LOCUS       sequence-391224-       13061 bp    DNA     circular SYN 14-APR-2025
DEFINITION  pDEL158; transgenic construct to express cell-specific Cas9 in 
            sensory neurons; ubiquitous nrg1 EGF sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13061)
  AUTHORS   Lysko DE, Talbot WS
  TITLE     Unmyelinated sensory neurons use Neuregulin signals to promote 
            myelination of interneurons in the CNS.
  JOURNAL   Cell Rep. 2022 Nov 15;41(7):111669. doi: 
            10.1016/j.celrep.2022.111669.
  PUBMED    36384112
REFERENCE   2  (bases 1 to 13061)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Rep.";
            date: "2022-11-15"; volume: "41(7):111669. doi"; pages: " 
            10.1016/j.celrep.2022.111669"
COMMENT     Sequence Label: zCREST3:Cas9green; EGF_gRNA
FEATURES             Location/Qualifiers
     source          1..13061
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        17..121
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(122..190,191..982)
                     /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     complement(340..359)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1153..1741
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1642..1661
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1895..1912
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2029..2050
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2065..2070,2071..2088,2089..2095)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2103..2119
                     /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)."
     primer_bind     2108..2130
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2127..2143
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2164..2182
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2219..2235
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2219..2235
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             join(3690..3692,3693..3695,3696..4409)
                     /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(3690..3692,3693..3695,3696..4406)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    4711..4845
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_RNA        5230..5305
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     5318..5334
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5342..5362
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    5786..5810
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        5839..5857
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             complement(join(5894..6607,6608..6610,6611..6613))
                     /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             complement(join(5897..6607,6608..6610,6611..6613))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(6620..6673)
                     /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             complement(6689..6709)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(6725..10825)
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes MGAS15252 Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDDYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFGSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLADSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQIYNQLFEENPINASRVDAKAILSARLSKSRRL
                     ENLIAQLPGEKRNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNSEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGAYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDRGMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGHSLHEQIANLAGSPAIKKGILQTVKIVDELVKVMGH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFIKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     RBS             6804..6812
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             complement(10832..10852)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      10861..10870
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(10906..10924)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10934..10950)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(10952..10976)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(11838..11858)
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(11878..11894)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     12894..12915
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
       61 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
      121 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
      181 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
      241 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
      301 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
      361 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
      421 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
      481 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
      541 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
      601 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
      661 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
      721 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
      781 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
      841 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
      901 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
      961 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     1021 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     1081 gaccaaaatg ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     1141 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     1201 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     1261 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     1321 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     1381 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     1441 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     1501 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     1561 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     1621 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     1681 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     1741 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     1801 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     1861 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     1921 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     1981 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     2041 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     2101 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg
     2161 cgcaattaac cctcactaaa gggaacaaaa gctggagctc caccgcggtg gcggccgctc
     2221 tagaactagt ggatctgctg ggcttgctga aggtaggggg tcaagaacca gaggtgtaaa
     2281 gtacttgagt aattttactt gattactgta cttaagtatt atttttgggg atttttactt
     2341 tacttgagta caattaaaaa tcaatacttt tacttttact taattacatt tttttagaaa
     2401 aaaaagtact ttttactcct tacaatttta tttacagtca aaaagtactt attttttgga
     2461 gatcacttca ttctattttc ccttgctatt accaaaccaa ttgaattgcg ctgatgccca
     2521 gtttaattta aatgttattt attctgccta tgaaaatcgt tttcacatta tatgaaattg
     2581 gtcagacatg ttcattggtc ctttggaagt gacgtcatgt cacatctatt accacaatgc
     2641 acagcacctt gacctggaaa ttagggaaat tataacagtc aatcagtgga agaaaatgga
     2701 ggaagtatgt gattcatcag cagctgcgag cagcacagtc caaaatcagc cacaggatca
     2761 agagcacccg tggccgtatc ttcgcagatc taaagcttaa atcagttgtg ttaaataaga
     2821 gacattcaaa ataaatgtaa atgagctctc caaatcagca gacttaacat tctttaaaat
     2881 gattgattca atagtgataa aaatcaggca tagccagttg taactttaga taaattacag
     2941 aaaatgtcaa atacagagaa ccgattcttt tttatgatac atccaagcac acatttaaca
     3001 caatccaggc aaaccccgaa tttcacagtc acaagcactg tttgtacaag agctttgcct
     3061 aaggacacac agtctctata agtccaggtc gttggtttca ctcttatttt aaacatgtga
     3121 catttttcct gccatcctgt cttaggctgc tgtttgcttc attccatgtc acattaaatt
     3181 cctcagtagc accttttaca cacacagcca atcttttcca gaaaattcaa ttgctttgaa
     3241 gagataatgt gtgaacaaat ccatttagaa aaggaaaatt aagaatttgt aaaatcatct
     3301 gtaaattgtt ggcattcttc tgtatatgaa catcacatca tttacaggta aaggtctggt
     3361 cattaattat atgacaattt actggtatta ttttgtgaaa ggggctattt tcaatgcgtt
     3421 catccatcct tttcatccct caaatctctc attcacgtcc ccctccccat ctgcacactt
     3481 tatctcattt ccaccctgct ggaatctgag cacttgtgca gttatcaggg ctcctgtatt
     3541 taggaggctc tgggtgtcca tgtaggggac gaacagaaac actgcagacc tttatagaag
     3601 aacaattgat aagagtcctc atacataaag actccattag taagccagtg acccaggagc
     3661 ccagaccaac agcaaagcag acagtgacca tggtgagcaa gggcgaagag ctgttcaccg
     3721 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     3781 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     3841 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     3901 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     3961 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     4021 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     4081 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     4141 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     4201 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     4261 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     4321 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     4381 ctctcggcat ggacgagctg tacaagtaaa gcggcccctc tcgagcctct agaactatag
     4441 tgagtcgtat tacgtagatc cagacatgat aagatacatt gatgagtttg gacaaaccac
     4501 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     4561 tgtaaccatt ataagctgca ataaacaaga gatctgatct agaggatcat aatcagccat
     4621 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     4681 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     4741 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4801 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcatcat cgatgcgtct
     4861 tttgttctgg tcatcaagga ggggggaatg ttctgcgcat gcctgtgggg gagggagaag
     4921 gacacgtcac tgaaaacgtc cctgcatcac accgagacac ccaatcactc aagccgagac
     4981 cagataattt tgcatatgct ttaaagtttg aaaaatacca cggtaaaccc tcacacaaac
     5041 tctggatttg agatctttca ggttttatca gtttgcaggt ttatgtcacc atgatatagg
     5101 gtcagacttg atctaaggga gctgaataag tggtttagtc actcaccacc tcccaaaaac
     5161 atacccagaa gtccctggta tatatagctc tccctccagc tcttggttcg gatgtcttgg
     5221 ccgagggagg ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact
     5281 tgaaaaagtg gcaccgagtc ggtgcttttt tggtaccgta aaacgacggc cagtgaatta
     5341 tcaactatgt ataataaagt tgctggcata cactcacttc cactaataaa atgagaatat
     5401 tttcaatgca gaacatctgt atggagcact ttagattcac acagttcacc tgatcaatca
     5461 aatataaata tgtcaagatc aaaaacaatg tgcactcacc aagtcaatgc acagacaata
     5521 tcagtgtgtg gtgtgttatt tcacagaaac taactgcaca tctaaaaata ataaacctac
     5581 cgtgagaagg agagtacaaa gtgaacgtgg catacacaga gtgagccgat ccgagcagca
     5641 aagccatgta gcaaagacaa agaaaaacat caggcaggaa atttgggcaa cattttattt
     5701 ataggcttta tcttttcaga acatcaaatc tcaacagtct ccacaggttt tgacatgttc
     5761 agttaacggt ggctgagact taatagccac tttgtacaag aaagctgggt gctatgacca
     5821 tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctccaccg
     5881 cggtggcggc cgcttacttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca
     5941 cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc agggcggact
     6001 gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct
     6061 gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt
     6121 tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt
     6181 tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct
     6241 cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc
     6301 cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga
     6361 agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg ccgtaggtca
     6421 gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgaacttca
     6481 gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc
     6541 cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcttcgc
     6601 ccttgctcac cattctagaa gggccgggat tctcttccac gtcaccgcat gtcagcagac
     6661 tgcctctgcc ctctccgcca ccagacctca cctttctctt cttcttaggg gatcccaccg
     6721 gtgagtcgcc acccagctga gacaggtcaa tccttgtttc gtacaggccg gtaatactct
     6781 gatggatcag tgtagcgtcc agcacctcct tagtggaggt gtacctcttt ctatcgattg
     6841 tagtgtcgaa gtacttgaaa gctgcaggtg cgcccaggtt tgtcagagtg aacagatgaa
     6901 tgatattttc ggcctgctcc ctgatgggct tatctctgtg tttgttgtaa gcagacagga
     6961 ccttatccag atttgcgtcg gccaggatca ctcttttaga gaactcgcta atctgttcaa
     7021 tgatctcgtc cagataatgc ttgtgctgct ccacaaacag ctgtttctgt tcgttatcct
     7081 ctggagatcc tttcagcttc tcgtagtggc ttgccagata caggaagttc acgtacttag
     7141 aaggcagggc cagttcattg cctttctgca gctctccagc gcttgccagc atgcgtttcc
     7201 tgccgttttc cagctcaaac agactgtact tgggcagttt aatgatcagg tctttcttca
     7261 cttccttata acctttggcc tccagaaaat cgattgggtt cttctcgaaa gagcttctct
     7321 ccataattgt gatacccagc agctccttga cagatttcag tttcttgctt ttgcccttct
     7381 ccaccttagc gaccaccagg acggagtatg ccacagtggg agagtcaaag cctccatact
     7441 tcttagggtc ccagtctttc tttctagcga tcagcttatc agagttgcgt tttggcagaa
     7501 tactttcttt tgagaaaccg cctgtctgca cctcagtttt cttgacgata ttcacttgag
     7561 gcatgctcag gaccttcctc acggttgcaa aatctctgcc tttgtcccac acgatttcgc
     7621 ctgtctctcc gtttgtctca atcaggggcc ttttgcggat ttctccattg gccagggtga
     7681 tctctgtctt aaagaaattc atgatgtttg agtaaaagaa atacttggca gtagcttttc
     7741 caatttcctg ctcggacttg gcgatcattt ttctgacatc gtacacctta tagtctccgt
     7801 acacgaattc gctctccagt tttgggtatt tcttgatcag tgcggtaccg accacggcgt
     7861 tcaggtatgc gtcatgggca tggtgataat tgttaatctc gcggacctta taaaactgga
     7921 aatctttcct aaagtcgctg accagtttac tcttcagggt aatgaccttc acttctctga
     7981 tcagtttgtc attctcatcg tactttgtgt tcatgcgact atccagaatc tgtgcgacgt
     8041 gtttagtgat ctggcgtgtt tccaccagct gcctcttgat gaatccagct ttgtccagtt
     8101 ctgacagtcc accgcgctct gcttttgtca ggttatcaaa cttcctctga gtgatcagtt
     8161 tggcattcag cagctgtctc cagtagttct tcattttctt gaccacttcc tcactaggga
     8221 cattatctga ttttcctctg tttttgtcgg acctggtcag caccttgttg tcaatgctgt
     8281 catctttgat gaaactctgg gggacgatat gatccacgtc gtaatcagac agcctgttaa
     8341 tatccagctc ctggtccacg tacatatctc ttccattctg cagatagtac agatacagct
     8401 tctcattctg cagctgtgtg ttttccactg ggtgctcttt caggatctgg ctacccagct
     8461 ccttaatgcc ttcctcgatt cttttcatcc tctctctact gttcttctga cccttctggg
     8521 ttgtctgatt ttcgcgagcc atctcaatca cgatgttttc gggcttatgg cccatgactt
     8581 tcaccagctc atcgacgatt ttcactgtct gcagaatgcc tttcttgatg gctgggcttc
     8641 cagccaggtt tgcaatctgc tcatgcaggg agtggccctg tccagacacc tgagccttct
     8701 ggatgtcctc tttgaaggtc agactgtcat cgtgaatcag ctgcataaaa ttgcggttgg
     8761 cgaagccatc acttttcaga aagtccagga ttgtctttcc tgactgttta tccctaatgc
     8821 cgttgatcag cttcctgctc agtctacccc agccagtgta gcgccttctc ttcagctgtt
     8881 tcatcacctt gtcatcgaac agatgagcat aggttttcag tctttcctcg atcattcctc
     8941 tgtcctcaaa cagggtcagt gtcagcacaa tgtcttccag gatatcctca ttttcctcgt
     9001 tgtccaggaa gtctttatcc ttaatgatct tcagcagatc gtggtaagcg cccaggcttg
     9061 cgttaaatct gtcttcgact ccgctgatct ccacggaatc gaagcattca attttcttga
     9121 agtagtcctc cttcagctgc ttgacagtca ctttcctgtt ggttttaaac agcagatcca
     9181 cgattgcttt cttctgctct cctgacagga aggctggctt tctcatgccc tcggtgacgt
     9241 atttcacctt tgtcagttcg ttatagacag tgaaatactc gtacagcagg gagtgcttag
     9301 gcagcacttt ctcattgggc aggtttttgt caaaattagt cattctctcg atgaaggact
     9361 gtgcagaggc gcccttatcg accacttcct cgaagttcca aggagtaatg gtttcctcgg
     9421 atttgcgagt catccaggca aatctgctgt ttccgcgagc cagtggaccc acatagtaag
     9481 ggattctgaa ggtcagaatt ttttcgatct tctcgcggtt atctttcaga aaggggtaga
     9541 agtcttcctg gcgcctcagg atagcgtgca gctctcccag atgaatctgg tgtgggatgg
     9601 aaccgttatc aaatgtgcgc tgcttcctca gcaggtcctc tctattcagt ttcaccagca
     9661 gttcctcagt tccgtccatc ttctccagaa taggtttgat aaacttgtag aattcctcct
     9721 gtgatgcgcc tccgtcgata taaccggcgt agccgttctt ggactgatca aagaaaattt
     9781 ccttgtactt ctctggcagc tgctgcctga ccagagcctt cagcagggtc agatcctgat
     9841 ggtgctcgtc atatctcttg atcatgctgg cactcagagg tgcttttgta atctcgctgt
     9901 tcactctcag aatatctgac agcaggattg cgtcggacag atttttggca gccaggaaca
     9961 ggtcagcgta ctgatctccg atctgtgcca gcaggttatc caggtcatcg tcatatgtgt
    10021 cctttgacag ctgcagtttg gcgtcctcag ccagatcaaa attgcttttg aagttaggag
    10081 tcaggcccag actcagagcg atcagatttc caaacagacc gttcctcttt tctccgggca
    10141 gctgtgcaat caggttctcc agtctgcgtg acttggacag cctggctgac aggatagctt
    10201 ttgcgtccac tctggaagca ttaatagggt tttcctcaaa cagctgattg taaatctgga
    10261 ccagctggat gaacagcttg tccacatcgg aattgtcggg gttcagatcg ccctcgatca
    10321 gaaaatgacc cctgaactta atcatgtggg ccagagccag atagatcagt ctcaggtcgg
    10381 ctttatcagt agagtcagcc agtttcttgc gcagatgata gatggttggg tatttttcgt
    10441 gataggcgac ctcgtccaca atgtttccaa agataggatg gcgctcgtgt ttcttatctt
    10501 cctcgaccag gaaactttcc tccagcctat gaaagaatga atcgtccacc ttagccattt
    10561 cgttagagaa aatctcctgc aggtaacaga ttctattctt ccttctggta tagcgccttc
    10621 ttgctgttct tttcaggcgt gttgcttcgg cagtctctcc agaaccgaac agcagggctc
    10681 caatcagatt tttcttgatg ctgtgcctgt cggtgttacc cagcacctta aatttcttgc
    10741 tagggacctt gtagtcgtct gtaatcactg cccaacccac ggagttagtt ccaatatcca
    10801 ggccgatgct atacttctta tccatgcttc ccacctttct cttcttctta ggtggagaag
    10861 ccatggtggc gtcgacctcg agggggggcc cggtacccaa ttcgccctat agtgagtcgt
    10921 attacgcgcg ctcactggcc gtcgttttac aagcctgctt ttttgtacaa acttggcctg
    10981 ctgctggtgt catttactgg ctgaacgatg atcagcaaac tcagatgtct tatttatctt
    11041 gagctgaatc tgaaccggac tgttcagcac tgaatctata ggccgattgt gtgtgtgtgt
    11101 gtgtgtgtgt agcaacaatt ctgataaaag cttgactctc gcggcctaat tgatttattt
    11161 acagtgatcc ggtcttagtg ggaagcccct ttcatgttga ttgatcaccg caacatgcag
    11221 cagatttaac agcacaacaa acctttttgt actaatgacc caggagcatt aaagctctcg
    11281 ggcaacgtct ccattcacgc aaaagtgtgg atttgcagct cacgtgagag aggtgacttg
    11341 gtttatgtgt tcctatgaag ctgcccataa tgaacaattt ctcctaatgg caaattatcc
    11401 ctgtttctct ttcctgaagc cgaggcccct gcaaattgat gtatctgtcc tccgagggtt
    11461 tcaccgtcac attacggcgt tttaaaatat taatatgaga gtgcatttag ccaggagtag
    11521 cgaggcacgt tgctttcatg taggtcacaa gagactgact ccctctccct ctgtctgttt
    11581 ctgtctgtct ctttcatttt atctctctct ctctttcttt tctcctttct cgctctcttt
    11641 ttcatctggc tagttcagat attctgtttt tttttgtaaa taactgaatg aagagttgat
    11701 atatatattt gtgtatgtgt gtgtgtgtgt atgtttgcta ttatttagat tttctgactt
    11761 tctttgcttg gcattcagtg tgtagaggtg ccctctcaaa cactgattaa agcagacgac
    11821 gtgtcacatc ctgttaccaa cttttctata caaagttgat agcttggcgt aatcatggtc
    11881 atagctgttt cctgctcgag ggcccatctg gcctgtgttt cagacaccag ggagtctctg
    11941 ctcacgtttc ctgctatttg cagcctctct atcaagacta atacacctct tcccgcatcg
    12001 gctgcctgtg agaggctttt cagcactgca ggattgcttt tcagccccaa aagagctagg
    12061 cttgacacta acaattttga gaatcagctt ctactgaagt taaatctgag gttttacaac
    12121 tttgagtagc gtgtactggc attagattgt ctgtcttata gtttgataat taaatacaaa
    12181 cagttctaaa gcaggataaa accttgtatg catttcattt aatgtttttt gagattaaaa
    12241 gcttaaacaa gaatctctag ttttctttct tgcttttact tttacttcct taatactcaa
    12301 gtacaatttt aatggagtac ttttttactt ttactcaagt aagattctag ccagatactt
    12361 ttacttttaa ttgagtaaaa ttttccctaa gtacttgtac tttcacttga gtaaaatttt
    12421 tgagtacttt ttacacctct gtcaagaact cctggacaaa cctctgacct gtgtggaaca
    12481 gagtggatat gggtgtctga acagatattc acgtcttttg cagatcagag ggcatttctg
    12541 gtgttgtgca atgacctggg tccaacaggt aggcgtactc gtatccaaga gcattgttgg
    12601 acaggaggta gtctccattc ctataaagag gaaggaatgg caccatgtag tagccgtcat
    12661 tgtggccaat tggtgcattg gtacgtgggt agatggaccg gggaggctga tgagttctta
    12721 gccagccttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
    12781 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
    12841 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
    12901 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
    12961 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
    13021 gaattttaac aaaatattaa cgcttacaat ttaggtggca c
//
