LOCUS       sequence-371889-       13766 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  PiggyBac compatible plasmid expressing dCas9-VPR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 13766)
  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 13766)
  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_VPR_mcherry
FEATURES             Location/Qualifiers
     source          1..13766
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     intron          221..1237
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1161..1183
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1245..1264
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1333..1342
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1339..1365,1366..1368,1369..3852,3853..3858,3859..3924,
                     3925..3927,3928..5442)
                     /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             5455..5475
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5500..5649
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             5674..5694
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6124..6480
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALL"
     CDS             6499..7068
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
     primer_bind     complement(7093..7112)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7137..7185
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        7417..7628
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      7638..7647
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(7762..7780)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     8030..8049
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    8331..8452
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8368..8387)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8422..8441
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   9215..9249
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(9740..9756)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9740..9756)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9753..9775)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9764..9780
                     /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(9788..9794,9795..9812,9813..9818))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9833..9854
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9971..9988)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10142..10730)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10222..10241)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10901..11692,11693..11761))
                     /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     11524..11543
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11762..11866)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(11892..12347)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(12029..12050)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     12241..12260
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12474..12496
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     12488..12505
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     12489..12505
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   12635..12697
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        13118..13497
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 anktttgktt ttwktnwttk tttkawkwwk ktgggggggg gggggggggg gggggggggg
       61 ggggcgcgcg csaggcgggg cggggcgggg cgaggggcgg ggcggggcga ggcggagagg
      121 tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg
      181 gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct
      241 tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc
      301 gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt
      361 ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa agccttgagg ggctccggga
      421 gggccctttg tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag
      481 cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct
      541 ttttgcgctc cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc cgcggtgcgg
      601 ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg ggtgagcagg
      661 gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc gagttgctga
      721 gcacggcccg gcttcgggtg cggggctccg tacggggcgt ggcgcggggc tcgccgtgcc
      781 gggcgggggg tggcggcagg tgggggtgcc gggcggggcg gggccgcctc gggccgggga
      841 gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag
      901 ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc ctttgtccca
      961 aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc
     1021 gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg
     1081 ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg gctgccttcg
     1141 ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc
     1201 tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat
     1261 tgtgctgtct catcattttg gcaaagaatc tagataccct cgtaaaggtc tagagctagc
     1321 gaattcgaat ttgccaccat ggacaagaag tactccattg ggctcgctat cggcacaaac
     1381 agcgtcggct gggccgtcat tacggacgag tacaaggtgc cgagcaaaaa attcaaagtt
     1441 ctgggcaata ccgatcgcca cagcataaag aagaacctca ttggcgccct cctgttcgac
     1501 tccggggaga cggccgaagc cacgcggctc aaaagaacag cacggcgcag atatacccgc
     1561 agaaagaatc ggatctgcta cctgcaggag atctttagta atgagatggc taaggtggat
     1621 gactctttct tccataggct ggaggagtcc tttttggtgg aggaggataa aaagcacgag
     1681 cgccacccaa tctttggcaa tatcgtggac gaggtggcgt accatgaaaa gtacccaacc
     1741 atatatcatc tgaggaagaa gcttgtagac agtactgata aggctgactt gcggttgatc
     1801 tatctcgcgc tggcgcatat gatcaaattt cggggacact tcctcatcga gggggacctg
     1861 aacccagaca acagcgatgt cgacaaactc tttatccaac tggttcagac ttacaatcag
     1921 cttttcgaag agaacccgat caacgcatcc ggagttgacg ccaaagcaat cctgagcgct
     1981 aggctgtcca aatcccggcg gctcgaaaac ctcatcgcac agctccctgg ggagaagaag
     2041 aacggcctgt ttggtaatct tatcgccctg tcactcgggc tgacccccaa ctttaaatct
     2101 aacttcgacc tggccgaaga tgccaagctt caactgagca aagacaccta cgatgatgat
     2161 ctcgacaatc tgctggccca gatcggcgac cagtacgcag accttttttt ggcggcaaag
     2221 aacctgtcag acgccattct gctgagtgat attctgcgag tgaacacgga gatcaccaaa
     2281 gctccgctga gcgctagtat gatcaagcgc tatgatgagc accaccaaga cttgactttg
     2341 ctgaaggccc ttgtcagaca gcaactgcct gagaagtaca aggaaatttt cttcgatcag
     2401 tctaaaaatg gctacgccgg atacattgac ggcggagcaa gccaggagga attttacaaa
     2461 tttattaagc ccatcttgga aaaaatggac ggcaccgagg agctgctggt aaagcttaac
     2521 agagaagatc tgttgcgcaa acagcgcact ttcgacaatg gaagcatccc ccaccagatt
     2581 cacctgggcg aactgcacgc tatcctcagg cggcaagagg atttctaccc ctttttgaaa
     2641 gataacaggg aaaagattga gaaaatcctc acatttcgga taccctacta tgtaggcccc
     2701 ctcgcccggg gaaattccag attcgcgtgg atgactcgca aatcagaaga gaccatcact
     2761 ccctggaact tcgaggaagt cgtggataag ggggcctctg cccagtcctt catcgaaagg
     2821 atgactaact ttgataaaaa tctgcctaac gaaaaggtgc ttcctaaaca ctctctgctg
     2881 tacgagtact tcacagttta taacgagctc accaaggtca aatacgtcac agaagggatg
     2941 agaaagccag cattcctgtc tggagagcag aagaaagcta tcgtggacct cctcttcaag
     3001 acgaaccgga aagttaccgt gaaacagctc aaagaagact atttcaaaaa gattgaatgt
     3061 ttcgactctg ttgaaatcag cggagtggag gatcgcttca acgcatccct gggaacgtat
     3121 cacgatctcc tgaaaatcat taaagacaag gacttcctgg acaatgagga gaacgaggac
     3181 attcttgagg acattgtcct cacccttacg ttgtttgaag atagggagat gattgaagaa
     3241 cgcttgaaaa cttacgctca tctcttcgac gacaaagtca tgaaacagct caagaggcgc
     3301 cgatatacag gatgggggcg gctgtcaaga aaactgatca atgggatccg agacaagcag
     3361 agtggaaaga caatcctgga ttttcttaag tccgatggat ttgccaaccg gaacttcatg
     3421 cagttgatcc atgatgactc tctcaccttt aaggaggaca tccagaaagc acaagtttct
     3481 ggccaggggg acagtcttca cgagcacatc gctaatcttg caggtagccc agctatcaaa
     3541 aagggaatac tgcagaccgt taaggtcgtg gatgaactcg tcaaagtaat gggaaggcat
     3601 aagcccgaga atatcgttat cgagatggcc cgagagaacc aaactaccca gaagggacag
     3661 aagaacagta gggaaaggat gaagaggatt gaagagggta taaaagaact ggggtcccaa
     3721 atccttaagg aacacccagt tgaaaacacc cagcttcaga atgagaagct ctacctgtac
     3781 tacctgcaga acggcaggga catgtacgtg gatcaggaac tggacatcaa tcggctctcc
     3841 gactacgacg tggctgctat cgtgccccag tcttttctca aagatgattc tattgataat
     3901 aaagtgttga caagatccga taaagctaga gggaagagtg ataacgtccc ctcagaagaa
     3961 gttgtcaaga aaatgaaaaa ttattggcgg cagctgctga acgccaaact gatcacacaa
     4021 cggaagttcg ataatctgac taaggctgaa cgaggtggcc tgtctgagtt ggataaagcc
     4081 ggcttcatca aaaggcagct tgttgagaca cgccagatca ccaagcacgt ggcccaaatt
     4141 ctcgattcac gcatgaacac caagtacgat gaaaatgaca aactgattcg agaggtgaaa
     4201 gttattactc tgaagtctaa gctggtctca gatttcagaa aggactttca gttttataag
     4261 gtgagagaga tcaacaatta ccaccatgcg catgatgcct acctgaatgc agtggtaggc
     4321 actgcactta tcaaaaaata tcccaagctt gaatctgaat ttgtttacgg agactataaa
     4381 gtgtacgatg ttaggaaaat gatcgcaaag tctgagcagg aaataggcaa ggccaccgct
     4441 aagtacttct tttacagcaa tattatgaat tttttcaaga ccgagattac actggccaat
     4501 ggagagattc ggaagcgacc acttatcgaa acaaacggag aaacaggaga aatcgtgtgg
     4561 gacaagggta gggatttcgc gacagtccgg aaggtcctgt ccatgccgca ggtgaacatc
     4621 gttaaaaaga ccgaagtaca gaccggaggc ttctccaagg aaagtatcct cccgaaaagg
     4681 aacagcgaca agctgatcgc acgcaaaaaa gattgggacc ccaagaaata cggcggattc
     4741 gattctccta cagtcgctta cagtgtactg gttgtggcca aagtggagaa agggaagtct
     4801 aaaaaactca aaagcgtcaa ggaactgctg ggcatcacaa tcatggagcg atcaagcttc
     4861 gaaaaaaacc ccatcgactt tctcgaggcg aaaggatata aagaggtcaa aaaagacctc
     4921 atcattaagc ttcccaagta ctctctcttt gagcttgaaa acggccggaa acgaatgctc
     4981 gctagtgcgg gcgagctgca gaaaggtaac gagctggcac tgccctctaa atacgttaat
     5041 ttcttgtatc tggccagcca ctatgaaaag ctcaaagggt ctcccgaaga taatgagcag
     5101 aagcagctgt tcgtggaaca acacaaacac taccttgatg agatcatcga gcaaataagc
     5161 gaattctcca aaagagtgat cctcgccgac gctaacctcg ataaggtgct ttctgcttac
     5221 aataagcaca gggataagcc catcagggag caggcagaaa acattatcca cttgtttact
     5281 ctgaccaact tgggcgcgcc tgcagccttc aagtacttcg acaccaccat agacagaaag
     5341 cggtacacct ctacaaagga ggtcctggac gccacactga ttcatcagtc aattacgggg
     5401 ctctatgaaa caagaatcga cctctctcag ctcggtggag acagcagggc tgaccccaag
     5461 aagaagagga aggtggaggc cagcggttcc ggacgggctg acgcattgga cgattttgat
     5521 ctggatatgc tgggaagtga cgccctcgat gattttgacc ttgacatgct tggttcggat
     5581 gcccttgatg actttgacct cgacatgctc ggcagtgacg cccttgatga tttcgacctg
     5641 gacatgctga ttaactctag aagttccgga tctccgaaaa agaaacgcaa agttggtagc
     5701 cagtacctgc ccgacaccga cgaccggcac cggatcgagg aaaagcggaa gcggacctac
     5761 gagacattca agagcatcat gaagaagtcc cccttcagcg gccccaccga ccctagacct
     5821 ccacctagaa gaatcgccgt gcccagcaga tccagcgcca gcgtgccaaa acctgccccc
     5881 cagccttacc ccttcaccag cagcctgagc accatcaact acgacgagtt ccctaccatg
     5941 gtgttcccca gcggccagat ctctcaggcc tctgctctgg ctccagcccc tcctcaggtg
     6001 ctgcctcagg ctcctgctcc tgcaccagct ccagccatgg tgtctgcact ggctcaggca
     6061 ccagcacccg tgcctgtgct ggctcctgga cctccacagg ctgtggctcc accagcccct
     6121 aaacctacac aggccggcga gggcacactg tctgaagctc tgctgcagct gcagttcgac
     6181 gacgaggatc tgggagccct gctgggaaac agcaccgatc ctgccgtgtt caccgacctg
     6241 gccagcgtgg acaacagcga gttccagcag ctgctgaacc agggcatccc tgtggcccct
     6301 cacaccaccg agcccatgct gatggaatac cccgaggcca tcacccggct cgtgacaggc
     6361 gctcagaggc ctcctgatcc agctcctgcc cctctgggag caccaggcct gcctaatgga
     6421 ctgctgtctg gcgacgagga cttcagctct atcgccgaca tggacttctc cgcactgctg
     6481 ggtagcggat cgggatctcg ggattccagg gaagggatgt ttttgccgaa gcctgaggcc
     6541 ggctccgcta ttagtgacgt gtttgagggc cgcgaggtgt gccagccaaa acgaatccgg
     6601 ccatttcatc ctccaggaag tccatgggcc aaccgcccac tccccgccag cctcgcacca
     6661 acaccaaccg gtccagtaca tgagccagtc gggtcactga ccccggcacc agtccctcag
     6721 ccactggatc cagcgcccgc agtgactccc gaggccagtc acctgttgga ggatcccgat
     6781 gaagagacga gccaggctgt caaagccctt cgggagatgg ccgatactgt gattccccag
     6841 aaggaagagg ctgcaatctg tggccaaatg gacctttccc atccgccccc aaggggccat
     6901 ctggatgagc tgacaaccac acttgagtcc atgaccgagg atctgaacct ggactcaccc
     6961 ctgaccccgg aattgaacga gattctggat accttcctga acgacgagtg cctcttgcat
     7021 gccatgcata tcagcacagg actgtccatc ttcgacacat ctctgttttg agtttaaacg
     7081 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     7141 aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc
     7201 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     7261 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     7321 ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg aaatcggatc cgcggccgca
     7381 ctgattctag ctgcagaggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag
     7441 tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg
     7501 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag
     7561 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca
     7621 gaacacagga tcctcgagcc accatggcca tcatcaagga gttcatgcgc ttcaaggtgc
     7681 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     7741 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
     7801 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
     7861 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     7921 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg
     7981 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
     8041 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
     8101 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
     8161 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
     8221 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
     8281 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaagtt aacttgttta
     8341 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     8401 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     8461 ggaattgact caaatgatgt caattagtct atcagaagct catctggtct cccttccggg
     8521 ggacaagaca tccctgttta atatttaaac agcagtgttc ccaaactggg ttcttatatc
     8581 ccttgctctg gtcaaccagg ttgcagggtt tcctgtcctc acaggaacga agtccctaaa
     8641 gaaacagtgg cagccaggtt tagccccgga attgactgga ttcctttttt agggcccatt
     8701 ggtatggctt tttccccgta tccccccagg tgtctgcagg ctcaaagagc agcgagaagc
     8761 gttcagagga aagcgatccc gtgccacctt ccccgtgccc gggctgtccc cgcacgctgc
     8821 cggctcgggg atgcgggggg agcgccggac cggagcggag ccccgggcgg ctcgctgctg
     8881 ccccctagcg ggggagggac gtaattacat ccctgggggc tttggggggg ggctgtccct
     8941 gatatctata acaagaaaat atatatataa taagttatca cgtaagtaga acatgaaata
     9001 acaatataat tatcgtatga gttaaatctt aaaagtcacg taaaagataa tcatgcgtca
     9061 ttttgactca cgcggtcgtt atagttcaaa atcagtgaca cttaccgcat tgacaagcac
     9121 gcctcacggg agctccaagc ggcgactgag atgtcctaaa tgcacagcga cggattcgcg
     9181 ctatttagaa agagagagca atatttcaag aatgcatgcg tcaattttac gcagactatc
     9241 tttctagggt taatctagct gcatcaggat catatcgtcg ggtctttttt ccggctcagt
     9301 catcgcccaa gctggcgcta tctgggcatc ggggaggaag aagcccgtgc cttttcccgc
     9361 gaggttgaag cggcatggaa agagtttgcc gaggatgact gctgctgcat tgacgttgag
     9421 cgaaaacgca cgtttaccat gatgattcgg gaaggtgtgg ccatgcacgc ctttaacggt
     9481 gaactgttcg ttcaggccac ctgggatacc agttcgtcgc ggcttttccg gacacagttc
     9541 cggatggtca gcccgaagcg catcagcaac ccgaacaata ccggcgacag ccggaactgc
     9601 cgtgccggtg tgcagattaa tgacagcggt gcggcgctgg gatattacgt cagcgaggac
     9661 gggtatcctg gctggatgcc gcagaaatgg acatggatac cccgtgagtt acccggcggg
     9721 cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     9781 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     9841 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     9901 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     9961 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
    10021 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
    10081 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
    10141 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
    10201 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
    10261 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
    10321 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
    10381 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
    10441 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
    10501 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
    10561 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
    10621 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
    10681 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
    10741 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
    10801 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
    10861 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
    10921 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
    10981 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
    11041 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
    11101 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
    11161 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
    11221 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
    11281 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
    11341 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
    11401 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
    11461 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
    11521 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
    11581 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
    11641 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
    11701 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
    11761 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
    11821 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
    11881 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
    11941 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
    12001 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
    12061 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
    12121 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
    12181 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
    12241 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
    12301 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
    12361 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
    12421 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
    12481 acgacgttgt aaaacgacgg ccagtgagcg cgcctcgttc attcacgttt ttgaacccgt
    12541 ggaggacggg cagactcgcg gtgcaaatgt gttttacagc gtgatggagc agatgaagat
    12601 gctcgacacg ctgcagaaca cgcagctaga ttaaccctag aaagataatc atattgtgac
    12661 gtacgttaaa gataatcatg tgtaaaattg acgcatgtgt tttatcggtc tgtatatcga
    12721 ggtttattta ttaatttgaa tagatattaa gttttattat atttacactt acatactaat
    12781 aataaattca acaaacaatt tatttatgtt tatttattta ttaaaaaaaa caaaaactca
    12841 aaatttcttc tataaagtaa caaaactttt atgagggaca gccccccccc aaagccccca
    12901 gggatgtaat tacgtccctc ccccgctagg gggcagcagc gagccgcccg gggctccgct
    12961 ccggtccggc gctccccccg catccccgag ccggcagcgt gcggggacag cccgggcacg
    13021 gggaaggtgg cacgggatcg ctttcctctg aacgcttctc gctgctcttt gagcctgcag
    13081 acacctgggg ggatacgggg aaaaggcctc cacggccgac attgattatt gactagttat
    13141 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
    13201 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
    13261 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
    13321 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
    13381 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
    13441 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc
    13501 gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt
    13561 ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg gggggggggg
    13621 ggsggcccsg gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg
    13681 ggggggggsg nngggggggg gggggggggg gggggggggg gggggggggg gggggggggg
    13741 gggggggggg gggggggggg gggggg
//
