LOCUS       sequence-329788-       13160 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses dCas9 repressor KRAB-dCas9 and blasticidin resistance
             .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13160)
  AUTHORS   Morris JA, Caragine C, Daniloski Z, Domingo J, Barry T, Lu L, Davis 
            K, Ziosi M, Glinos DA, Hao S, Mimitou EP, Smibert P, Roeder K, 
            Katsevich E, Lappalainen T, Sanjana NE
  TITLE     Discovery of target genes and pathways at GWAS loci by pooled 
            single-cell CRISPR screens.
  JOURNAL   Science. 2023 May 19;380(6646):eadh7699. doi: 
            10.1126/science.adh7699. Epub 2023 May 19.
  PUBMED    37141313
REFERENCE   2  (bases 1 to 13160)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science."; 
            date: "2023-05-19"; volume: "380(6646):eadh7699. doi"; pages: " 
            10.1126/science.adh7699. Epub 2023 May 19"
COMMENT     Sequence Label: lentiCRISPRi(v1)-Blast
FEATURES             Location/Qualifiers
     source          1..13160
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2098..2142
                     /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    2440..2557
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2651..2862
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2881..2890
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2917..3102
                     /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(3124..3150,3151..3153,3154..5640,5641..5643,
                     5644..7227)
                     /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             3124..7227
                     /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             7231..7257
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             7237..7263
                     /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             7276..7296
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7303..7323
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             7372..7428
                     /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             7429..7827
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    7852..8440
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7905..7925)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8323..8334)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(8443..8459)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8444..8460)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             8965..9145
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(9171..9188)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9177..9401
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9447..9875
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9534..9553)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9744..9765
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9884..9904)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        9889..10218
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      10069..10204
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     10131..10150
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        10266..10313
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             10332..10706
                     /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    10836..10957
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10873..10892)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10927..10946
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(11006..11022)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11006..11022)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11019..11041)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11030..11046
                     /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(11054..11060,11061..11078,11079..11084))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11099..11120
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11237..11254)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11408..11996)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11488..11507)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12167..12958,12959..13027))
                     /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     12790..12809
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13028..13132)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
     1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
     1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
     1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
     1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
     1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
     1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
     1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
     1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
     2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
     2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
     2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
     2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
     2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
     2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
     2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
     2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
     2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
     2581 agatccagtt tggttaatta agctagctag gtcttgaaag gagtgggaat tggctccggt
     2641 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
     2701 ggcaattgat ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
     2761 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
     2821 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggaccggtt ctagagcgct
     2881 gccaccatgg atgctaagtc actaactgcc tggtcccgga cactggtgac cttcaaggat
     2941 gtatttgtgg acttcaccag ggaggagtgg aagctgctgg acactgctca gcagatcgtg
     3001 tacagaaatg tgatgctgga gaactataag aacctggttt ccttgggtta tcagcttact
     3061 aagccagatg tgatcctccg gttggagaag ggagaagagc ccggcggttc cggcggaggg
     3121 tcgatggaca agaagtacag catcggcctg gccatcggca ccaactctgt gggctgggcc
     3181 gtgatcaccg acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac
     3241 cggcacagca tcaagaagaa cctgatcggc gccctgctgt tcgacagcgg agaaacagcc
     3301 gaggccaccc ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc
     3361 tgctatctgc aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac
     3421 agactggaag agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc
     3481 ggcaacatcg tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga
     3541 aagaaactgg tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc
     3601 cacatgatca agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc
     3661 gacgtggaca agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac
     3721 cccatcaacg ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc
     3781 agacggctgg aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcggc
     3841 aacctgattg ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc
     3901 gaggatgcca aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg
     3961 gcccagatcg gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc
     4021 atcctgctga gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc
     4081 tctatgatca agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg
     4141 cggcagcagc tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac
     4201 gccggctaca tcgatggcgg agccagccag gaagagttct acaagttcat caagcccatc
     4261 ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg
     4321 cggaagcagc ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg
     4381 cacgccattc tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag
     4441 atcgagaaga tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac
     4501 agcagattcg cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag
     4561 gaagtggtgg acaagggcgc cagcgcccag agcttcatcg agcggatgac caacttcgat
     4621 aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc
     4681 gtgtacaacg agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc
     4741 ctgagcggcg agcagaaaaa agccatcgtg gacctgctgt tcaagaccaa ccggaaagtg
     4801 accgtgaagc agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa
     4861 atctccggcg tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa
     4921 attatcaagg acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc
     4981 gtgctgaccc tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat
     5041 gcccacctgt tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg
     5101 ggcaggctga gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc
     5161 ctggatttcc tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac
     5221 gacagcctga cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc
     5281 ctgcacgagc acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag
     5341 acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc
     5401 gtgatcgaaa tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag
     5461 agaatgaagc ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac
     5521 cccgtggaaa acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg
     5581 cgggatatgt acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac
     5641 gctatcgtgc ctcagagctt tctgaaggac gactccatcg ataacaaagt gctgactcgg
     5701 agcgacaaga accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg
     5761 aagaactact ggcgccagct gctgaatgcc aagctgatta cccagaggaa gttcgacaat
     5821 ctgaccaagg ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga
     5881 cagctggtgg aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg
     5941 aacactaagt acgacgagaa cgacaaactg atccgggaag tgaaagtgat caccctgaag
     6001 tccaagctgg tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac
     6061 aactaccacc acgcccacga cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa
     6121 aagtacccta agctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg
     6181 aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac
     6241 agcaacatca tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag
     6301 cggcctctga tcgagacaaa cggcgaaaca ggcgagatcg tgtgggataa gggccgggac
     6361 tttgccaccg tgcggaaagt gctgtctatg ccccaagtga atatcgtgaa aaagaccgag
     6421 gtgcagacag gcggcttcag caaagagtct atcctgccca agaggaacag cgacaagctg
     6481 atcgccagaa agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg
     6541 gcctattctg tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt
     6601 gtgaaagagc tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc
     6661 gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct
     6721 aagtactccc tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa
     6781 ctgcagaagg gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc
     6841 agccactatg agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg
     6901 gaacagcaca aacactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga
     6961 gtgatcctgg ccgacgctaa tctggacaag gtgctgagcg cctacaacaa gcacagagac
     7021 aagcctatca gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga
     7081 gcccctgccg ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc
     7141 aaagaggtgc tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg
     7201 atcgacctgt ctcagctggg aggcgacgcc tatccctatg acgtgcccga ttatgccagc
     7261 ctgggcagcg gctcccccaa gaaaaaacgc aaggtggaag atcctaagaa aaagcggaaa
     7321 gtggacggca ttggtagtgg gagcaacggc agcagcggat ccggatccgg cgcaacaaac
     7381 ttctctctgc tgaaacaagc cggagatgtc gaagagaatc ctggaccgat ggccaagcct
     7441 ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc
     7501 atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact
     7561 ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact
     7621 gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg
     7681 ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca tcctgggatc
     7741 aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg
     7801 ccctctggtt atgtgtggga gggctaagaa ttcgatatca agcttatcgg taatcaacct
     7861 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     7921 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     7981 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     8041 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     8101 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     8161 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     8221 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt
     8281 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     8341 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     8401 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgacctcga
     8461 gacctagaaa aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt
     8521 gcctggctag aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct
     8581 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg
     8641 ggactggaag ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac
     8701 cacacacaag gctacttccc tgattggcag aactacacac cagggccagg gatcagatat
     8761 ccactgacct ttggatggtg ctacaagcta gtaccagttg agcaagagaa ggtagaagaa
     8821 gccaatgaag gagagaacac ccgcttgtta caccctgtga gcctgcatgg gatggatgac
     8881 ccggagagag aagtattaga gtggaggttt gacagccgcc tagcatttca tcacatggcc
     8941 cgagagctgc atccggactg tactgggtct ctctggttag accagatctg agcctgggag
     9001 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     9061 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     9121 tagtcagtgt ggaaaatctc tagcagggcc cgtttaaacc cgctgatcag cctcgactgt
     9181 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     9241 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     9301 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     9361 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     9421 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     9481 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     9541 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     9601 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     9661 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     9721 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     9781 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     9841 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     9901 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     9961 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
    10021 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
    10081 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
    10141 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
    10201 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcag
    10261 cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac gacaaggtga
    10321 ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg cgcgacgtcg
    10381 ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc gtggaggacg
    10441 acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg
    10501 tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg
    10561 agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga
    10621 tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc
    10681 acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc accgccgcct
    10741 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
    10801 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
    10861 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
    10921 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
    10981 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
    11041 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
    11101 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
    11161 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
    11221 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
    11281 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
    11341 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
    11401 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
    11461 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
    11521 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
    11581 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
    11641 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
    11701 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
    11761 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
    11821 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
    11881 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
    11941 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
    12001 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
    12061 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
    12121 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
    12181 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
    12241 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
    12301 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
    12361 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
    12421 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
    12481 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
    12541 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
    12601 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
    12661 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
    12721 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
    12781 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
    12841 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
    12901 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
    12961 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
    13021 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
    13081 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
    13141 cccgaaaagt gccacctgac
//
