LOCUS       Exported               13021 bp ds-DNA     circular SYN 28-FEB-2025
DEFINITION  Dox-inducible expression of dCas9-GCN4 tail in a pibbyBac vector. 
            Genomic integration selectable with hygro.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13021)
  AUTHORS   Policarpi C, Munafo M, Tsagkris S, Carlini V, Hackett JA.
  TITLE     Systematic epigenome editing captures the context-dependent 
            instructive function of chromatin modifications.
  JOURNAL   Nat Genet 2024 Jun;56:1168-1180
  PUBMED    38724747
REFERENCE   2  (bases 1 to 13021)
  AUTHORS   Hackett Lab
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Feb 28, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..13021
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(2..105)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     728..744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1386..1764
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    1394..1412
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1430..1448
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1466..1484
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1502..1520
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1538..1556
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1574..1592
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1610..1628
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1763..1804
                     /label=OJH_364 TRE3G-dCas9
     CDS             1785..1805
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1860..5963
                     /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"
     primer_bind     1977..1996
                     /label=OJH335
     primer_bind     complement(2050..2069)
                     /label=OJH336
     primer_bind     5437..5456
                     /label=OJH770
     primer_bind     complement(5516..5535)
                     /label=OJH771
     primer_bind     5798..5819
                     /label=OJH108
     CDS             5967..5993
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     6011..6031
                     /label=OJH_365 SunTAG-3a3L_F
     CDS             6012..6032
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6039..6059
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6072..6086)
                     /label=OJH321
     primer_bind     complement(6078..6098)
                     /label=Fragment 1.REV
     CDS             6123..6179
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             6246..6302
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             6369..6425
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             6492..6548
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             6615..6671
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     polyA_signal    6838..6886
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     complement(6899..6917)
                     /label=OJH5012
     promoter        7117..7328
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             7341..7609
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             7643..8386
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             8387..8440
                     /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             8462..9481
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV
                     LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET
                     ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV
                     MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD
                     SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF
                     DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE"
     polyA_signal    9592..9713
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(11000..11016)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    11024..11040
                     /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(11048..11078)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11093..11114
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(11402..11990)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(12161..13021)
                     /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"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgc gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaaa caaaaactca
     1081 aaatttcttc tataaagtaa caaaactttt atgagggaca gccccccccc aaagccccca
     1141 gggatgtaat tacgtccctc ccccgctagg gggcagcagc gagccgcccg gggctccgct
     1201 ccggtccggc gctccccccg catccccgag ccggcagcgt gcggggacag cccgggcacg
     1261 gggaaggtgg cacgggatcg ctttcctctg aacgcttctc gctgctcttt gagcctgcag
     1321 acacctgggg ggatacgggg aaaaggcctc cacggccact agtctttcgt cttcaagaat
     1381 tcctcgagtt tactccctat cagtgataga gaacgtatga agagtttact ccctatcagt
     1441 gatagagaac gtatgcagac tttactccct atcagtgata gagaacgtat aaggagttta
     1501 ctccctatca gtgatagaga acgtatgacc agtttactcc ctatcagtga tagagaacgt
     1561 atctacagtt tactccctat cagtgataga gaacgtatat ccagtttact ccctatcagt
     1621 gatagagaac gtataagctt taggcgtgta cggtgggcgc ctataaaagc agagctcgtt
     1681 tagtgaaccg tcagatcgcc tggagcaatt ccacaacact tttgtcttat accaactttc
     1741 cgtaccactt cctaccctcg taaaggtcta gagctagcat ggcgcccaag aagaagcgca
     1801 aggtgggacg cgtctgcagg atatcaagct tgcggtaccg cgggcccggg atcgccacca
     1861 tggacaagaa gtacagcatc ggcctggcca tcggcaccaa ctctgtgggc tgggccgtga
     1921 tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc
     1981 acagcatcaa gaagaacctg atcggcgccc tgctgttcga cagcggagaa acagccgagg
     2041 ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct
     2101 atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac
     2161 tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca
     2221 acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga
     2281 aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca
     2341 tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg
     2401 tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca
     2461 tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac
     2521 ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggcaacc
     2581 tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg
     2641 atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc
     2701 agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc
     2761 tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta
     2821 tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc
     2881 agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg
     2941 gctacatcga tggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg
     3001 aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga
     3061 agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga gagctgcacg
     3121 ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg gaaaagatcg
     3181 agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca
     3241 gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag
     3301 tggtggacaa gggcgccagc gcccagagct tcatcgagcg gatgaccaac ttcgataaga
     3361 acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt
     3421 acaacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga
     3481 gcggcgagca gaaaaaagcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg
     3541 tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct
     3601 ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta
     3661 tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc
     3721 tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc
     3781 acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc ggctggggca
     3841 ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg
     3901 atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca
     3961 gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc gatagcctgc
     4021 acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc ctgcagacag
     4081 tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag aacatcgtga
     4141 tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc cgcgagagaa
     4201 tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa gaacaccccg
     4261 tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag aatgggcggg
     4321 atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat gtggacgcta
     4381 tcgtgcctca gagctttctg aaggacgact ccatcgataa caaagtgctg actcggagcg
     4441 acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga
     4501 actactggcg ccagctgctg aatgccaagc tgattaccca gaggaagttc gacaatctga
     4561 ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc aagagacagc
     4621 tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc cggatgaaca
     4681 ctaagtacga cgagaacgac aaactgatcc gggaagtgaa agtgatcacc ctgaagtcca
     4741 agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag atcaacaact
     4801 accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg atcaaaaagt
     4861 accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga
     4921 tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc ttctacagca
     4981 acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc
     5041 ctctgatcga gacaaacggc gaaacaggcg agatcgtgtg ggataagggc cgggactttg
     5101 ccaccgtgcg gaaagtgctg tctatgcccc aagtgaatat cgtgaaaaag accgaggtgc
     5161 agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgac aagctgatcg
     5221 ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc accgtggcct
     5281 attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga
     5341 aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact
     5401 ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt
     5461 actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc ggcgaactgc
     5521 agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc
     5581 actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac
     5641 agcacaaaca ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga
     5701 tcctggccga cgctaatctg gacaaggtgc tgagcgccta caacaagcac agagacaagc
     5761 ctatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat ctgggagccc
     5821 ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc agcaccaaag
     5881 aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag acacggatcg
     5941 acctgtctca gctgggaggc gacgcctatc cctatgacgt gcccgattat gccagcctgg
     6001 gcagcggctc ccccaagaaa aaacgcaagg tggaagatcc taagaaaaag cggaaagtgg
     6061 acggcattgg tagtgggagc aacggcagca gcggatccaa cggtccgact gacgccgcgg
     6121 aagaggagct tctgagcaaa aactatcacc tcgaaaacga ggttgcgcga ctgaagaaag
     6181 gaagcgggtc cggtggaagt ggctccggat ctggaggttc tggcagcgga ggtagcggca
     6241 gtggcgaaga gctccttagt aagaactatc atctggaaaa tgaggtagcg cgcttaaaga
     6301 aagggtcggg aagtggcggc agcggaagtg ggagtggagg gagcggttct ggcggttccg
     6361 gcagtggaga ggagttgctg tctaagaact accacttaga aaacgaagtc gcacggctaa
     6421 aaaaaggttc cggctccggc ggctccggtt ctggaagcgg gggctcggga tcaggtggat
     6481 ctggatcagg agaggaattg ctttccaaaa actaccacct tgagaatgag gtggccaggt
     6541 taaagaaggg gagcggctcg gggggtagtg gatcggggtc gggcgggtca ggaagcggtg
     6601 gtagcggatc tggggaggag ctgctctcga agaattacca tttggagaac gaagtggcga
     6661 gactaaagaa gggaagcggt agtggtggtt cagggtctgg ttcaggtggc agtgggtctg
     6721 ggggctcagg gtccgggacg gccggccact agccggtggc ggtggcggag gggtttaaac
     6781 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     6841 caataaaaag acagaataaa acgcacgggt gttgggtcgt ttgttcataa acgcggggtt
     6901 cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc
     6961 cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca
     7021 gccaacgtcg gggcggcagg ccctgccata gcagatctgc gaaatcggat ccagctaaat
     7081 ctaagtggat ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag
     7141 tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga aggtggcgcg
     7201 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag
     7261 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca
     7321 gaacacagct gaagcttcga ggggctcgca tctctccttc acgcgcccgc cgccctacct
     7381 gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt ggtgcctcct
     7441 gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc ctttgtccgg
     7501 cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg accctgcttg
     7561 ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc aagctgtgac
     7621 cggcgcctac tctagagccg ccatgtctag actggacaag agcaaagtca taaactctgc
     7681 tctggaatta ctcaatggag tcggtatcga aggcctgacg acaaggaaac tcgctcaaaa
     7741 gctgggagtt gagcagccta ccctgtactg gcacgtgaag aacaagcggg ccctgctcga
     7801 tgccctgcca atcgagatgc tggacaggca tcatacccac tcctgccccc tggaaggcga
     7861 gtcatggcaa gactttctgc ggaacaacgc caagtcatac cgctgtgctc tcctctcaca
     7921 tcgcgacggg gctaaagtgc atctcggcac ccgcccaaca gagaaacagt acgaaaccct
     7981 ggaaaatcag ctcgcgttcc tgtgtcagca aggcttctcc ctggagaacg cactgtacgc
     8041 tctgtccgcc gtgggccact ttacactggg ctgcgtattg gaggaacagg agcatcaagt
     8101 agcaaaagag gaaagagaga cacctaccac cgattctatg cccccacttc tgaaacaagc
     8161 aattgagctg ttcgaccggc agggagccga acctgccttc cttttcggcc tggaactaat
     8221 catatgtggc ctggagaaac agctaaagtg cgaaagcggc gggccgaccg acgcccttga
     8281 cgattttgac ttagacatgc tcccagccga tgcccttgac gactttgacc ttgatatgct
     8341 gcctgctgac gctcttgacg attttgacct tgacatgctc cccggggagg gcagaggaag
     8401 tcttctaaca tgcggtgacg tggaggagaa tcccggccct gaattcatgg atagatccgg
     8461 aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt
     8521 ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg
     8581 agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta
     8641 tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga
     8701 attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga
     8761 cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca tggatgcgat
     8821 cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg
     8881 tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg
     8941 gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat
     9001 gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa
     9061 caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt
     9121 cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat
     9181 ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct
     9241 ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa
     9301 tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg
     9361 gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt
     9421 agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata
     9481 ggccaaggaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     9541 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcagt taacttgttt
     9601 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     9661 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     9721 tggaattgac tcaaatgatg tcaattagtc tatcagaagc tatctggtct cccttccggg
     9781 ggacaagaca tccctgttta atatttaaac agcagtgttc ccaaactggg ttcttatatc
     9841 ccttgctctg gtcaaccagg ttgcagggtt tcctgtcctc acaggaacga agtccctaaa
     9901 gaaacagtgg cagccaggtt tagccccgga attgactgga ttcctttttt agggcccatt
     9961 ggtatggctt tttccccgta tccccccagg tgtctgcagg ctcaaagagc agcgagaagc
    10021 gttcagagga aagcgatccc gtgccacctt ccccgtgccc gggctgtccc cgcacgctgc
    10081 cggctcgggg atgcgggggg agcgccggac cggagcggag ccccgggcgg ctcgctgctg
    10141 ccccctagcg ggggagggac gtaattacat ccctgggggc tttggggggg ggctgtccct
    10201 gatatctata acaagaaaat atatatataa taagttatca cgtaagtaga acatgaaata
    10261 acaatataat tatcgtatga gttaaatctt aaaagtcacg taaaagataa tcatgcgtca
    10321 ttttgactca cgcggtcgtt atagttcaaa atcagtgaca cttaccgcat tgacaagcac
    10381 gcctcacggg agctccaagc ggcgactgag atgtcctaaa tgcacagcga cggattcgcg
    10441 ctatttagaa agagagagca atatttcaag aatgcatgcg tcaattttac gcagactatc
    10501 tttctagggt taatctagct gcatcaggat catatcgtcg ggtctttttt ccggctcagt
    10561 catcgcccaa gctggcgcta tctgggcatc ggggaggaag aagcccgtgc cttttcccgc
    10621 gaggttgaag cggcatggaa agagtttgcc gaggatgact gctgctgcat tgacgttgag
    10681 cgaaaacgca cgtttaccat gatgattcgg gaaggtgtgg ccatgcacgc ctttaacggt
    10741 gaactgttcg ttcaggccac ctgggatacc agttcgtcgc ggcttttccg gacacagttc
    10801 cggatggtca gcccgaagcg catcagcaac ccgaacaata ccggcgacag ccggaactgc
    10861 cgtgccggtg tgcagattaa tgacagcggt gcggcgctgg gatattacgt cagcgaggac
    10921 gggtatcctg gctggatgcc gcagaaatgg acatggatac cccgtgagtt acccggcggg
    10981 cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
    11041 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
    11101 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
    11161 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
    11221 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
    11281 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
    11341 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
    11401 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
    11461 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
    11521 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
    11581 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
    11641 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
    11701 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
    11761 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
    11821 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
    11881 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
    11941 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
    12001 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
    12061 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
    12121 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
    12181 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
    12241 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
    12301 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
    12361 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
    12421 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
    12481 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
    12541 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
    12601 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
    12661 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
    12721 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
    12781 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
    12841 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
    12901 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
    12961 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
    13021 t
//
