcaac accgacgcca ccggcctgga ggagatcgac
     8341 agagccctgg gcatccccga gctggtgaac cagggccagg ccctggagcc caagcaggga
     8401 ggaggaggta gcggacacga gaagttcccc agcgacctgg acctggacat gttcaacggc
     8461 agcctggagt gcgacatgga gagcatcatc agaagcgagc tgatggacgc cgacggcctg
     8521 gacttcaact tcgacagcgg atctggagag ggaaggggaa gcctcctaac ttgcggagat
     8581 gtcgaggaga atcctggccc aatggttagt aagggagagg agttgataaa agagaatatg
     8641 cacatgaaac tgtatatgga agggacagtg gacaaccatc acttcaagtg cacctctgag
     8701 ggagaaggaa agccatacga aggcacccag actatgcgaa ttaaggtcgt ggagggaggg
     8761 cctctcccct tcgccttcga tattcttgca acatctttcc tctacgggag caaaaccttc
     8821 atcaatcaca cacaaggcat acctgacttt tttaagcaga gttttccaga gggctttacc
     8881 tgggagcgtg tgactacata cgaagatggg ggtgtgctga cggctacaca ggatacttca
     8941 cttcaggacg gttgtctgat ctacaacgtc aagatcaggg gggtcaattt cacctcgaat
     9001 ggaccagtaa tgcaaaaaaa gaccctggga tgggaagctt ttactgaaac tctctatccc
     9061 gccgacggcg gcctcgaagg cagaaacgat atggctttga agctggttgg cgggtcccat
     9121 ctaattgcca atgccaaaac cacatatagg agcaaaaagc ctgcaaagaa cctgaagatg
     9181 ccgggtgtgt actatgttga ctatagatta gagcggatca aagaggcaaa taacgaaacg
     9241 tacgtggagc agcacgaggt agccgtggcg cgctactgcg atttaccctc caaactgggc
     9301 cataaactga actgtacagg aagcggagct actaacttca gcctgctgaa gcaggctggc
     9361 gacgtggaag aaaacccagg acctatggcc aagcctttgt ctcaagaaga atccaccctc
     9421 attgaaagag caacggctac aatcaacagc atccccatct ctgaagacta cagcgtcgcc
     9481 agcgcagctc tctctagcga cggccgcatc ttcactggtg tcaatgtata tcattttact
     9541 gggggacctt gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc agctggcaac
     9601 ctgacttgta tcgtcgcgat cggaaatgag aacaggggca tcttgagccc ctgcggacgg
     9661 tgccgacagg tgcttctcga tctgcatcct gggatcaaag ccatagtgaa ggacagtgat
     9721 ggacagccga cggcagttgg gattcgtgaa ttgctgccct ctggttatgt gtgggagggc
     9781 taagaattcg atatcaagct tatcggtaat caacctctgg attacaaaat ttgtgaaaga
     9841 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     9901 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     9961 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
    10021 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
    10081 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
    10141 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
    10201 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
    10261 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
    10321 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
    10381 tgggccgcct ccccgcatcg ataccgtcga cctcgagacc tagaaaaaca tggagcaatc
    10441 acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc acaagaggag
    10501 gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac ttacaaggca
    10561 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
    10621 caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta cttccctgat
    10681 tggcagaact acacaccagg gccagggatc agatatccac tgacctttgg atggtgctac
    10741 aagctagtac cagttgagca agagaaggta gaagaagcca atgaaggaga gaacacccgc
    10801 ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt attagagtgg
    10861 aggtttgaca gccgcctagc atttcatcac atggcccgag agctgcatcc ggactgtact
    10921 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
    10981 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
    11041 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
    11101 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
    11161 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
    11221 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
    11281 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
    11341 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
    11401 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
    11461 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
    11521 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
    11581 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
    11641 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
    11701 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
    11761 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
    11821 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
    11881 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
    11941 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
    12001 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
    12061 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
    12121 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
    12181 ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg
    12241 catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga
    12301 ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg
    12361 accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg
    12421 acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct
    12481 gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga
    12541 acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg
    12601 agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact
    12661 gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
    12721 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
    12781 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
    12841 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
    12901 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
    12961 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
    13021 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
    13081 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
    13141 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
    13201 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
    13261 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
    13321 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
    13381 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
    13441 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
    13501 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
    13561 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
    13621 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
    13681 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
    13741 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
    13801 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
    13861 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
    13921 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
    13981 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
    14041 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
    14101 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
    14161 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
    14221 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
    14281 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
    14341 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
    14401 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
    14461 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
    14521 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
    14581 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
    14641 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
    14701 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
    14761 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
    14821 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
    14881 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
    14941 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
    15001 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
    15061 agaaaaata
//
