LOCUS       sequence-365385-       16589 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  encodes S. pyogenes dCas9 with both n-terminal and c-terminal fusion
            of human CBP core (aa 1084-1701) driven by EF-1alpha promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16589)
  AUTHORS   Wang K, Escobar M, Li J, Mahata B, Goell J, Shah S, Cluck M, Hilton 
            IB
  TITLE     Systematic comparison of CRISPR-based transcriptional activators 
            uncovers gene-regulatory features of enhancer-promoter interactions.
  JOURNAL   Nucleic Acids Res. 2022 Aug 12;50(14):7842-7855. doi: 
            10.1093/nar/gkac582.
  PUBMED    35849129
REFERENCE   2  (bases 1 to 16589)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2022-08-12- 12"; volume: "50"; issue: 
            "14"; pages: "7842-7855"
COMMENT     Sequence Label: CBP core-dCas9-CBP core
FEATURES             Location/Qualifiers
     source          1..16589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(344..355)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             533..766
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(792..809)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    798..1022
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1068..1496
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1155..1174)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1365..1386
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1505..1525)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1510..1839
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1690..1825
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1752..1771
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1887..1934
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1953..2327
                     /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    2457..2578
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2494..2513)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2548..2567
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2627..2643)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2627..2643)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2640..2662)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2651..2667
                     /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(2675..2681,2682..2699,2700..2705))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2720..2741
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2858..2875)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3029..3617)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3109..3128)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3788..4579,4580..4648))
                     /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     4411..4430
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4649..4753)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4828..4847)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5019..5398
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5400..5598
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5553..5573
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5616..5796
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5843..5968
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6461..6694
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6879..6923
                     /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    7221..7338
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_RNA        7470..7545
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        7607..7818
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      9460..9469
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             9703..9723
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      9736..9745
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(9742..9768,9769..9771,9772..12258,12259..12261,
                     12262..13845)
                     /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 H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             9742..13845
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             13846..13893
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     regulatory      15517..15526
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             15760..15783
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             15793..15849
                     /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             15850..16446
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
                     TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
                     QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
                     PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     16343..16363
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    join(16462..16589,1..461)
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(16515..16535)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
ORIGIN
        1 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
       61 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
      121 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
      181 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
      241 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
      301 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
      361 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
      421 ccctcagacg agtcggatct ccctttgggc cgcctccccg cgtcgacttt aagaccaatg
      481 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg
      541 ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta
      601 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
      661 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
      721 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagggc ccgtttaaac
      781 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
      841 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
      901 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
      961 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     1021 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     1081 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     1141 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     1201 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     1261 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     1321 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     1381 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     1441 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     1501 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     1561 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     1621 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     1681 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     1741 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     1801 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     1861 tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag tatatcggca
     1921 tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg
     1981 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct
     2041 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
     2101 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
     2161 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
     2221 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
     2281 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtgctacgag
     2341 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     2401 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     2461 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     2521 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     2581 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     2641 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     2701 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     2761 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     2821 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2881 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2941 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3001 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3061 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3121 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3181 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3241 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3301 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3361 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3421 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3481 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3541 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3601 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3661 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3721 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3781 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3841 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3901 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3961 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4021 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4081 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4141 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4201 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4261 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4321 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4381 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4441 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4501 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4561 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4621 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4681 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4741 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     4801 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4861 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     4921 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
     4981 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta
     5041 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     5101 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     5161 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     5221 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     5281 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     5341 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     5401 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     5461 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     5521 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     5581 ggaggtctat ataagcagcg cgttttgcct gtactgggtc tctctggtta gaccagatct
     5641 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     5701 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     5761 tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg cgcccgaaca gggacttgaa
     5821 agcgaaaggg aaaccagagg agctctctcg acgcaggact cggcttgctg aagcgcgcac
     5881 ggcaagaggc gaggggcggc gactggtgag tacgccaaaa attttgacta gcggaggcta
     5941 gaaggagaga gatgggtgcg agagcgtcag tattaagcgg gggagaatta gatcgcgatg
     6001 ggaaaaaatt cggttaaggc cagggggaaa gaaaaaatat aaattaaaac atatagtatg
     6061 ggcaagcagg gagctagaac gattcgcagt taatcctggc ctgttagaaa catcagaagg
     6121 ctgtagacaa atactgggac agctacaacc atcccttcag acaggatcag aagaacttag
     6181 atcattatat aatacagtag caaccctcta ttgtgtgcat caaaggatag agataaaaga
     6241 caccaaggaa gctttagaca agatagagga agagcaaaac aaaagtaaga ccaccgcaca
     6301 gcaagcggcc gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg
     6361 aattatataa atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa
     6421 agagaagagt ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt
     6481 tcttgggagc agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca
     6541 gacaattatt gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc
     6601 aacagcatct gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg
     6661 ctgtggaaag atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac
     6721 tcatttgcac cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga
     6781 tttggaatca cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa
     6841 tacactcctt aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg
     6901 aattagataa atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata
     6961 taaaattatt cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac
     7021 tttctatagt gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc
     7081 caaccccgag gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca
     7141 gagacagatc cattcgatta gtgaacggat cggcactgcg tgcgccaatt ctgcagacaa
     7201 atggcagtat tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg
     7261 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
     7321 acaaaaattc aaaattttcg ggtttattac agggacagca gagatccagt ttggttaatt
     7381 aaggtaccga tatcactagt gttccattaa ctgcgctaaa aataattttt aaatcttttt
     7441 tagcttcttg ctcttttttg tacgtctctg ttttagagct agaaatagca agttaaaata
     7501 aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt tgaattcgct
     7561 agctaggtct tgaaaggagt gggaattggc tccggtgccc gtcagtgggc agagcgcaca
     7621 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgatccgg tgcctagaga
     7681 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     7741 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     7801 tttgccgcca gaacacagga ccggttctag agcgccacca tgatctttaa accagaggag
     7861 ttacgccagg ccctcatgcc aaccctagaa gcactgtatc gacaggaccc agagtcatta
     7921 cctttccggc agcctgtaga tccccagctc ctcggaattc cagactattt tgacatcgta
     7981 aagaatccca tggacctctc caccatcaag cggaagctgg acacagggca ataccaagag
     8041 ccctggcagt acgtggacga cgtctggctc atgttcaaca atgcctggct ctataatcgc
     8101 aagacatccc gagtctataa gttttgcagt aagcttgcag aggtctttga gcaggaaatt
     8161 gaccctgtca tgcagtccct tggatattgc tgtggacgca agtatgagtt ttccccacag
     8221 actttgtgct gctatgggaa gcagctgtgt accattcctc gcgatgctgc ctactacagc
     8281 tatcagaata ggtatcattt ctgtgagaag tgtttcacag agatccaggg cgagaatgtg
     8341 accctgggtg acgacccttc acagccccag acgacaattt caaaggatca gtttgaaaag
     8401 aagaaaaatg ataccttaga ccccgaacct ttcgttgatt gcaaggagtg tggccggaag
     8461 atgcatcaga tttgcgttct gcactatgac atcatttggc cttcaggttt tgtgtgcgac
     8521 aactgcttga agaaaactgg cagacctcga aaagaaaaca aattcagtgc taagaggctg
     8581 cagaccacaa gactgggaaa ccacttggaa gaccgagtga acaaattttt gcggcgccag
     8641 aatcaccctg aagccgggga ggtttttgtc cgagtggtgg ccagctcaga caagacggtg
     8701 gaggtcaagc ccgggatgaa gtcacggttt gtggattctg gggaaatgtc tgaatctttc
     8761 ccatatcgaa ccaaagctct gtttgctttt gaggaaattg acggcgtgga tgtctgcttt
     8821 tttggaatgc acgtccaaga atacggctct gattgccccc ctccaaacac gaggcgtgtg
     8881 tacatttctt atctggatag tattcatttc ttccggccac gttgcctccg cacagccgtt
     8941 taccatgaga tccttattgg atatttagag tatgtgaaga aattagggta tgtgacaggg
     9001 cacatctggg cctgtcctcc aagtgaagga gatgattaca tcttccattg ccacccacct
     9061 gatcaaaaaa tacccaagcc aaaacgactg caggagtggt acaaaaagat gctggacaag
     9121 gcgtttgcag agcggatcat ccatgactac aaggatattt tcaaacaagc aactgaagac
     9181 aggctcacca gtgccaagga actgccctat tttgaaggtg atttctggcc caatgtgtta
     9241 gaagagagca ttaaggaact agaacaagaa gaagaggaga ggaaaaagga agagagcact
     9301 gcagccagtg aaaccactga gggcagtcag ggcgacagca agaatgccaa gaagaagaac
     9361 aacaagaaaa ccaacaagaa caaaagcagc atcagccgcg ccaacaagaa gaagcccagc
     9421 atgcccaacg tgtccaatga cctgtcccag aagctgtatg ccaccatgga gaagcacaag
     9481 gaggtcttct tcgtgatcca cctgcacgct gggcctgtca tcaacaccct gccccccatc
     9541 gtcgaccccg accccctgct cagctgtgac ctcatggatg ggcgcgacgc cttcctcacc
     9601 ctcgccagag acaagcactg ggagttctcc tccttgcgcc gctccaagtg gtccacgctc
     9661 tgcatgctgg tggagctgca cacccagggc caggacggat cccccaagaa gaagaggaag
     9721 gtgggatcca gcgctgccac catggacaag aagtacagca tcggcctggc catcggcacc
     9781 aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag
     9841 gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc
     9901 gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc
     9961 agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg
    10021 gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac
    10081 gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc
    10141 accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg
    10201 atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac
    10261 ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac
    10321 cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct
    10381 gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag
    10441 aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag
    10501 agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac
    10561 gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc
    10621 aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc
    10681 aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc
    10741 ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac
    10801 cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga agagttctac
    10861 aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg
    10921 aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag
    10981 atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg
    11041 aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc
    11101 cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc
    11161 accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag
    11221 cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg
    11281 ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga
    11341 atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc
    11401 aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag
    11461 tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca
    11521 taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag
    11581 gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag
    11641 gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg
    11701 cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag
    11761 cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc
    11821 atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg
    11881 tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt
    11941 aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg
    12001 cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga
    12061 cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc
    12121 cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg
    12181 tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg
    12241 tccgactacg atgtggacgc catcgtgcct cagagctttc tgaaggacga ctccatcgac
    12301 aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa
    12361 gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc
    12421 cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag
    12481 gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag
    12541 atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg
    12601 aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac
    12661 aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg
    12721 ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac
    12781 aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc
    12841 gccaagtact tcttctacag caacatcatg aactttttca agaccgagat taccctggcc
    12901 aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg
    12961 tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat
    13021 atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag
    13081 aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc
    13141 ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag
    13201 tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc
    13261 ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac
    13321 ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg
    13381 ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc caaatatgtg
    13441 aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag
    13501 cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc
    13561 agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc
    13621 tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt
    13681 accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg
    13741 aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc
    13801 ggcctgtacg agacacggat cgacctgtct cagctgggag gcgacaagcg acctgccgcc
    13861 acaaagaagg ctggacaggc taagaagaag aaaggatcca tctttaaacc agaggagtta
    13921 cgccaggccc tcatgccaac cctagaagca ctgtatcgac aggacccaga gtcattacct
    13981 ttccggcagc ctgtagatcc ccagctcctc ggaattccag actattttga catcgtaaag
    14041 aatcccatgg acctctccac catcaagcgg aagctggaca cagggcaata ccaagagccc
    14101 tggcagtacg tggacgacgt ctggctcatg ttcaacaatg cctggctcta taatcgcaag
    14161 acatcccgag tctataagtt ttgcagtaag cttgcagagg tctttgagca ggaaattgac
    14221 cctgtcatgc agtcccttgg atattgctgt ggacgcaagt atgagttttc cccacagact
    14281 ttgtgctgct atgggaagca gctgtgtacc attcctcgcg atgctgccta ctacagctat
    14341 cagaataggt atcatttctg tgagaagtgt ttcacagaga tccagggcga gaatgtgacc
    14401 ctgggtgacg acccttcaca gccccagacg acaatttcaa aggatcagtt tgaaaagaag
    14461 aaaaatgata ccttagaccc cgaacctttc gttgattgca aggagtgtgg ccggaagatg
    14521 catcagattt gcgttctgca ctatgacatc atttggcctt caggttttgt gtgcgacaac
    14581 tgcttgaaga aaactggcag acctcgaaaa gaaaacaaat tcagtgctaa gaggctgcag
    14641 accacaagac tgggaaacca cttggaagac cgagtgaaca aatttttgcg gcgccagaat
    14701 caccctgaag ccggggaggt ttttgtccga gtggtggcca gctcagacaa gacggtggag
    14761 gtcaagcccg ggatgaagtc acggtttgtg gattctgggg aaatgtctga atctttccca
    14821 tatcgaacca aagctctgtt tgcttttgag gaaattgacg gcgtggatgt ctgctttttt
    14881 ggaatgcacg tccaagaata cggctctgat tgcccccctc caaacacgag gcgtgtgtac
    14941 atttcttatc tggatagtat tcatttcttc cggccacgtt gcctccgcac agccgtttac
    15001 catgagatcc ttattggata tttagagtat gtgaagaaat tagggtatgt gacagggcac
    15061 atctgggcct gtcctccaag tgaaggagat gattacatct tccattgcca cccacctgat
    15121 caaaaaatac ccaagccaaa acgactgcag gagtggtaca aaaagatgct ggacaaggcg
    15181 tttgcagagc ggatcatcca tgactacaag gatattttca aacaagcaac tgaagacagg
    15241 ctcaccagtg ccaaggaact gccctatttt gaaggtgatt tctggcccaa tgtgttagaa
    15301 gagagcatta aggaactaga acaagaagaa gaggagagga aaaaggaaga gagcactgca
    15361 gccagtgaaa ccactgaggg cagtcagggc gacagcaaga atgccaagaa gaagaacaac
    15421 aagaaaacca acaagaacaa aagcagcatc agccgcgcca acaagaagaa gcccagcatg
    15481 cccaacgtgt ccaatgacct gtcccagaag ctgtatgcca ccatggagaa gcacaaggag
    15541 gtcttcttcg tgatccacct gcacgctggg cctgtcatca acaccctgcc ccccatcgtc
    15601 gaccccgacc ccctgctcag ctgtgacctc atggatgggc gcgacgcctt cctcaccctc
    15661 gccagagaca agcactggga gttctcctcc ttgcgccgct ccaagtggtc cacgctctgc
    15721 atgctggtgg agctgcacac ccagggccag gacggatccg attacaaaga cgatgacgat
    15781 aagggatccg gcgcaacaaa cttctctctg ctgaaacaag ccggagatgt cgaagagaat
    15841 cctggaccga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
    15901 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
    15961 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
    16021 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
    16081 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
    16141 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc
    16201 aaggagcccg cgtggttcct ggccaccgtc ggagtctcgc ccgaccacca gggcaagggt
    16261 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc
    16321 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
    16381 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
    16441 gcctgaacgc gttaagtcga caatcaacct ctggattaca aaatttgtga aagattgact
    16501 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
    16561 tatcatgcta ttgcttcccg tatggcttt
//
