LOCUS       sequence-354929-       13174 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of dCas9-VPH from the CLYBL locus for CRISPR 
            activation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13174)
  AUTHORS   Tian R, Abarientos A, Hong J, Hashemi SH, Yan R, Drager N, Leng K, 
            Nalls MA, Singleton AB, Xu K, Faghri F, Kampmann M
  TITLE     Genome-wide CRISPRi/a screens in human neurons link lysosomal 
            failure to ferroptosis.
  JOURNAL   Nat Neurosci. 2021 May 24. pii: 10.1038/s41593-021-00862-0. doi: 
            10.1038/s41593-021-00862-0.
  PUBMED    34031600
REFERENCE   2  (bases 1 to 13174)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Neurosci. 2021 May 24. pii: 10.1038/s41593-021-00862-0. doi: 
            10.1038/s41593-021-00862-0."
COMMENT     Sequence Label: pRT043-CLYBL-DDdcas9-VPH
FEATURES             Location/Qualifiers
     source          1..13174
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..26
                     /label=SA
                     /note="splice acceptor site"
     primer_bind     complement(68..85)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    74..298
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        304..683
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        685..962
                     /label=chicken beta-actin promoter
     intron          963..1979
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1903..1925
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1987..2006
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2050..2526
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="MISLIAALAVDYVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRH
                     TWESIGRPLPGRKNIILSSQPSTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVIEQFL
                     PKAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     CDS             2551..2577
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2581..2601
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2611..2634,2635..2637,2638..5193,5194..5196,
                     5197..6711)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the N863A mutation in the HNH 
                     catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             join(2611..2634,2635..2637,2638..5124,5125..5127,
                     5128..6711)
                     /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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6715..6735
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6772..6882
                     /codon_start=1
                     /product="3 tandem repeats of the minimal activation domain
                     of herpes simplex virus VP16 (Cheng et al., 2013)"
                     /label=VP48
                     /translation="DALDDFDLDMLPADALDDFDLDMLPADALDDFDLDML"
     CDS             6934..7044
                     /codon_start=1
                     /product="3 tandem repeats of the minimal activation domain
                     of herpes simplex virus VP16 (Cheng et al., 2013)"
                     /label=VP48
                     /translation="DALDDFDLDMLPADALDDFDLDMLPADALDDFDLDML"
     CDS             7096..7206
                     /codon_start=1
                     /product="3 tandem repeats of the minimal activation domain
                     of herpes simplex virus VP16 (Cheng et al., 2013)"
                     /label=VP48
                     /translation="DALDDFDLDMLPADALDDFDLDMLPADALDDFDLDML"
     CDS             7258..7368
                     /codon_start=1
                     /product="3 tandem repeats of the minimal activation domain
                     of herpes simplex virus VP16 (Cheng et al., 2013)"
                     /label=VP48
                     /translation="DALDDFDLDMLPADALDDFDLDMLPADALDDFDLDML"
     CDS             7417..7959
                     /codon_start=1
                     /product="C-terminal portion of the p65 subunit of mouse 
                     NF-kappa-B"
                     /label=p65
                     /translation="PSGQISNQALALAPSSAPVLAQTMVPSSAMVPLAQPPAPAPVLTP
                     GPPQSLSAPVPKSTQAGEGTLSEALLHLQFDADEDLGALLGNSTDPGVFTDLASVDNSE
                     FQQLLNQGVSMSHSTAEPMLMEYPEAITRLVTGSQRPPDPAPTPLGTSGLPNGLSGDED
                     FSSIADMDFSALLSQISS"
     CDS             7984..8355
                     /codon_start=1
                     /product="C-terminal activation domain from the human heat 
                     shock transcription factor HSF1"
                     /label=HSF1
                     /translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPR
                     PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAED
                     PTISLLTGSEPPKAKDPTVS"
     CDS             8377..8430
                     /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             join(8437..8439,8440..8442,8443..9156)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(8437..8439,8440..8442,8443..9153)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(8743..8762)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     9090..9111
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(9246..9265)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    9311..9366
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(9365..9384)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(10698..11286)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10778..10797)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(11457..12248,12249..12317))
                     /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     12080..12099
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ctgacctctt ctcttcctcc cacagggcct cgagagatct ggcagcggat agaaacccgc
       61 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
      121 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
      181 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
      241 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatgggt
      301 ctcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
      361 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      421 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      481 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      541 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      601 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      661 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      721 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      781 cgatgggggc gggggggggg ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc
      841 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
      901 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
      961 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
     1021 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
     1081 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
     1141 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggggtgcgt
     1201 gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc gctgcccggc ggctgtgagc
     1261 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt gcgcgagggg agcgcggccg
     1321 ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa caaaggctgc gtgcggggtg
     1381 tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt cgggctgcaa ccccccctgc
     1441 acccccctcc ccgagttgct gagcacggcc cggcttcggg tgcggggctc cgtacggggc
     1501 gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca ggtgggggtg ccgggcgggg
     1561 cggggccgcc tcgggccggg gagggctcgg gggaggggcg cggcggcccc cggagcgccg
     1621 gcggctgtcg aggcgcggcg agccgcagcc attgcctttt atggtaatcg tgcgagaggg
     1681 cgcagggact tcctttgtcc caaatctgtg cggagccgaa atctgggagg cgccgccgca
     1741 ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg caggaaggaa atgggcgggg
     1801 agggccttcg tgcgtcgccg cgccgccgtc cccttctccc tctccagcct cggggctgtc
     1861 cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc ttctggcgtg
     1921 tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc
     1981 tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa tgtcgacgct
     2041 agcgccacca tgatcagtct gattgcggcg ttagcggtag attacgttat cggcatggaa
     2101 aacgccatgc cgtggaacct gcctgccgat ctcgcctggt ttaaacgcaa caccttaaat
     2161 aaacccgtga ttatgggccg ccatacctgg gaatcaatcg gtcgtccgtt gccaggacgc
     2221 aaaaatatta tcctcagcag tcaaccgagt acggacgatc gcgtaacgtg ggtgaagtcg
     2281 gtggatgaag ccatcgcggc gtgtggtgac gtaccagaaa tcatggtgat tggcggcggt
     2341 cgcgttattg aacagttctt gccaaaagcg caaaaactgt atctgacgca tatcgacgca
     2401 gaagtggaag gcgacaccca tttcccggat tacgagccgg atgactggga atcggtattc
     2461 agcgaattcc acgatgctga tgcgcagaac tctcacagct attgctttga gattctggag
     2521 cggcgaggct ccggaaccgg tgccaccatg tacccatacg atgttccaga ttacgcttcg
     2581 ccgaagaaaa agcgcaaggt cgaagcgtcc gacaagaagt acagcatcgg cctggccatc
     2641 ggcaccaact ctgtgggctg ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa
     2701 ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga agaacctgat cggagccctg
     2761 ctgttcgaca gcggcgaaac agccgaggcc acccggctga agagaaccgc cagaagaaga
     2821 tacaccagac ggaagaaccg gatctgctat ctgcaagaga tcttcagcaa cgagatggcc
     2881 aaggtggacg acagcttctt ccacagactg gaagagtcct tcctggtgga agaggataag
     2941 aagcacgagc ggcaccccat cttcggcaac atcgtggacg aggtggccta ccacgagaag
     3001 taccccacca tctaccacct gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg
     3061 cggctgatct atctggccct ggcccacatg atcaagttcc ggggccactt cctgatcgag
     3121 ggcgacctga accccgacaa cagcgacgtg gacaagctgt tcatccagct ggtgcagacc
     3181 tacaaccagc tgttcgagga aaaccccatc aacgccagcg gcgtggacgc caaggccatc
     3241 ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc tgatcgccca gctgcccggc
     3301 gagaagaaga atggcctgtt cggcaacctg attgccctga gcctgggcct gacccccaac
     3361 ttcaagagca acttcgacct ggccgaggat gccaaactgc agctgagcaa ggacacctac
     3421 gacgacgacc tggacaacct gctggcccag atcggcgacc agtacgccga cctgtttctg
     3481 gccgccaaga acctgtccga cgccatcctg ctgagcgaca tcctgagagt gaacaccgag
     3541 atcaccaagg cccccctgag cgcctctatg atcaagagat acgacgagca ccaccaggac
     3601 ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg agaagtacaa agagattttc
     3661 ttcgaccaga gcaagaacgg ctacgccggc tacattgacg gcggagccag ccaggaagag
     3721 ttctacaagt tcatcaagcc catcctggaa aagatggacg gcaccgagga actgctcgtg
     3781 aagctgaaca gagaggacct gctgcggaag cagcggacct tcgacaacgg cagcatcccc
     3841 caccagatcc acctgggaga gctgcacgcc attctgcggc ggcaggaaga tttttaccca
     3901 ttcctgaagg acaaccggga aaagatcgag aagatcctga ccttccgcat cccctactac
     3961 gtgggccctc tggccagggg aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa
     4021 accatcaccc cctggaactt cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc
     4081 atcgagcgga tgaccaactt cgataagaac ctgcccaacg agaaggtgct gcccaagcac
     4141 agcctgctgt acgagtactt caccgtgtat aacgagctga ccaaagtgaa atacgtgacc
     4201 gagggaatga gaaagcccgc cttcctgagc ggcgagcaga aaaaggccat cgtggacctg
     4261 ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga aagaggacta cttcaagaaa
     4321 atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg
     4381 ggcacatacc acgatctgct gaaaattatc aaggacaagg acttcctgga caatgaggaa
     4441 aacgaggaca ttctggaaga tatcgtgctg accctgacac tgtttgagga cagagagatg
     4501 atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg acaaagtgat gaagcagctg
     4561 aagcggcgga gatacaccgg ctggggcagg ctgagccgga agctgatcaa cggcatccgg
     4621 gacaagcagt ccggcaagac aatcctggat ttcctgaagt ccgacggctt cgccaacaga
     4681 aacttcatgc agctgatcca cgacgacagc ctgaccttta aagaggacat ccagaaagcc
     4741 caggtgtccg gccagggcga tagcctgcac gagcacattg ccaatctggc cggcagcccc
     4801 gccattaaga agggcatcct gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg
     4861 ggccggcaca agcccgagaa catcgtgatc gaaatggcca gagagaacca gaccacccag
     4921 aagggacaga agaacagccg cgagagaatg aagcggatcg aagagggcat caaagagctg
     4981 ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc agctgcagaa cgagaagctg
     5041 tacctgtact acctgcagaa tgggcgggat atgtacgtgg accaggaact ggacatcaac
     5101 cggctgtccg actacgatgt cgacgccatc gtgcctcaga gctttctgaa ggacgactcc
     5161 atcgacaaca aggtgctgac cagaagcgac aagaaccggg gcaagagcga caacgtgccc
     5221 tccgaagagg tcgtgaagaa gatgaagaac tactggcggc agctgctgaa cgccaagctg
     5281 attacccaga gaaagttcga caatctgacc aaggccgaga gaggcggcct gagcgaactg
     5341 gataaggccg gcttcatcaa gagacagctg gtggaaaccc ggcagatcac aaagcacgtg
     5401 gcacagatcc tggactcccg gatgaacact aagtacgacg agaatgacaa gctgatccgg
     5461 gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg atttccggaa ggatttccag
     5521 ttttacaaag tgcgcgagat caacaactac caccacgccc acgacgccta cctgaacgcc
     5581 gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc
     5641 gactacaagg tgtacgacgt gcggaagatg atcgccaaga gcgagcagga aatcggcaag
     5701 gctaccgcca agtacttctt ctacagcaac atcatgaact ttttcaagac cgagattacc
     5761 ctggccaacg gcgagatccg gaagcggcct ctgatcgaga caaacggcga aaccggggag
     5821 atcgtgtggg ataagggccg ggattttgcc accgtgcgga aagtgctgag catgccccaa
     5881 gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct tcagcaaaga gtctatcctg
     5941 cccaagagga acagcgataa gctgatcgcc agaaagaagg actgggaccc taagaagtac
     6001 ggcggcttcg acagccccac cgtggcctat tctgtgctgg tggtggccaa agtggaaaag
     6061 ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg ggatcaccat catggaaaga
     6121 agcagcttcg agaagaatcc catcgacttt ctggaagcca agggctacaa agaagtgaaa
     6181 aaggacctga tcatcaagct gcctaagtac tccctgttcg agctggaaaa cggccggaag
     6241 agaatgctgg cctctgccgg cgaactgcag aagggaaacg aactggccct gccctccaaa
     6301 tatgtgaact tcctgtacct ggccagccac tatgagaagc tgaagggctc ccccgaggat
     6361 aatgagcaga aacagctgtt tgtggaacag cacaagcact acctggacga gatcatcgag
     6421 cagatcagcg agttctccaa gagagtgatc ctggccgacg ctaatctgga caaagtgctg
     6481 tccgcctaca acaagcaccg ggataagccc atcagagagc aggccgagaa tatcatccac
     6541 ctgtttaccc tgaccaatct gggagcccct gccgccttca agtactttga caccaccatc
     6601 gaccggaaga ggtacaccag caccaaagag gtgctggacg ccaccctgat ccaccagagc
     6661 atcaccggcc tgtacgagac acggatcgac ctgtctcagc tgggaggcga cagccccaag
     6721 aagaagagaa aggtggaggc cagcctcgag tgcgaaagcg gcgggccggc cgacgccctt
     6781 gacgattttg acttagacat gctcccagcc gatgcccttg acgactttga ccttgatatg
     6841 ctgcctgctg acgctcttga cgattttgac cttgacatgc tccccgggtc actacccagc
     6901 gctagcgtcg agtgcgaaag cggcgggccg gccgacgccc ttgacgattt tgacttagac
     6961 atgctcccag ccgatgccct tgacgacttt gaccttgata tgctgcctgc tgacgctctt
     7021 gacgattttg accttgacat gctccccggg tcactaccca gcgctagcgt cgagtgcgaa
     7081 agcggcgggc cggccgacgc ccttgacgat tttgacttag acatgctccc agccgatgcc
     7141 cttgacgact ttgaccttga tatgctgcct gctgacgctc ttgacgattt tgaccttgac
     7201 atgctccccg ggtcactacc cagcgctagc gtcgagtgcg aaagcggcgg gccggccgac
     7261 gcccttgacg attttgactt agacatgctc ccagccgatg cccttgacga ctttgacctt
     7321 gatatgctgc ctgctgacgc tcttgacgat tttgaccttg acatgctccc cgggtcacta
     7381 cccagcgcta gcgtcgagtt cgaaggcagc ggcgggcctt cagggcagat cagcaaccag
     7441 gccctggctc tggcccctag ctccgctcca gtgctggccc agactatggt gccctctagt
     7501 gctatggtgc ctctggccca gccacctgct ccagcccctg tgctgacccc aggaccaccc
     7561 cagtcactga gcgctccagt gcccaagtct acacaggccg gcgaggggac tctgagtgaa
     7621 gctctgctgc acctgcagtt cgacgctgat gaggacctgg gagctctgct ggggaacagc
     7681 accgatcccg gagtgttcac agatctggcc tccgtggaca actctgagtt tcagcagctg
     7741 ctgaatcagg gcgtgtccat gtctcatagt acagccgaac caatgctgat ggagtacccc
     7801 gaagccatta cccggctggt gaccggcagc cagcggcccc ccgaccccgc tccaactccc
     7861 ctgggaacca gcggcctgcc taatgggctg tccggagatg aagacttctc aagcatcgct
     7921 gatatggact ttagtgccct gctgtcacag atttcctcta gtgggcaggg aggaggtgga
     7981 agcggcttca gcgtggacac cagtgccctg ctggacctgt tcagcccctc ggtgaccgtg
     8041 cccgacatga gcctgcctga ccttgacagc agcctggcca gtatccaaga gctcctgtct
     8101 ccccaggagc cccccaggcc tcccgaggca gagaacagca gcccggattc agggaagcag
     8161 ctggtgcact acacagcgca gccgctgttc ctgctggacc ccggctccgt ggacaccggg
     8221 agcaacgacc tgccggtgct gtttgagctg ggagagggct cctacttctc cgaaggggac
     8281 ggcttcgccg aggaccccac catctccctg ctgacaggct cggagcctcc caaagccaag
     8341 gaccccactg tctcctctag cgtcgacggc tccggagagg gcagaggaag tctgctaaca
     8401 tgcggtgacg tcgaggagaa tcctggccca atcgatatgg tgagcaaggg cgaggagctg
     8461 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
     8521 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
     8581 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
     8641 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
     8701 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
     8761 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
     8821 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
     8881 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
     8941 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     9001 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     9061 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     9121 gggatcactc tcggcatgga cgagctgtac aagtaaacgc gtgaattcac tcctcaggtg
     9181 caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac
     9241 tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg
     9301 acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt
     9361 ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg
     9421 gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa
     9481 agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc
     9541 ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc
     9601 ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca
     9661 gtcatagctg tccctcttct cttatggaga tcgatatctc ctcatccttc caacataaat
     9721 atattttggg attatatcaa cattcaatgt tacttaaagt gaccttgtaa atattttcac
     9781 aactgagcca tgtttgattt gtatacttat gtttacttta ctgtttttcc tgaagttaat
     9841 aattgccttg aatttattta tttctttaaa aatgtttcat tactcaggac tgtagtttac
     9901 attacgattc tttgtgttat acagttgatg ggtttctttt ctttcttaat ttctttaaaa
     9961 aatagagatg gggtcttact atattaccca ggctggtctt gaagtcctgg gctcaagtga
    10021 tcttcctgtc tcagcctacc aagtagctga gactataggt gcaaaaaagc cactatacct
    10081 ggctagttta caggttttaa caaatgcatt atgccacgta tccattatta caggatcaca
    10141 caagatattt tcattaccct gaaagcatcc ctgtgttcca ccaattcatc ctgcctccat
    10201 gagccgctgg caaccactga tctctatagt tttgcctttt ctaaaatgtc atataattgg
    10261 aatcatacag tctgtagcat tttcagacta gcttttaaaa tttggcaata tgcatttaag
    10321 gttcctcctt aaatgtgaag ggcatagcca atgtggcttg atagctcatt tctttttatt
    10381 ggtgaatatt tcattgtctg gatgttccac agtttgttta tccattcacc tattcaattt
    10441 gctttttttc tgtgtatcta tcactaattc aatactggac tctccaacag agccgtcccg
    10501 cttccctaag gccctgctct gggcttctgg gtttgagtcc ttggcaagcc caggagaggc
    10561 gctcaggctt ccctgtcccc cttcctcgtc caccatctca tgcccctggc tctcctgccc
    10621 cttccctaca ggggttcctg gctctgctct aagggcgaat tcgccaggaa ccgtaaaaag
    10681 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
    10741 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
    10801 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
    10861 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
    10921 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
    10981 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
    11041 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
    11101 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
    11161 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
    11221 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
    11281 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
    11341 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
    11401 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
    11461 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
    11521 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
    11581 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
    11641 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
    11701 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
    11761 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
    11821 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
    11881 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
    11941 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
    12001 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
    12061 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
    12121 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
    12181 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
    12241 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
    12301 aaatgttgaa tactcatact cttccttttt caatagcagg agaatcatgc caatgggcca
    12361 atatacattc tgacccacag tttcataata aaataaaatg gttgtggttg taagccacta
    12421 tgtttcagag tggtttgtta cacagcaata aataactaat atagtaggca taccatcaag
    12481 tccaaagtag gtagagaaga atgtaaatag cagagcaaaa cagcatgact ggtggctggg
    12541 aggcttaaaa ctgggacagg atcagagtca tgaaagaagt caaagaaatg gttcagaagt
    12601 aaggctgaga ctgacttaca aaagctgaaa gtccctttaa gttggtgttt ggtgcattgg
    12661 caggggcagg tatggtgact taaaagagcc atgctcaaca agatcaagca caacacaatc
    12721 acgggtcacc ccagcagacc ttagcgagtc tagccatttc tttggtggtg gtcacagtca
    12781 tgcttcagcc cagtttccac ttggacaaat ggtacatatt ttcaatgaga tgaaaattaa
    12841 gatacaatcc atgtgctcag agagtgatca cagctctgac taaacactgt gccccaaagt
    12901 gttgaggaat tgggaaaacc tagctgagtt agtggtctct tttctgttac aataaagctc
    12961 ataatgaaaa ttagccttct ttgttcttcc ccaagtcttt ctttctagac gaaactactt
    13021 tcaactgttt taacttcctt actgttaact tccatatttc tagatagtat ggtcagactg
    13081 ttatttcttg acttttaaat gtaagatatt atctactgac ttccttctat gtaagatgag
    13141 gattgagctc tcttaccctt ctcccagcgg ccgc
//
