LOCUS       sequence-354053-       11994 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 11994)
  AUTHORS   Nghi Dang , Alissa Lance-Byrne, Angela Tung, Xiaoge Guo, Ryan J 
            Cecchi, Joanna Buchthal, Alejandro Chavez, Nan Cher Yeo
  TITLE     Generation and application of a versatile CRISPR toolkit for 
            mammalian cell engineering
  JOURNAL   Synthetic Biology, Volume 6, Issue 1, 2021
REFERENCE   2  (bases 1 to 11994)
  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: "Synthetic 
            Biology, Volume 6, Issue 1, 2021"
COMMENT     Sequence Label: NAX_CAGGS_CAS9_m4_EYFP
FEATURES             Location/Qualifiers
     source          1..11994
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          148..1164
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1088..1110
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1172..1191
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1261..1270
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1267..1293,1294..1296,1297..3780,3781..3786,3787..3852,
                     3853..3855,3856..5370)
                     /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             5383..5403
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(5428..5447)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5472..5520
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        5752..5963
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      5973..5982
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5979..5981,5982..5984,5985..6698)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6024..6045)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6285..6304)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6632..6653
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    6702..6823
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6739..6758)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6793..6812
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   7586..7620
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(8111..8127)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8111..8127)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8124..8146)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8135..8151
                     /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(8159..8165,8166..8183,8184..8189))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8204..8225
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8342..8359)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8513..9101)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8593..8612)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9272..10063,10064..10132))
                     /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     9895..9914
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10133..10237)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(10263..10718)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10400..10421)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10612..10631
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10845..10867
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     10859..10876
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     10860..10876
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   11006..11068
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        11489..11868
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc
       61 aatcagagcg gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc
      121 tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc
      181 cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag
      241 gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg
      301 cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc
      361 ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc
      421 tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg cggggctttt tgcgctccgc
      481 agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag
      541 gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg
      601 tcggtcgggc tgcaaccccc cctgcacccc cctccccgag ttgctgagca cggcccggct
      661 tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg
      721 cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg ccggggaggg ctcgggggag
      781 gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg cggcgagccg cagccattgc
      841 cttttatggt aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag
      901 ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc
      961 gccggcagga aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt
     1021 ctccctctcc agcctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc
     1081 agggcggggt tcggcttctg gcgtgtgacc ggcggctcta gagcctctgc taaccatgtt
     1141 catgccttct tctttttcct acagctcctg ggcaacgtgc tggttattgt gctgtctcat
     1201 cattttggca aagaatctag ataccctcgt aaaggtctag agctagcgga tcgaaccctt
     1261 gccaccatgg acaagaagta ctccattggg ctcgctatcg gcacaaacag cgtcggctgg
     1321 gccgtcatta cggacgagta caaggtgccg agcaaaaaat tcaaagttct gggcaatacc
     1381 gatcgccaca gcataaagaa gaacctcatt ggcgccctcc tgttcgactc cggggagacg
     1441 gccgaagcca cgcggctcaa aagaacagca cggcgcagat atacccgcag aaagaatcgg
     1501 atctgctacc tgcaggagat ctttagtaat gagatggcta aggtggatga ctctttcttc
     1561 cataggctgg aggagtcctt tttggtggag gaggataaaa agcacgagcg ccacccaatc
     1621 tttggcaata tcgtggacga ggtggcgtac catgaaaagt acccaaccat atatcatctg
     1681 aggaagaagc ttgtagacag tactgataag gctgacttgc ggttgatcta tctcgcgctg
     1741 gcgcatatga tcaaatttcg gggacacttc ctcatcgagg gggacctgaa cccagacaac
     1801 agcgatgtcg acaaactctt tatccaactg gttcagactt acaatcagct tttcgaagag
     1861 aacccgatca acgcatccgg agttgacgcc aaagcaatcc tgagcgctag gctgtccaaa
     1921 tcccggcggc tcgaaaacct catcgcacag ctccctgggg agaagaagaa cggcctgttt
     1981 ggtaatctta tcgccctgtc actcgggctg acccccaact ttaaatctaa cttcgacctg
     2041 gccgaagatg ccaagcttca actgagcaaa gacacctacg atgatgatct cgacaatctg
     2101 ctggcccaga tcggcgacca gtacgcagac ctttttttgg cggcaaagaa cctgtcagac
     2161 gccattctgc tgagtgatat tctgcgagtg aacacggaga tcaccaaagc tccgctgagc
     2221 gctagtatga tcaagcgcta tgatgagcac caccaagact tgactttgct gaaggccctt
     2281 gtcagacagc aactgcctga gaagtacaag gaaattttct tcgatcagtc taaaaatggc
     2341 tacgccggat acattgacgg cggagcaagc caggaggaat tttacaaatt tattaagccc
     2401 atcttggaaa aaatggacgg caccgaggag ctgctggtaa agcttaacag agaagatctg
     2461 ttgcgcaaac agcgcacttt cgacaatgga agcatccccc accagattca cctgggcgaa
     2521 ctgcacgcta tcctcaggcg gcaagaggat ttctacccct ttttgaaaga taacagggaa
     2581 aagattgaga aaatcctcac atttcggata ccctactatg taggccccct cgcccgggga
     2641 aattccagat tcgcgtggat gactcgcaaa tcagaagaga ccatcactcc ctggaacttc
     2701 gaggaagtcg tggataaggg ggcctctgcc cagtccttca tcgaaaggat gactaacttt
     2761 gataaaaatc tgcctaacga aaaggtgctt cctaaacact ctctgctgta cgagtacttc
     2821 acagtttata acgagctcac caaggtcaaa tacgtcacag aagggatgag aaagccagca
     2881 ttcctgtctg gagagcagaa gaaagctatc gtggacctcc tcttcaagac gaaccggaaa
     2941 gttaccgtga aacagctcaa agaagactat ttcaaaaaga ttgaatgttt cgactctgtt
     3001 gaaatcagcg gagtggagga tcgcttcaac gcatccctgg gaacgtatca cgatctcctg
     3061 aaaatcatta aagacaagga cttcctggac aatgaggaga acgaggacat tcttgaggac
     3121 attgtcctca cccttacgtt gtttgaagat agggagatga ttgaagaacg cttgaaaact
     3181 tacgctcatc tcttcgacga caaagtcatg aaacagctca agaggcgccg atatacagga
     3241 tgggggcggc tgtcaagaaa actgatcaat gggatccgag acaagcagag tggaaagaca
     3301 atcctggatt ttcttaagtc cgatggattt gccaaccgga acttcatgca gttgatccat
     3361 gatgactctc tcacctttaa ggaggacatc cagaaagcac aagtttctgg ccagggggac
     3421 agtcttcacg agcacatcgc taatcttgca ggtagcccag ctatcaaaaa gggaatactg
     3481 cagaccgtta aggtcgtgga tgaactcgtc aaagtaatgg gaaggcataa gcccgagaat
     3541 atcgttatcg agatggcccg agagaaccaa actacccaga agggacagaa gaacagtagg
     3601 gaaaggatga agaggattga agagggtata aaagaactgg ggtcccaaat ccttaaggaa
     3661 cacccagttg aaaacaccca gcttcagaat gagaagctct acctgtacta cctgcagaac
     3721 ggcagggaca tgtacgtgga tcaggaactg gacatcaatc ggctctccga ctacgacgtg
     3781 gctgctatcg tgccccagtc ttttctcaaa gatgattcta ttgataataa agtgttgaca
     3841 agatccgata aagctagagg gaagagtgat aacgtcccct cagaagaagt tgtcaagaaa
     3901 atgaaaaatt attggcggca gctgctgaac gccaaactga tcacacaacg gaagttcgat
     3961 aatctgacta aggctgaacg aggtggcctg tctgagttgg ataaagccgg cttcatcaaa
     4021 aggcagcttg ttgagacacg ccagatcacc aagcacgtgg cccaaattct cgattcacgc
     4081 atgaacacca agtacgatga aaatgacaaa ctgattcgag aggtgaaagt tattactctg
     4141 aagtctaagc tggtctcaga tttcagaaag gactttcagt tttataaggt gagagagatc
     4201 aacaattacc accatgcgca tgatgcctac ctgaatgcag tggtaggcac tgcacttatc
     4261 aaaaaatatc ccaagcttga atctgaattt gtttacggag actataaagt gtacgatgtt
     4321 aggaaaatga tcgcaaagtc tgagcaggaa ataggcaagg ccaccgctaa gtacttcttt
     4381 tacagcaata ttatgaattt tttcaagacc gagattacac tggccaatgg agagattcgg
     4441 aagcgaccac ttatcgaaac aaacggagaa acaggagaaa tcgtgtggga caagggtagg
     4501 gatttcgcga cagtccggaa ggtcctgtcc atgccgcagg tgaacatcgt taaaaagacc
     4561 gaagtacaga ccggaggctt ctccaaggaa agtatcctcc cgaaaaggaa cagcgacaag
     4621 ctgatcgcac gcaaaaaaga ttgggacccc aagaaatacg gcggattcga ttctcctaca
     4681 gtcgcttaca gtgtactggt tgtggccaaa gtggagaaag ggaagtctaa aaaactcaaa
     4741 agcgtcaagg aactgctggg catcacaatc atggagcgat caagcttcga aaaaaacccc
     4801 atcgactttc tcgaggcgaa aggatataaa gaggtcaaaa aagacctcat cattaagctt
     4861 cccaagtact ctctctttga gcttgaaaac ggccggaaac gaatgctcgc tagtgcgggc
     4921 gagctgcaga aaggtaacga gctggcactg ccctctaaat acgttaattt cttgtatctg
     4981 gccagccact atgaaaagct caaagggtct cccgaagata atgagcagaa gcagctgttc
     5041 gtggaacaac acaaacacta ccttgatgag atcatcgagc aaataagcga attctccaaa
     5101 agagtgatcc tcgccgacgc taacctcgat aaggtgcttt ctgcttacaa taagcacagg
     5161 gataagccca tcagggagca ggcagaaaac attatccact tgtttactct gaccaacttg
     5221 ggcgcgcctg cagccttcaa gtacttcgac accaccatag acagaaagcg gtacacctct
     5281 acaaaggagg tcctggacgc cacactgatt catcagtcaa ttacggggct ctatgaaaca
     5341 agaatcgacc tctctcagct cggtggagac agcagggctg accccaagaa gaagaggaag
     5401 gtgtgagttt aaacggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     5461 ccgcgctatg acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc
     5521 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     5581 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     5641 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcagat ctgcgaaatc
     5701 ggatccgcgg ccgcactgat tctagctgca gaggctccgg tgcccgtcag tgggcagagc
     5761 gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct
     5821 agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc
     5881 ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca
     5941 acgggtttgc cgccagaaca caggatcctc gagccaccat ggtgagcaag ggcgaggagc
     6001 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     6061 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     6121 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ttcggctacg
     6181 gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     6241 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     6301 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     6361 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     6421 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     6481 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     6541 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccaagc
     6601 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     6661 ccgggatcac tctcggcatg gacgagctgt acaagtaagt taacttgttt attgcagctt
     6721 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     6781 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggaattgac
     6841 tcaaatgatg tcaattagtc tatcagaagc tcatctggtc tcccttccgg gggacaagac
     6901 atccctgttt aatatttaaa cagcagtgtt cccaaactgg gttcttatat cccttgctct
     6961 ggtcaaccag gttgcagggt ttcctgtcct cacaggaacg aagtccctaa agaaacagtg
     7021 gcagccaggt ttagccccgg aattgactgg attccttttt tagggcccat tggtatggct
     7081 ttttccccgt atccccccag gtgtctgcag gctcaaagag cagcgagaag cgttcagagg
     7141 aaagcgatcc cgtgccacct tccccgtgcc cgggctgtcc ccgcacgctg ccggctcggg
     7201 gatgcggggg gagcgccgga ccggagcgga gccccgggcg gctcgctgct gccccctagc
     7261 gggggaggga cgtaattaca tccctggggg ctttgggggg gggctgtccc tgatatctat
     7321 aacaagaaaa tatatatata ataagttatc acgtaagtag aacatgaaat aacaatataa
     7381 ttatcgtatg agttaaatct taaaagtcac gtaaaagata atcatgcgtc attttgactc
     7441 acgcggtcgt tatagttcaa aatcagtgac acttaccgca ttgacaagca cgcctcacgg
     7501 gagctccaag cggcgactga gatgtcctaa atgcacagcg acggattcgc gctatttaga
     7561 aagagagagc aatatttcaa gaatgcatgc gtcaatttta cgcagactat ctttctaggg
     7621 ttaatctagc tgcatcagga tcatatcgtc gggtcttttt tccggctcag tcatcgccca
     7681 agctggcgct atctgggcat cggggaggaa gaagcccgtg ccttttcccg cgaggttgaa
     7741 gcggcatgga aagagtttgc cgaggatgac tgctgctgca ttgacgttga gcgaaaacgc
     7801 acgtttacca tgatgattcg ggaaggtgtg gccatgcacg cctttaacgg tgaactgttc
     7861 gttcaggcca cctgggatac cagttcgtcg cggcttttcc ggacacagtt ccggatggtc
     7921 agcccgaagc gcatcagcaa cccgaacaat accggcgaca gccggaactg ccgtgccggt
     7981 gtgcagatta atgacagcgg tgcggcgctg ggatattacg tcagcgagga cgggtatcct
     8041 ggctggatgc cgcagaaatg gacatggata ccccgtgagt tacccggcgg gcgcgcttgg
     8101 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     8161 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     8221 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     8281 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     8341 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     8401 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     8461 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     8521 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     8581 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     8641 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     8701 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     8761 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     8821 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     8881 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     8941 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     9001 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     9061 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     9121 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     9181 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     9241 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     9301 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     9361 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     9421 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     9481 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     9541 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     9601 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     9661 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     9721 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     9781 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     9841 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     9901 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     9961 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
    10021 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
    10081 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
    10141 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
    10201 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
    10261 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
    10321 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
    10381 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
    10441 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
    10501 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
    10561 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
    10621 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
    10681 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
    10741 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
    10801 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
    10861 taaaacgacg gccagtgagc gcgcctcgtt cattcacgtt tttgaacccg tggaggacgg
    10921 gcagactcgc ggtgcaaatg tgttttacag cgtgatggag cagatgaaga tgctcgacac
    10981 gctgcagaac acgcagctag attaacccta gaaagataat catattgtga cgtacgttaa
    11041 agataatcat gtgtaaaatt gacgcatgtg ttttatcggt ctgtatatcg aggtttattt
    11101 attaatttga atagatatta agttttatta tatttacact tacatactaa taataaattc
    11161 aacaaacaat ttatttatgt ttatttattt attaaaaaaa acaaaaactc aaaatttctt
    11221 ctataaagta acaaaacttt tatgagggac agcccccccc caaagccccc agggatgtaa
    11281 ttacgtccct cccccgctag ggggcagcag cgagccgccc ggggctccgc tccggtccgg
    11341 cgctcccccc gcatccccga gccggcagcg tgcggggaca gcccgggcac ggggaaggtg
    11401 gcacgggatc gctttcctct gaacgcttct cgctgctctt tgagcctgca gacacctggg
    11461 gggatacggg gaaaaggcct ccacggccga cattgattat tgactagtta ttaatagtaa
    11521 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
    11581 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
    11641 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
    11701 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
    11761 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
    11821 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc
    11881 cccacgttct gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta
    11941 tttatttttt aattattttg tgcagcgatg ggggcggggg gggggggggg gggg
//
