LOCUS       sequence-354097-       13478 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac compatible plasmid expressing dCas9-VPR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13478)
  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 13478)
  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_Puro
FEATURES             Location/Qualifiers
     source          1..13478
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          145..1161
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1085..1107
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1169..1188
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1257..1266
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1263..1289,1290..1292,1293..3776,3777..3782,3783..3848,
                     3849..3851,3852..5366)
                     /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             5379..5399
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5424..5573
                     /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             5598..5618
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6048..6404
                     /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             6423..6992
                     /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(7017..7036)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7061..7109
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        7341..7552
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             7568..8167
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(7568..7587)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8064..8084
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    8186..8307
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8223..8242)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8277..8296
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   9070..9104
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(9595..9611)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9595..9611)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9608..9630)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9619..9635
                     /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(9643..9649,9650..9667,9668..9673))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9688..9709
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9826..9843)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9997..10585)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10077..10096)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10756..11547,11548..11616))
                     /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     11379..11398
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11617..11721)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(11747..12202)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11884..11905)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     12096..12115
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12329..12351
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     12343..12360
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     12344..12360
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   12490..12552
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        12973..13352
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga gaggtgcggc ggcagccaat
       61 cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg gcggccctat
      121 aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc
      181 tccgccgccg cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg
      241 agcgggcggg acggcccttc tcctccgggc tgtaattagc gcttggttta atgacggctt
      301 gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc gggagggccc tttgtgcggg
      361 gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg ggagcgccgc gtgcggctcc
      421 gcgctgcccg gcggctgtga gcgctgcggg cgcggcgcgg ggctttttgc gctccgcagt
      481 gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt gcgggggggg ctgcgagggg
      541 aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag cagggggtgt gggcgcgtcg
      601 gtcgggctgc aaccccccct gcacccccct ccccgagttg ctgagcacgg cccggcttcg
      661 ggtgcggggc tccgtacggg gcgtggcgcg gggctcgccg tgccgggcgg ggggtggcgg
      721 caggtggggg tgccgggcgg ggcggggccg cctcgggccg gggagggctc gggggagggg
      781 cgcggcggcc cccggagcgc cggcggctgt cgaggcgcgg cgagccgcag ccattgcctt
      841 ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt cccaaatctg tgcggagccg
      901 aaatctggga ggcgccgccg caccccctct agcgggcgcg gggcgaagcg gtgcggcgcc
      961 ggcaggaagg aaatgggcgg ggagggcctt cgtgcgtcgc cgcgccgccg tccccttctc
     1021 cctctccagc ctcggggctg tccgcggggg gacggctgcc ttcggggggg acggggcagg
     1081 gcggggttcg gcttctggcg tgtgaccggc ggctctagag cctctgctaa ccatgttcat
     1141 gccttcttct ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat
     1201 tttggcaaag aatctagata ccctcgtaaa ggtctagagc tagcgaattc gaatttgcca
     1261 ccatggacaa gaagtactcc attgggctcg ctatcggcac aaacagcgtc ggctgggccg
     1321 tcattacgga cgagtacaag gtgccgagca aaaaattcaa agttctgggc aataccgatc
     1381 gccacagcat aaagaagaac ctcattggcg ccctcctgtt cgactccggg gagacggccg
     1441 aagccacgcg gctcaaaaga acagcacggc gcagatatac ccgcagaaag aatcggatct
     1501 gctacctgca ggagatcttt agtaatgaga tggctaaggt ggatgactct ttcttccata
     1561 ggctggagga gtcctttttg gtggaggagg ataaaaagca cgagcgccac ccaatctttg
     1621 gcaatatcgt ggacgaggtg gcgtaccatg aaaagtaccc aaccatatat catctgagga
     1681 agaagcttgt agacagtact gataaggctg acttgcggtt gatctatctc gcgctggcgc
     1741 atatgatcaa atttcgggga cacttcctca tcgaggggga cctgaaccca gacaacagcg
     1801 atgtcgacaa actctttatc caactggttc agacttacaa tcagcttttc gaagagaacc
     1861 cgatcaacgc atccggagtt gacgccaaag caatcctgag cgctaggctg tccaaatccc
     1921 ggcggctcga aaacctcatc gcacagctcc ctggggagaa gaagaacggc ctgtttggta
     1981 atcttatcgc cctgtcactc gggctgaccc ccaactttaa atctaacttc gacctggccg
     2041 aagatgccaa gcttcaactg agcaaagaca cctacgatga tgatctcgac aatctgctgg
     2101 cccagatcgg cgaccagtac gcagaccttt ttttggcggc aaagaacctg tcagacgcca
     2161 ttctgctgag tgatattctg cgagtgaaca cggagatcac caaagctccg ctgagcgcta
     2221 gtatgatcaa gcgctatgat gagcaccacc aagacttgac tttgctgaag gcccttgtca
     2281 gacagcaact gcctgagaag tacaaggaaa ttttcttcga tcagtctaaa aatggctacg
     2341 ccggatacat tgacggcgga gcaagccagg aggaatttta caaatttatt aagcccatct
     2401 tggaaaaaat ggacggcacc gaggagctgc tggtaaagct taacagagaa gatctgttgc
     2461 gcaaacagcg cactttcgac aatggaagca tcccccacca gattcacctg ggcgaactgc
     2521 acgctatcct caggcggcaa gaggatttct accccttttt gaaagataac agggaaaaga
     2581 ttgagaaaat cctcacattt cggataccct actatgtagg ccccctcgcc cggggaaatt
     2641 ccagattcgc gtggatgact cgcaaatcag aagagaccat cactccctgg aacttcgagg
     2701 aagtcgtgga taagggggcc tctgcccagt ccttcatcga aaggatgact aactttgata
     2761 aaaatctgcc taacgaaaag gtgcttccta aacactctct gctgtacgag tacttcacag
     2821 tttataacga gctcaccaag gtcaaatacg tcacagaagg gatgagaaag ccagcattcc
     2881 tgtctggaga gcagaagaaa gctatcgtgg acctcctctt caagacgaac cggaaagtta
     2941 ccgtgaaaca gctcaaagaa gactatttca aaaagattga atgtttcgac tctgttgaaa
     3001 tcagcggagt ggaggatcgc ttcaacgcat ccctgggaac gtatcacgat ctcctgaaaa
     3061 tcattaaaga caaggacttc ctggacaatg aggagaacga ggacattctt gaggacattg
     3121 tcctcaccct tacgttgttt gaagataggg agatgattga agaacgcttg aaaacttacg
     3181 ctcatctctt cgacgacaaa gtcatgaaac agctcaagag gcgccgatat acaggatggg
     3241 ggcggctgtc aagaaaactg atcaatggga tccgagacaa gcagagtgga aagacaatcc
     3301 tggattttct taagtccgat ggatttgcca accggaactt catgcagttg atccatgatg
     3361 actctctcac ctttaaggag gacatccaga aagcacaagt ttctggccag ggggacagtc
     3421 ttcacgagca catcgctaat cttgcaggta gcccagctat caaaaaggga atactgcaga
     3481 ccgttaaggt cgtggatgaa ctcgtcaaag taatgggaag gcataagccc gagaatatcg
     3541 ttatcgagat ggcccgagag aaccaaacta cccagaaggg acagaagaac agtagggaaa
     3601 ggatgaagag gattgaagag ggtataaaag aactggggtc ccaaatcctt aaggaacacc
     3661 cagttgaaaa cacccagctt cagaatgaga agctctacct gtactacctg cagaacggca
     3721 gggacatgta cgtggatcag gaactggaca tcaatcggct ctccgactac gacgtggctg
     3781 ctatcgtgcc ccagtctttt ctcaaagatg attctattga taataaagtg ttgacaagat
     3841 ccgataaagc tagagggaag agtgataacg tcccctcaga agaagttgtc aagaaaatga
     3901 aaaattattg gcggcagctg ctgaacgcca aactgatcac acaacggaag ttcgataatc
     3961 tgactaaggc tgaacgaggt ggcctgtctg agttggataa agccggcttc atcaaaaggc
     4021 agcttgttga gacacgccag atcaccaagc acgtggccca aattctcgat tcacgcatga
     4081 acaccaagta cgatgaaaat gacaaactga ttcgagaggt gaaagttatt actctgaagt
     4141 ctaagctggt ctcagatttc agaaaggact ttcagtttta taaggtgaga gagatcaaca
     4201 attaccacca tgcgcatgat gcctacctga atgcagtggt aggcactgca cttatcaaaa
     4261 aatatcccaa gcttgaatct gaatttgttt acggagacta taaagtgtac gatgttagga
     4321 aaatgatcgc aaagtctgag caggaaatag gcaaggccac cgctaagtac ttcttttaca
     4381 gcaatattat gaattttttc aagaccgaga ttacactggc caatggagag attcggaagc
     4441 gaccacttat cgaaacaaac ggagaaacag gagaaatcgt gtgggacaag ggtagggatt
     4501 tcgcgacagt ccggaaggtc ctgtccatgc cgcaggtgaa catcgttaaa aagaccgaag
     4561 tacagaccgg aggcttctcc aaggaaagta tcctcccgaa aaggaacagc gacaagctga
     4621 tcgcacgcaa aaaagattgg gaccccaaga aatacggcgg attcgattct cctacagtcg
     4681 cttacagtgt actggttgtg gccaaagtgg agaaagggaa gtctaaaaaa ctcaaaagcg
     4741 tcaaggaact gctgggcatc acaatcatgg agcgatcaag cttcgaaaaa aaccccatcg
     4801 actttctcga ggcgaaagga tataaagagg tcaaaaaaga cctcatcatt aagcttccca
     4861 agtactctct ctttgagctt gaaaacggcc ggaaacgaat gctcgctagt gcgggcgagc
     4921 tgcagaaagg taacgagctg gcactgccct ctaaatacgt taatttcttg tatctggcca
     4981 gccactatga aaagctcaaa gggtctcccg aagataatga gcagaagcag ctgttcgtgg
     5041 aacaacacaa acactacctt gatgagatca tcgagcaaat aagcgaattc tccaaaagag
     5101 tgatcctcgc cgacgctaac ctcgataagg tgctttctgc ttacaataag cacagggata
     5161 agcccatcag ggagcaggca gaaaacatta tccacttgtt tactctgacc aacttgggcg
     5221 cgcctgcagc cttcaagtac ttcgacacca ccatagacag aaagcggtac acctctacaa
     5281 aggaggtcct ggacgccaca ctgattcatc agtcaattac ggggctctat gaaacaagaa
     5341 tcgacctctc tcagctcggt ggagacagca gggctgaccc caagaagaag aggaaggtgg
     5401 aggccagcgg ttccggacgg gctgacgcat tggacgattt tgatctggat atgctgggaa
     5461 gtgacgccct cgatgatttt gaccttgaca tgcttggttc ggatgccctt gatgactttg
     5521 acctcgacat gctcggcagt gacgcccttg atgatttcga cctggacatg ctgattaact
     5581 ctagaagttc cggatctccg aaaaagaaac gcaaagttgg tagccagtac ctgcccgaca
     5641 ccgacgaccg gcaccggatc gaggaaaagc ggaagcggac ctacgagaca ttcaagagca
     5701 tcatgaagaa gtcccccttc agcggcccca ccgaccctag acctccacct agaagaatcg
     5761 ccgtgcccag cagatccagc gccagcgtgc caaaacctgc cccccagcct taccccttca
     5821 ccagcagcct gagcaccatc aactacgacg agttccctac catggtgttc cccagcggcc
     5881 agatctctca ggcctctgct ctggctccag cccctcctca ggtgctgcct caggctcctg
     5941 ctcctgcacc agctccagcc atggtgtctg cactggctca ggcaccagca cccgtgcctg
     6001 tgctggctcc tggacctcca caggctgtgg ctccaccagc ccctaaacct acacaggccg
     6061 gcgagggcac actgtctgaa gctctgctgc agctgcagtt cgacgacgag gatctgggag
     6121 ccctgctggg aaacagcacc gatcctgccg tgttcaccga cctggccagc gtggacaaca
     6181 gcgagttcca gcagctgctg aaccagggca tccctgtggc ccctcacacc accgagccca
     6241 tgctgatgga ataccccgag gccatcaccc ggctcgtgac aggcgctcag aggcctcctg
     6301 atccagctcc tgcccctctg ggagcaccag gcctgcctaa tggactgctg tctggcgacg
     6361 aggacttcag ctctatcgcc gacatggact tctccgcact gctgggtagc ggatcgggat
     6421 ctcgggattc cagggaaggg atgtttttgc cgaagcctga ggccggctcc gctattagtg
     6481 acgtgtttga gggccgcgag gtgtgccagc caaaacgaat ccggccattt catcctccag
     6541 gaagtccatg ggccaaccgc ccactccccg ccagcctcgc accaacacca accggtccag
     6601 tacatgagcc agtcgggtca ctgaccccgg caccagtccc tcagccactg gatccagcgc
     6661 ccgcagtgac tcccgaggcc agtcacctgt tggaggatcc cgatgaagag acgagccagg
     6721 ctgtcaaagc ccttcgggag atggccgata ctgtgattcc ccagaaggaa gaggctgcaa
     6781 tctgtggcca aatggacctt tcccatccgc ccccaagggg ccatctggat gagctgacaa
     6841 ccacacttga gtccatgacc gaggatctga acctggactc acccctgacc ccggaattga
     6901 acgagattct ggataccttc ctgaacgacg agtgcctctt gcatgccatg catatcagca
     6961 caggactgtc catcttcgac acatctctgt tttgagttta aacgggggag gctaactgaa
     7021 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     7081 aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     7141 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     7201 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     7261 aggccctgcc atagcagatc tgcgaaatcg gatccgcggc cgcactgatt ctagctgcag
     7321 aggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
     7381 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
     7441 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
     7501 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggatcctcg
     7561 agccaccatg accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtcccccg
     7621 ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga
     7681 cccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg
     7741 gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac
     7801 gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt
     7861 gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc
     7921 caaggagccc gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg
     7981 tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc
     8041 cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt
     8101 caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg
     8161 tgcctgaatc tagtgtcgac aagttaactt gtttattgca gcttataatg gttacaaata
     8221 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     8281 tttgtccaaa ctcatcaatg tatcttatca tgtctggaat tgactcaaat gatgtcaatt
     8341 agtctatcag aagctcatct ggtctccctt ccgggggaca agacatccct gtttaatatt
     8401 taaacagcag tgttcccaaa ctgggttctt atatcccttg ctctggtcaa ccaggttgca
     8461 gggtttcctg tcctcacagg aacgaagtcc ctaaagaaac agtggcagcc aggtttagcc
     8521 ccggaattga ctggattcct tttttagggc ccattggtat ggctttttcc ccgtatcccc
     8581 ccaggtgtct gcaggctcaa agagcagcga gaagcgttca gaggaaagcg atcccgtgcc
     8641 accttccccg tgcccgggct gtccccgcac gctgccggct cggggatgcg gggggagcgc
     8701 cggaccggag cggagccccg ggcggctcgc tgctgccccc tagcggggga gggacgtaat
     8761 tacatccctg ggggctttgg gggggggctg tccctgatat ctataacaag aaaatatata
     8821 tataataagt tatcacgtaa gtagaacatg aaataacaat ataattatcg tatgagttaa
     8881 atcttaaaag tcacgtaaaa gataatcatg cgtcattttg actcacgcgg tcgttatagt
     8941 tcaaaatcag tgacacttac cgcattgaca agcacgcctc acgggagctc caagcggcga
     9001 ctgagatgtc ctaaatgcac agcgacggat tcgcgctatt tagaaagaga gagcaatatt
     9061 tcaagaatgc atgcgtcaat tttacgcaga ctatctttct agggttaatc tagctgcatc
     9121 aggatcatat cgtcgggtct tttttccggc tcagtcatcg cccaagctgg cgctatctgg
     9181 gcatcgggga ggaagaagcc cgtgcctttt cccgcgaggt tgaagcggca tggaaagagt
     9241 ttgccgagga tgactgctgc tgcattgacg ttgagcgaaa acgcacgttt accatgatga
     9301 ttcgggaagg tgtggccatg cacgccttta acggtgaact gttcgttcag gccacctggg
     9361 ataccagttc gtcgcggctt ttccggacac agttccggat ggtcagcccg aagcgcatca
     9421 gcaacccgaa caataccggc gacagccgga actgccgtgc cggtgtgcag attaatgaca
     9481 gcggtgcggc gctgggatat tacgtcagcg aggacgggta tcctggctgg atgccgcaga
     9541 aatggacatg gataccccgt gagttacccg gcgggcgcgc ttggcgtaat catggtcata
     9601 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     9661 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     9721 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     9781 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     9841 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     9901 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     9961 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
    10021 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
    10081 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
    10141 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
    10201 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
    10261 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
    10321 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
    10381 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
    10441 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
    10501 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
    10561 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
    10621 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
    10681 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
    10741 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
    10801 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
    10861 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
    10921 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
    10981 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
    11041 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
    11101 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
    11161 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
    11221 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
    11281 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
    11341 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
    11401 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
    11461 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
    11521 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
    11581 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
    11641 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
    11701 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt
    11761 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
    11821 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
    11881 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
    11941 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
    12001 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
    12061 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
    12121 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
    12181 aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg
    12241 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
    12301 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
    12361 gagcgcgcct cgttcattca cgtttttgaa cccgtggagg acgggcagac tcgcggtgca
    12421 aatgtgtttt acagcgtgat ggagcagatg aagatgctcg acacgctgca gaacacgcag
    12481 ctagattaac cctagaaaga taatcatatt gtgacgtacg ttaaagataa tcatgtgtaa
    12541 aattgacgca tgtgttttat cggtctgtat atcgaggttt atttattaat ttgaatagat
    12601 attaagtttt attatattta cacttacata ctaataataa attcaacaaa caatttattt
    12661 atgtttattt atttattaaa aaaaacaaaa actcaaaatt tcttctataa agtaacaaaa
    12721 cttttatgag ggacagcccc cccccaaagc ccccagggat gtaattacgt ccctcccccg
    12781 ctagggggca gcagcgagcc gcccggggct ccgctccggt ccggcgctcc ccccgcatcc
    12841 ccgagccggc agcgtgcggg gacagcccgg gcacggggaa ggtggcacgg gatcgctttc
    12901 ctctgaacgc ttctcgctgc tctttgagcc tgcagacacc tggggggata cggggaaaag
    12961 gcctccacgg ccgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
    13021 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
    13081 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
    13141 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
    13201 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
    13261 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
    13321 acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg ttctgcttca
    13381 ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat
    13441 tttgtgcagc gatgggggcg gggggggggg gggggcgc
//
