LOCUS       sequence-213056-       11681 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac compatible plasmid expressing dCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11681)
  AUTHORS   Yeo NC, Chavez A, Lance-Byrne A, Chan Y, Menn D, Milanova D, Kuo CC,
            Guo X, Sharma S, Tung A, Cecchi RJ, Tuttle M, Pradhan S, Lim ET, 
            Davidsohn N, Ebrahimkhani MR, Collins JJ, Lewis NE, Kiani S, Church 
            GM
  TITLE     An enhanced CRISPR repressor for targeted mammalian gene regulation.
  JOURNAL   Nat Methods. 2018 Jul 16. pii: 10.1038/s41592-018-0048-5. doi: 
            10.1038/s41592-018-0048-5.
  PUBMED    30013045
REFERENCE   2  (bases 1 to 11681)
  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 
            Methods. 2018 Jul 16. pii: 10.1038/s41592-018-0048-5. doi: 
            10.1038/s41592-018-0048-5."
COMMENT     Sequence Label: pB-CAGGS-dCas9
FEATURES             Location/Qualifiers
     source          1..11681
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(268..289)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     480..499
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     713..735
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     727..744
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     728..744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   874..936
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        1357..1736
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1738..2015
                     /label=chicken beta-actin promoter
     intron          2017..3034
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2958..2980
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3042..3061
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      3131..3140
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3137..3163,3164..3166,3167..5650,5651..5656,5657..5722,
                     5723..5725,5726..7240)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVAAIVPQSFLKDDSIDNKVLTRSDKARGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             7253..7273
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(7298..7317)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7342..7390
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        7622..7833
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      7843..7852
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             7849..8247
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    8251..8372
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8288..8307)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8342..8361
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   9135..9169
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(9660..9676)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9660..9676)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9673..9695)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9684..9700
                     /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(9708..9714,9715..9732,9733..9738))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9753..9774
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9891..9908)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10062..10650)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10142..10161)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10821..11612,11613..11681))
                     /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     11444..11463
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgt gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaac aaaaactcaa
     1081 aatttcttct ataaagtaac aaaactttta tgagggacag ccccccccca aagcccccag
     1141 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     1201 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     1261 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     1321 cacctggggg gatacgggga aaaggcctcc acggccgaca ttgattattg actagttatt
     1381 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     1441 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     1501 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     1561 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
     1621 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     1681 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtcg
     1741 aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca cccccaattt
     1801 tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg gggggggggc
     1861 gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg
     1921 cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg cggcggcggc
     1981 ggcggcccta taaaaagcga agcgcgcggc gggcggggag tcgctgcgac gctgccttcg
     2041 ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt
     2101 actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agcgcttggt
     2161 ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg
     2221 ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc
     2281 cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg cggggctttg
     2341 tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg
     2401 gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg
     2461 tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc cctccccgag ttgctgagca
     2521 cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg ccgtgccggg
     2581 cggggggtgg cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg ccggggaggg
     2641 ctcgggggag gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg cggcgagccg
     2701 cagccattgc cttttatggt aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat
     2761 ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa
     2821 gcggtgcggc gccggcagga aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg
     2881 ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg ggggacggct gccttcgggg
     2941 gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc
     3001 taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc tggttattgt
     3061 gctgtctcat cattttggca aagaatctag ataccctcgt aaaggtctag agctagcgga
     3121 tcgaaccctt gccaccatgg acaagaagta ctccattggg ctcgctatcg gcacaaacag
     3181 cgtcggctgg gccgtcatta cggacgagta caaggtgccg agcaaaaaat tcaaagttct
     3241 gggcaatacc gatcgccaca gcataaagaa gaacctcatt ggcgccctcc tgttcgactc
     3301 cggggagacg gccgaagcca cgcggctcaa aagaacagca cggcgcagat atacccgcag
     3361 aaagaatcgg atctgctacc tgcaggagat ctttagtaat gagatggcta aggtggatga
     3421 ctctttcttc cataggctgg aggagtcctt tttggtggag gaggataaaa agcacgagcg
     3481 ccacccaatc tttggcaata tcgtggacga ggtggcgtac catgaaaagt acccaaccat
     3541 atatcatctg aggaagaagc ttgtagacag tactgataag gctgacttgc ggttgatcta
     3601 tctcgcgctg gcgcatatga tcaaatttcg gggacacttc ctcatcgagg gggacctgaa
     3661 cccagacaac agcgatgtcg acaaactctt tatccaactg gttcagactt acaatcagct
     3721 tttcgaagag aacccgatca acgcatccgg agttgacgcc aaagcaatcc tgagcgctag
     3781 gctgtccaaa tcccggcggc tcgaaaacct catcgcacag ctccctgggg agaagaagaa
     3841 cggcctgttt ggtaatctta tcgccctgtc actcgggctg acccccaact ttaaatctaa
     3901 cttcgacctg gccgaagatg ccaagcttca actgagcaaa gacacctacg atgatgatct
     3961 cgacaatctg ctggcccaga tcggcgacca gtacgcagac ctttttttgg cggcaaagaa
     4021 cctgtcagac gccattctgc tgagtgatat tctgcgagtg aacacggaga tcaccaaagc
     4081 tccgctgagc gctagtatga tcaagcgcta tgatgagcac caccaagact tgactttgct
     4141 gaaggccctt gtcagacagc aactgcctga gaagtacaag gaaattttct tcgatcagtc
     4201 taaaaatggc tacgccggat acattgacgg cggagcaagc caggaggaat tttacaaatt
     4261 tattaagccc atcttggaaa aaatggacgg caccgaggag ctgctggtaa agcttaacag
     4321 agaagatctg ttgcgcaaac agcgcacttt cgacaatgga agcatccccc accagattca
     4381 cctgggcgaa ctgcacgcta tcctcaggcg gcaagaggat ttctacccct ttttgaaaga
     4441 taacagggaa aagattgaga aaatcctcac atttcggata ccctactatg taggccccct
     4501 cgcccgggga aattccagat tcgcgtggat gactcgcaaa tcagaagaga ccatcactcc
     4561 ctggaacttc gaggaagtcg tggataaggg ggcctctgcc cagtccttca tcgaaaggat
     4621 gactaacttt gataaaaatc tgcctaacga aaaggtgctt cctaaacact ctctgctgta
     4681 cgagtacttc acagtttata acgagctcac caaggtcaaa tacgtcacag aagggatgag
     4741 aaagccagca ttcctgtctg gagagcagaa gaaagctatc gtggacctcc tcttcaagac
     4801 gaaccggaaa gttaccgtga aacagctcaa agaagactat ttcaaaaaga ttgaatgttt
     4861 cgactctgtt gaaatcagcg gagtggagga tcgcttcaac gcatccctgg gaacgtatca
     4921 cgatctcctg aaaatcatta aagacaagga cttcctggac aatgaggaga acgaggacat
     4981 tcttgaggac attgtcctca cccttacgtt gtttgaagat agggagatga ttgaagaacg
     5041 cttgaaaact tacgctcatc tcttcgacga caaagtcatg aaacagctca agaggcgccg
     5101 atatacagga tgggggcggc tgtcaagaaa actgatcaat gggatccgag acaagcagag
     5161 tggaaagaca atcctggatt ttcttaagtc cgatggattt gccaaccgga acttcatgca
     5221 gttgatccat gatgactctc tcacctttaa ggaggacatc cagaaagcac aagtttctgg
     5281 ccagggggac agtcttcacg agcacatcgc taatcttgca ggtagcccag ctatcaaaaa
     5341 gggaatactg cagaccgtta aggtcgtgga tgaactcgtc aaagtaatgg gaaggcataa
     5401 gcccgagaat atcgttatcg agatggcccg agagaaccaa actacccaga agggacagaa
     5461 gaacagtagg gaaaggatga agaggattga agagggtata aaagaactgg ggtcccaaat
     5521 ccttaaggaa cacccagttg aaaacaccca gcttcagaat gagaagctct acctgtacta
     5581 cctgcagaac ggcagggaca tgtacgtgga tcaggaactg gacatcaatc ggctctccga
     5641 ctacgacgtg gctgctatcg tgccccagtc ttttctcaaa gatgattcta ttgataataa
     5701 agtgttgaca agatccgata aagctagagg gaagagtgat aacgtcccct cagaagaagt
     5761 tgtcaagaaa atgaaaaatt attggcggca gctgctgaac gccaaactga tcacacaacg
     5821 gaagttcgat aatctgacta aggctgaacg aggtggcctg tctgagttgg ataaagccgg
     5881 cttcatcaaa aggcagcttg ttgagacacg ccagatcacc aagcacgtgg cccaaattct
     5941 cgattcacgc atgaacacca agtacgatga aaatgacaaa ctgattcgag aggtgaaagt
     6001 tattactctg aagtctaagc tggtctcaga tttcagaaag gactttcagt tttataaggt
     6061 gagagagatc aacaattacc accatgcgca tgatgcctac ctgaatgcag tggtaggcac
     6121 tgcacttatc aaaaaatatc ccaagcttga atctgaattt gtttacggag actataaagt
     6181 gtacgatgtt aggaaaatga tcgcaaagtc tgagcaggaa ataggcaagg ccaccgctaa
     6241 gtacttcttt tacagcaata ttatgaattt tttcaagacc gagattacac tggccaatgg
     6301 agagattcgg aagcgaccac ttatcgaaac aaacggagaa acaggagaaa tcgtgtggga
     6361 caagggtagg gatttcgcga cagtccggaa ggtcctgtcc atgccgcagg tgaacatcgt
     6421 taaaaagacc gaagtacaga ccggaggctt ctccaaggaa agtatcctcc cgaaaaggaa
     6481 cagcgacaag ctgatcgcac gcaaaaaaga ttgggacccc aagaaatacg gcggattcga
     6541 ttctcctaca gtcgcttaca gtgtactggt tgtggccaaa gtggagaaag ggaagtctaa
     6601 aaaactcaaa agcgtcaagg aactgctggg catcacaatc atggagcgat caagcttcga
     6661 aaaaaacccc atcgactttc tcgaggcgaa aggatataaa gaggtcaaaa aagacctcat
     6721 cattaagctt cccaagtact ctctctttga gcttgaaaac ggccggaaac gaatgctcgc
     6781 tagtgcgggc gagctgcaga aaggtaacga gctggcactg ccctctaaat acgttaattt
     6841 cttgtatctg gccagccact atgaaaagct caaagggtct cccgaagata atgagcagaa
     6901 gcagctgttc gtggaacaac acaaacacta ccttgatgag atcatcgagc aaataagcga
     6961 attctccaaa agagtgatcc tcgccgacgc taacctcgat aaggtgcttt ctgcttacaa
     7021 taagcacagg gataagccca tcagggagca ggcagaaaac attatccact tgtttactct
     7081 gaccaacttg ggcgcgcctg cagccttcaa gtacttcgac accaccatag acagaaagcg
     7141 gtacacctct acaaaggagg tcctggacgc cacactgatt catcagtcaa ttacggggct
     7201 ctatgaaaca agaatcgacc tctctcagct cggtggagac agcagggctg accccaagaa
     7261 gaagaggaag gtgtgagttt aaacggggga ggctaactga aacacggaag gagacaatac
     7321 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac gggtgttggg
     7381 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     7441 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     7501 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcagat
     7561 ctgcgaaatc ggatccgcgg ccgcactgat tctagctgca gaggctccgg tgcccgtcag
     7621 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
     7681 accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
     7741 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
     7801 ctttttcgca acgggtttgc cgccagaaca caggatcctc gagccaccat ggccaagcct
     7861 ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc
     7921 atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact
     7981 ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact
     8041 gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg
     8101 ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca tcctgggatc
     8161 aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg
     8221 ccctctggtt atgtgtggga gggctaagtt aacttgttta ttgcagctta taatggttac
     8281 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     8341 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggaattgact caaatgatgt
     8401 caattagtct atcagaagct catctggtct cccttccggg ggacaagaca tccctgttta
     8461 atatttaaac agcagtgttc ccaaactggg ttcttatatc ccttgctctg gtcaaccagg
     8521 ttgcagggtt tcctgtcctc acaggaacga agtccctaaa gaaacagtgg cagccaggtt
     8581 tagccccgga attgactgga ttcctttttt agggcccatt ggtatggctt tttccccgta
     8641 tccccccagg tgtctgcagg ctcaaagagc agcgagaagc gttcagagga aagcgatccc
     8701 gtgccacctt ccccgtgccc gggctgtccc cgcacgctgc cggctcgggg atgcgggggg
     8761 agcgccggac cggagcggag ccccgggcgg ctcgctgctg ccccctagcg ggggagggac
     8821 gtaattacat ccctgggggc tttggggggg ggctgtccct gatatctata acaagaaaat
     8881 atatatataa taagttatca cgtaagtaga acatgaaata acaatataat tatcgtatga
     8941 gttaaatctt aaaagtcacg taaaagataa tcatgcgtca ttttgactca cgcggtcgtt
     9001 atagttcaaa atcagtgaca cttaccgcat tgacaagcac gcctcacggg agctccaagc
     9061 ggcgactgag atgtcctaaa tgcacagcga cggattcgcg ctatttagaa agagagagca
     9121 atatttcaag aatgcatgcg tcaattttac gcagactatc tttctagggt taatctagct
     9181 gcatcaggat catatcgtcg ggtctttttt ccggctcagt catcgcccaa gctggcgcta
     9241 tctgggcatc ggggaggaag aagcccgtgc cttttcccgc gaggttgaag cggcatggaa
     9301 agagtttgcc gaggatgact gctgctgcat tgacgttgag cgaaaacgca cgtttaccat
     9361 gatgattcgg gaaggtgtgg ccatgcacgc ctttaacggt gaactgttcg ttcaggccac
     9421 ctgggatacc agttcgtcgc ggcttttccg gacacagttc cggatggtca gcccgaagcg
     9481 catcagcaac ccgaacaata ccggcgacag ccggaactgc cgtgccggtg tgcagattaa
     9541 tgacagcggt gcggcgctgg gatattacgt cagcgaggac gggtatcctg gctggatgcc
     9601 gcagaaatgg acatggatac cccgtgagtt acccggcggg cgcgcttggc gtaatcatgg
     9661 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     9721 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     9781 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     9841 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     9901 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     9961 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
    10021 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
    10081 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
    10141 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
    10201 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
    10261 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
    10321 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
    10381 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
    10441 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
    10501 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
    10561 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
    10621 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
    10681 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
    10741 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
    10801 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
    10861 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
    10921 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
    10981 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
    11041 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
    11101 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
    11161 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
    11221 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
    11281 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
    11341 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
    11401 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
    11461 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
    11521 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
    11581 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
    11641 aaaaagggaa taagggcgac acggaaatgt tgaatactca t
//
