LOCUS       sequence-354051-       11946 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 11946)
  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 11946)
  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_EBFP2
FEATURES             Location/Qualifiers
     source          1..11946
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          111..1127
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1051..1073
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1135..1154
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1224..1233
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1230..1256,1257..1259,1260..3743,3744..3749,3750..3815,
                     3816..3818,3819..5333)
                     /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             5346..5366
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(5391..5410)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5435..5483
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        5715..5926
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      5936..5945
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5942..5944,5945..5947,5948..6661)
                     /codon_start=1
                     /product="enhanced blue variant of GFP (Ai et al., 2007)"
                     /label=EBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     primer_bind     complement(5987..6008)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    6665..6786
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6702..6721)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6756..6775
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   7549..7583
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(8074..8090)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8074..8090)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8087..8109)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8098..8114
                     /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(8122..8128,8129..8146,8147..8152))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8167..8188
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8305..8322)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8476..9064)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8556..8575)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9235..10026,10027..10095))
                     /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     9858..9877
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10096..10200)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(10226..10681)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10363..10384)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10575..10594
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10808..10830
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     10822..10839
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     10823..10839
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   10969..11031
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        11452..11831
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg
       61 cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg
      121 cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct
      181 gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta
      241 attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa agccttgagg
      301 ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt
      361 gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg
      421 gcgcggggct ttttgcgctc cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc
      481 cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg
      541 ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc
      601 gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt ggcgcggggc
      661 tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg gggccgcctc
      721 gggccgggga gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag
      781 gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc
      841 ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg
      901 ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg
      961 cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg
     1021 gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct
     1081 ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg
     1141 tgctggttat tgtgctgtct catcattttg gcaaagaatc tagataccct cgtaaaggtc
     1201 tagagctagc ggatcgaacc cttgccacca tggacaagaa gtactccatt gggctcgcta
     1261 tcggcacaaa cagcgtcggc tgggccgtca ttacggacga gtacaaggtg ccgagcaaaa
     1321 aattcaaagt tctgggcaat accgatcgcc acagcataaa gaagaacctc attggcgccc
     1381 tcctgttcga ctccggggag acggccgaag ccacgcggct caaaagaaca gcacggcgca
     1441 gatatacccg cagaaagaat cggatctgct acctgcagga gatctttagt aatgagatgg
     1501 ctaaggtgga tgactctttc ttccataggc tggaggagtc ctttttggtg gaggaggata
     1561 aaaagcacga gcgccaccca atctttggca atatcgtgga cgaggtggcg taccatgaaa
     1621 agtacccaac catatatcat ctgaggaaga agcttgtaga cagtactgat aaggctgact
     1681 tgcggttgat ctatctcgcg ctggcgcata tgatcaaatt tcggggacac ttcctcatcg
     1741 agggggacct gaacccagac aacagcgatg tcgacaaact ctttatccaa ctggttcaga
     1801 cttacaatca gcttttcgaa gagaacccga tcaacgcatc cggagttgac gccaaagcaa
     1861 tcctgagcgc taggctgtcc aaatcccggc ggctcgaaaa cctcatcgca cagctccctg
     1921 gggagaagaa gaacggcctg tttggtaatc ttatcgccct gtcactcggg ctgaccccca
     1981 actttaaatc taacttcgac ctggccgaag atgccaagct tcaactgagc aaagacacct
     2041 acgatgatga tctcgacaat ctgctggccc agatcggcga ccagtacgca gacctttttt
     2101 tggcggcaaa gaacctgtca gacgccattc tgctgagtga tattctgcga gtgaacacgg
     2161 agatcaccaa agctccgctg agcgctagta tgatcaagcg ctatgatgag caccaccaag
     2221 acttgacttt gctgaaggcc cttgtcagac agcaactgcc tgagaagtac aaggaaattt
     2281 tcttcgatca gtctaaaaat ggctacgccg gatacattga cggcggagca agccaggagg
     2341 aattttacaa atttattaag cccatcttgg aaaaaatgga cggcaccgag gagctgctgg
     2401 taaagcttaa cagagaagat ctgttgcgca aacagcgcac tttcgacaat ggaagcatcc
     2461 cccaccagat tcacctgggc gaactgcacg ctatcctcag gcggcaagag gatttctacc
     2521 cctttttgaa agataacagg gaaaagattg agaaaatcct cacatttcgg ataccctact
     2581 atgtaggccc cctcgcccgg ggaaattcca gattcgcgtg gatgactcgc aaatcagaag
     2641 agaccatcac tccctggaac ttcgaggaag tcgtggataa gggggcctct gcccagtcct
     2701 tcatcgaaag gatgactaac tttgataaaa atctgcctaa cgaaaaggtg cttcctaaac
     2761 actctctgct gtacgagtac ttcacagttt ataacgagct caccaaggtc aaatacgtca
     2821 cagaagggat gagaaagcca gcattcctgt ctggagagca gaagaaagct atcgtggacc
     2881 tcctcttcaa gacgaaccgg aaagttaccg tgaaacagct caaagaagac tatttcaaaa
     2941 agattgaatg tttcgactct gttgaaatca gcggagtgga ggatcgcttc aacgcatccc
     3001 tgggaacgta tcacgatctc ctgaaaatca ttaaagacaa ggacttcctg gacaatgagg
     3061 agaacgagga cattcttgag gacattgtcc tcacccttac gttgtttgaa gatagggaga
     3121 tgattgaaga acgcttgaaa acttacgctc atctcttcga cgacaaagtc atgaaacagc
     3181 tcaagaggcg ccgatataca ggatgggggc ggctgtcaag aaaactgatc aatgggatcc
     3241 gagacaagca gagtggaaag acaatcctgg attttcttaa gtccgatgga tttgccaacc
     3301 ggaacttcat gcagttgatc catgatgact ctctcacctt taaggaggac atccagaaag
     3361 cacaagtttc tggccagggg gacagtcttc acgagcacat cgctaatctt gcaggtagcc
     3421 cagctatcaa aaagggaata ctgcagaccg ttaaggtcgt ggatgaactc gtcaaagtaa
     3481 tgggaaggca taagcccgag aatatcgtta tcgagatggc ccgagagaac caaactaccc
     3541 agaagggaca gaagaacagt agggaaagga tgaagaggat tgaagagggt ataaaagaac
     3601 tggggtccca aatccttaag gaacacccag ttgaaaacac ccagcttcag aatgagaagc
     3661 tctacctgta ctacctgcag aacggcaggg acatgtacgt ggatcaggaa ctggacatca
     3721 atcggctctc cgactacgac gtggctgcta tcgtgcccca gtcttttctc aaagatgatt
     3781 ctattgataa taaagtgttg acaagatccg ataaagctag agggaagagt gataacgtcc
     3841 cctcagaaga agttgtcaag aaaatgaaaa attattggcg gcagctgctg aacgccaaac
     3901 tgatcacaca acggaagttc gataatctga ctaaggctga acgaggtggc ctgtctgagt
     3961 tggataaagc cggcttcatc aaaaggcagc ttgttgagac acgccagatc accaagcacg
     4021 tggcccaaat tctcgattca cgcatgaaca ccaagtacga tgaaaatgac aaactgattc
     4081 gagaggtgaa agttattact ctgaagtcta agctggtctc agatttcaga aaggactttc
     4141 agttttataa ggtgagagag atcaacaatt accaccatgc gcatgatgcc tacctgaatg
     4201 cagtggtagg cactgcactt atcaaaaaat atcccaagct tgaatctgaa tttgtttacg
     4261 gagactataa agtgtacgat gttaggaaaa tgatcgcaaa gtctgagcag gaaataggca
     4321 aggccaccgc taagtacttc ttttacagca atattatgaa ttttttcaag accgagatta
     4381 cactggccaa tggagagatt cggaagcgac cacttatcga aacaaacgga gaaacaggag
     4441 aaatcgtgtg ggacaagggt agggatttcg cgacagtccg gaaggtcctg tccatgccgc
     4501 aggtgaacat cgttaaaaag accgaagtac agaccggagg cttctccaag gaaagtatcc
     4561 tcccgaaaag gaacagcgac aagctgatcg cacgcaaaaa agattgggac cccaagaaat
     4621 acggcggatt cgattctcct acagtcgctt acagtgtact ggttgtggcc aaagtggaga
     4681 aagggaagtc taaaaaactc aaaagcgtca aggaactgct gggcatcaca atcatggagc
     4741 gatcaagctt cgaaaaaaac cccatcgact ttctcgaggc gaaaggatat aaagaggtca
     4801 aaaaagacct catcattaag cttcccaagt actctctctt tgagcttgaa aacggccgga
     4861 aacgaatgct cgctagtgcg ggcgagctgc agaaaggtaa cgagctggca ctgccctcta
     4921 aatacgttaa tttcttgtat ctggccagcc actatgaaaa gctcaaaggg tctcccgaag
     4981 ataatgagca gaagcagctg ttcgtggaac aacacaaaca ctaccttgat gagatcatcg
     5041 agcaaataag cgaattctcc aaaagagtga tcctcgccga cgctaacctc gataaggtgc
     5101 tttctgctta caataagcac agggataagc ccatcaggga gcaggcagaa aacattatcc
     5161 acttgtttac tctgaccaac ttgggcgcgc ctgcagcctt caagtacttc gacaccacca
     5221 tagacagaaa gcggtacacc tctacaaagg aggtcctgga cgccacactg attcatcagt
     5281 caattacggg gctctatgaa acaagaatcg acctctctca gctcggtgga gacagcaggg
     5341 ctgaccccaa gaagaagagg aaggtgtgag tttaaacggg ggaggctaac tgaaacacgg
     5401 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     5461 cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt
     5521 cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc
     5581 ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc
     5641 tgccatagca gatctgcgaa atcggatccg cggccgcact gattctagct gcagaggctc
     5701 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
     5761 ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt
     5821 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt
     5881 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggatc ctcgagccac
     5941 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     6001 cggcgacgta aacggccaca agttcagcgt gaggggcgag ggcgagggcg atgccaccaa
     6061 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     6121 cctcgtgacc accctgagcc acggcgtgca gtgcttcgcc cgctaccccg accacatgaa
     6181 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     6241 cttcaaggac gacggcacct acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     6301 agtgaaccgc atcgagctga agggcgtcga cttcaaggag gacggcaaca tcctggggca
     6361 caagctggag tacaacttca acagccacaa catctatatc atggccgtca agcagaagaa
     6421 cggcatcaag gtgaacttca agatccgcca caacgtggag gacggcagcg tgcagctcgc
     6481 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacagcca
     6541 ctacctgagc acccagtccg tgctgagcaa agaccccaac gagaagcgcg atcacatggt
     6601 cctgctggag ttccgcaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     6661 agttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     6721 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     6781 atcttatcat gtctggaatt gactcaaatg atgtcaatta gtctatcaga agctcatctg
     6841 gtctcccttc cgggggacaa gacatccctg tttaatattt aaacagcagt gttcccaaac
     6901 tgggttctta tatcccttgc tctggtcaac caggttgcag ggtttcctgt cctcacagga
     6961 acgaagtccc taaagaaaca gtggcagcca ggtttagccc cggaattgac tggattcctt
     7021 ttttagggcc cattggtatg gctttttccc cgtatccccc caggtgtctg caggctcaaa
     7081 gagcagcgag aagcgttcag aggaaagcga tcccgtgcca ccttccccgt gcccgggctg
     7141 tccccgcacg ctgccggctc ggggatgcgg ggggagcgcc ggaccggagc ggagccccgg
     7201 gcggctcgct gctgccccct agcgggggag ggacgtaatt acatccctgg gggctttggg
     7261 ggggggctgt ccctgatatc tataacaaga aaatatatat ataataagtt atcacgtaag
     7321 tagaacatga aataacaata taattatcgt atgagttaaa tcttaaaagt cacgtaaaag
     7381 ataatcatgc gtcattttga ctcacgcggt cgttatagtt caaaatcagt gacacttacc
     7441 gcattgacaa gcacgcctca cgggagctcc aagcggcgac tgagatgtcc taaatgcaca
     7501 gcgacggatt cgcgctattt agaaagagag agcaatattt caagaatgca tgcgtcaatt
     7561 ttacgcagac tatctttcta gggttaatct agctgcatca ggatcatatc gtcgggtctt
     7621 ttttccggct cagtcatcgc ccaagctggc gctatctggg catcggggag gaagaagccc
     7681 gtgccttttc ccgcgaggtt gaagcggcat ggaaagagtt tgccgaggat gactgctgct
     7741 gcattgacgt tgagcgaaaa cgcacgttta ccatgatgat tcgggaaggt gtggccatgc
     7801 acgcctttaa cggtgaactg ttcgttcagg ccacctggga taccagttcg tcgcggcttt
     7861 tccggacaca gttccggatg gtcagcccga agcgcatcag caacccgaac aataccggcg
     7921 acagccggaa ctgccgtgcc ggtgtgcaga ttaatgacag cggtgcggcg ctgggatatt
     7981 acgtcagcga ggacgggtat cctggctgga tgccgcagaa atggacatgg ataccccgtg
     8041 agttacccgg cgggcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     8101 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     8161 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     8221 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     8281 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     8341 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     8401 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     8461 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     8521 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     8581 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     8641 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     8701 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     8761 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     8821 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     8881 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     8941 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     9001 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     9061 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     9121 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     9181 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     9241 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     9301 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     9361 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     9421 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     9481 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     9541 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     9601 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     9661 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     9721 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     9781 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     9841 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     9901 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     9961 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
    10021 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
    10081 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
    10141 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
    10201 cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg
    10261 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
    10321 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
    10381 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
    10441 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
    10501 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
    10561 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
    10621 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
    10681 tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
    10741 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
    10801 gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcctc gttcattcac
    10861 gtttttgaac ccgtggagga cgggcagact cgcggtgcaa atgtgtttta cagcgtgatg
    10921 gagcagatga agatgctcga cacgctgcag aacacgcagc tagattaacc ctagaaagat
    10981 aatcatattg tgacgtacgt taaagataat catgtgtaaa attgacgcat gtgttttatc
    11041 ggtctgtata tcgaggttta tttattaatt tgaatagata ttaagtttta ttatatttac
    11101 acttacatac taataataaa ttcaacaaac aatttattta tgtttattta tttattaaaa
    11161 aaaacaaaaa ctcaaaattt cttctataaa gtaacaaaac ttttatgagg gacagccccc
    11221 ccccaaagcc cccagggatg taattacgtc cctcccccgc tagggggcag cagcgagccg
    11281 cccggggctc cgctccggtc cggcgctccc cccgcatccc cgagccggca gcgtgcgggg
    11341 acagcccggg cacggggaag gtggcacggg atcgctttcc tctgaacgct tctcgctgct
    11401 ctttgagcct gcagacacct ggggggatac ggggaaaagg cctccacggc cgacattgat
    11461 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
    11521 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
    11581 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
    11641 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
    11701 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
    11761 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
    11821 ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct
    11881 ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg
    11941 gggggg
//
