LOCUS       sequence-399225-       13332 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of a CRISPRi guide targeting SF3B1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13332)
  AUTHORS   Cervia LD, Shibue T, Borah AA, Gaeta B, He L, Leung L, Li N, Moyer 
            SM, Shim BH, Dumont N, Gonzalez A, Bick NR, Kazachkova M, Dempster 
            JM, Krill-Burger JM, Piccioni F, Udeshi ND, Olive ME, Carr SA, Root 
            DE, McFarland JM, Vazquez F, Hahn WC
  TITLE     A Ubiquitination Cascade Regulating the Integrated Stress Response 
            and Survival in Carcinomas.
  JOURNAL   Cancer Discov. 2023 Mar 1;13(3):766-795. doi: 
            10.1158/2159-8290.CD-22-1230.
  PUBMED    36576405
REFERENCE   2  (bases 1 to 13332)
  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: "10.1158/2159-8290";
            journalName: "Cancer Discov"; date: "2023-03-1- 1"; volume: "13"; 
            issue: "3"; pages: "766-795"
COMMENT     Sequence Label: pXPR_023d-sgCiSF3B1
FEATURES             Location/Qualifiers
     source          1..13332
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(13..33)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        18..347
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      198..333
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     260..279
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        395..442
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             461..835
                     /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    965..1086
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1002..1021)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1056..1075
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1135..1151)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1135..1151)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1148..1170)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1159..1175
                     /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(1183..1189,1190..1207,1208..1213))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1228..1249
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1366..1383)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1537..2125)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1617..1636)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2296..3087,3088..3156))
                     /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     2919..2938
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3157..3261)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3336..3355)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3527..3906
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3908..4106
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4061..4081
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             4124..4304
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4351..4476
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4969..5202
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5387..5431
                     /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    5729..5846
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5897..6145
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     5897..5917
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     6068..6087
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        6167..6242
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        6304..6515
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     protein_bind    6541..6565
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             6595..6780
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEP"
     CDS             join(6802..6828,6829..6831,6832..9318,9319..9321,
                     9322..10905)
                     /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             6802..10905
                     /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             10909..10935
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             10915..10941
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             10954..10974
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10981..11001
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11044..11100
                     /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             11101..11697
                     /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     11594..11614
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    11713..12301
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(11766..11786)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(12184..12195)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             12373..12606
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(12632..12649)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    12638..12862
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(12908..13332,1..4)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12995..13014)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     13205..13226
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
       61 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
      121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
      181 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
      241 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
      301 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
      361 cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat cggcatagta
      421 tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagt tgaccagtgc
      481 cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct
      541 cgggttctcc cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac
      601 cctgttcatc agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg
      661 ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg
      721 ggacgcctcc gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc
      781 cctgcgcgac ccggccggca actgcgtgca cttcgtggcc gaggagcagg actgacacgt
      841 gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
      901 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
      961 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     1021 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     1081 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     1141 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     1201 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     1261 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     1321 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     1381 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     1441 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     1501 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1561 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1621 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     1681 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     1741 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     1801 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     1861 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1921 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     1981 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2041 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2101 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2161 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2221 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     2281 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     2341 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     2401 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     2461 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     2521 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     2581 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     2641 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     2701 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     2761 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     2821 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     2881 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     2941 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3001 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     3061 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     3121 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     3181 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     3241 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     3301 gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag
     3361 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     3421 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
     3481 aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg
     3541 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     3601 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     3661 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     3721 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     3781 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     3841 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     3901 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
     3961 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     4021 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
     4081 gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc tctggttaga
     4141 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     4201 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     4261 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg
     4321 gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa
     4381 gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc
     4441 ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga
     4501 tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat
     4561 atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca
     4621 tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa
     4681 gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag
     4741 ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc
     4801 accgcacagc aagcggccgc tgatcttcag acctggagga ggagatatga gggacaattg
     4861 gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac
     4921 caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt
     4981 ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt
     5041 acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat
     5101 tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag
     5161 aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc
     5221 tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct
     5281 ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac
     5341 aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga
     5401 attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct
     5461 gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt
     5521 tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac
     5581 ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga
     5641 gagagacaga gacagatcca ttcgattagt gaacggatcg gcactgcgtg cgccaattct
     5701 gcagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca
     5761 gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa
     5821 aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga gatccagttt
     5881 ggttaattaa ggtaccgagg gcctatttcc catgattcct tcatatttgc atatacgata
     5941 caaggctgtt agagagataa ttagaattaa tttgactgta aacacaaaga tattagtaca
     6001 aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt
     6061 taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata
     6121 tatcttgtgg aaaggacgaa acaccgaaga tcgccaagac tcacgagttt tagagctaga
     6181 aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt
     6241 gcttttttga attcgctagc taggtcttga aaggagtggg aattggctcc ggtgcccgtc
     6301 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt
     6361 gatccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc
     6421 tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg
     6481 ttctttttcg caacgggttt gccgccagaa cacaggaccg gttctagcat cgattgatca
     6541 acaagtttgt acaaaaaagt tggcatggat gctaagtcac taactgcctg gtcccggaca
     6601 ctggtgacct tcaaggatgt atttgtggac ttcaccaggg aggagtggaa gctgctggac
     6661 actgctcagc agatcgtgta cagaaatgtg atgctggaga actataagaa cctggtttcc
     6721 ttgggttatc agcttactaa gccagatgtg atcctccggt tggagaaggg agaagagccc
     6781 ggcggttccg gcggagggtc gatggacaag aagtacagca tcggcctggc catcggcacc
     6841 aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag
     6901 gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggcgc cctgctgttc
     6961 gacagcggag aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc
     7021 agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg
     7081 gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac
     7141 gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc
     7201 accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg
     7261 atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac
     7321 ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac
     7381 cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct
     7441 gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag
     7501 aagaatggcc tgttcggcaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag
     7561 agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac
     7621 gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc
     7681 aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc
     7741 aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc
     7801 ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac
     7861 cagagcaaga acggctacgc cggctacatc gatggcggag ccagccagga agagttctac
     7921 aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg
     7981 aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag
     8041 atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg
     8101 aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc
     8161 cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc
     8221 accccctgga acttcgagga agtggtggac aagggcgcca gcgcccagag cttcatcgag
     8281 cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg
     8341 ctgtacgagt acttcaccgt gtacaacgag ctgaccaaag tgaaatacgt gaccgaggga
     8401 atgagaaagc ccgccttcct gagcggcgag cagaaaaaag ccatcgtgga cctgctgttc
     8461 aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag
     8521 tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca
     8581 taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag
     8641 gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag
     8701 gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg
     8761 cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag
     8821 cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc
     8881 atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg
     8941 tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt
     9001 aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg
     9061 cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga
     9121 cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc
     9181 cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg
     9241 tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg
     9301 tccgactacg atgtggacgc tatcgtgcct cagagctttc tgaaggacga ctccatcgat
     9361 aacaaagtgc tgactcggag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa
     9421 gaggtcgtga agaagatgaa gaactactgg cgccagctgc tgaatgccaa gctgattacc
     9481 cagaggaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag
     9541 gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag
     9601 atcctggact cccggatgaa cactaagtac gacgagaacg acaaactgat ccgggaagtg
     9661 aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac
     9721 aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg
     9781 ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac
     9841 aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc
     9901 gccaagtact tcttctacag caacatcatg aactttttca agaccgagat taccctggcc
     9961 aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaacagg cgagatcgtg
    10021 tgggataagg gccgggactt tgccaccgtg cggaaagtgc tgtctatgcc ccaagtgaat
    10081 atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag
    10141 aggaacagcg acaagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc
    10201 ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag
    10261 tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc
    10321 ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac
    10381 ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg
    10441 ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc caaatatgtg
    10501 aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag
    10561 cagaaacagc tgtttgtgga acagcacaaa cactacctgg acgagatcat cgagcagatc
    10621 agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaggt gctgagcgcc
    10681 tacaacaagc acagagacaa gcctatcaga gagcaggccg agaatatcat ccacctgttt
    10741 accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg
    10801 aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc
    10861 ggcctgtacg agacacggat cgacctgtct cagctgggag gcgacgccta tccctatgac
    10921 gtgcccgatt atgccagcct gggcagcggc tcccccaaga aaaaacgcaa ggtggaagat
    10981 cctaagaaaa agcggaaagt ggacggcatt ggtagtggga gcaacggcag cagcggatcc
    11041 ggcgcaacaa acttctctct gctgaaacaa gccggagatg tcgaagagaa tcctggaccg
    11101 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc
    11161 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc
    11221 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
    11281 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
    11341 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
    11401 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
    11461 gcgtggttcc tggccaccgt cggagtctcg cccgaccacc agggcaaggg tctgggcagc
    11521 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
    11581 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
    11641 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaacg
    11701 cgttaagtcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
    11761 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
    11821 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
    11881 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
    11941 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
    12001 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
    12061 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
    12121 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
    12181 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
    12241 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
    12301 cgtcgacttt aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag
    12361 aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg ctttttgctt
    12421 gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc taactaggga
    12481 acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc
    12541 tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct
    12601 ctagcagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
    12661 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
    12721 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
    12781 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
    12841 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
    12901 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
    12961 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
    13021 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
    13081 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
    13141 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
    13201 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
    13261 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
    13321 aaatttaacg cg
//
