LOCUS       sequence-354096-       12891 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 12891)
  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 12891)
  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_KanR neo
FEATURES             Location/Qualifiers
     source          1..12891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     321..343
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     405..424
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      493..502
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(499..525,526..528,529..3012,3013..3018,3019..3084,
                     3085..3087,3088..4602)
                     /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             4615..4635
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4660..4809
                     /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             4834..4854
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5284..5640
                     /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             5659..6228
                     /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(6253..6272)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6297..6345
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        6577..6788
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             6804..7598
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6804..7598
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6858..6877)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7468..7487
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7602..7723
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7639..7658)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7693..7712
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   8486..8520
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(9011..9027)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9011..9027)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9024..9046)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9035..9051
                     /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(9059..9065,9066..9083,9084..9089))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9104..9125
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9242..9259)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9413..10001)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9493..9512)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10172..10963,10964..11032))
                     /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     10795..10814
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11033..11137)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(11163..11618)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11300..11321)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     11512..11531
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11745..11767
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     11759..11776
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     11760..11776
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   11906..11968
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     enhancer        12389..12768
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag
       61 ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc ctttgtccca
      121 aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc
      181 gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg
      241 ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg gctgccttcg
      301 ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc
      361 tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat
      421 tgtgctgtct catcattttg gcaaagaatc tagataccct cgtaaaggtc tagagctagc
      481 gaattcgaat ttgccaccat ggacaagaag tactccattg ggctcgctat cggcacaaac
      541 agcgtcggct gggccgtcat tacggacgag tacaaggtgc cgagcaaaaa attcaaagtt
      601 ctgggcaata ccgatcgcca cagcataaag aagaacctca ttggcgccct cctgttcgac
      661 tccggggaga cggccgaagc cacgcggctc aaaagaacag cacggcgcag atatacccgc
      721 agaaagaatc ggatctgcta cctgcaggag atctttagta atgagatggc taaggtggat
      781 gactctttct tccataggct ggaggagtcc tttttggtgg aggaggataa aaagcacgag
      841 cgccacccaa tctttggcaa tatcgtggac gaggtggcgt accatgaaaa gtacccaacc
      901 atatatcatc tgaggaagaa gcttgtagac agtactgata aggctgactt gcggttgatc
      961 tatctcgcgc tggcgcatat gatcaaattt cggggacact tcctcatcga gggggacctg
     1021 aacccagaca acagcgatgt cgacaaactc tttatccaac tggttcagac ttacaatcag
     1081 cttttcgaag agaacccgat caacgcatcc ggagttgacg ccaaagcaat cctgagcgct
     1141 aggctgtcca aatcccggcg gctcgaaaac ctcatcgcac agctccctgg ggagaagaag
     1201 aacggcctgt ttggtaatct tatcgccctg tcactcgggc tgacccccaa ctttaaatct
     1261 aacttcgacc tggccgaaga tgccaagctt caactgagca aagacaccta cgatgatgat
     1321 ctcgacaatc tgctggccca gatcggcgac cagtacgcag accttttttt ggcggcaaag
     1381 aacctgtcag acgccattct gctgagtgat attctgcgag tgaacacgga gatcaccaaa
     1441 gctccgctga gcgctagtat gatcaagcgc tatgatgagc accaccaaga cttgactttg
     1501 ctgaaggccc ttgtcagaca gcaactgcct gagaagtaca aggaaatttt cttcgatcag
     1561 tctaaaaatg gctacgccgg atacattgac ggcggagcaa gccaggagga attttacaaa
     1621 tttattaagc ccatcttgga aaaaatggac ggcaccgagg agctgctggt aaagcttaac
     1681 agagaagatc tgttgcgcaa acagcgcact ttcgacaatg gaagcatccc ccaccagatt
     1741 cacctgggcg aactgcacgc tatcctcagg cggcaagagg atttctaccc ctttttgaaa
     1801 gataacaggg aaaagattga gaaaatcctc acatttcgga taccctacta tgtaggcccc
     1861 ctcgcccggg gaaattccag attcgcgtgg atgactcgca aatcagaaga gaccatcact
     1921 ccctggaact tcgaggaagt cgtggataag ggggcctctg cccagtcctt catcgaaagg
     1981 atgactaact ttgataaaaa tctgcctaac gaaaaggtgc ttcctaaaca ctctctgctg
     2041 tacgagtact tcacagttta taacgagctc accaaggtca aatacgtcac agaagggatg
     2101 agaaagccag cattcctgtc tggagagcag aagaaagcta tcgtggacct cctcttcaag
     2161 acgaaccgga aagttaccgt gaaacagctc aaagaagact atttcaaaaa gattgaatgt
     2221 ttcgactctg ttgaaatcag cggagtggag gatcgcttca acgcatccct gggaacgtat
     2281 cacgatctcc tgaaaatcat taaagacaag gacttcctgg acaatgagga gaacgaggac
     2341 attcttgagg acattgtcct cacccttacg ttgtttgaag atagggagat gattgaagaa
     2401 cgcttgaaaa cttacgctca tctcttcgac gacaaagtca tgaaacagct caagaggcgc
     2461 cgatatacag gatgggggcg gctgtcaaga aaactgatca atgggatccg agacaagcag
     2521 agtggaaaga caatcctgga ttttcttaag tccgatggat ttgccaaccg gaacttcatg
     2581 cagttgatcc atgatgactc tctcaccttt aaggaggaca tccagaaagc acaagtttct
     2641 ggccaggggg acagtcttca cgagcacatc gctaatcttg caggtagccc agctatcaaa
     2701 aagggaatac tgcagaccgt taaggtcgtg gatgaactcg tcaaagtaat gggaaggcat
     2761 aagcccgaga atatcgttat cgagatggcc cgagagaacc aaactaccca gaagggacag
     2821 aagaacagta gggaaaggat gaagaggatt gaagagggta taaaagaact ggggtcccaa
     2881 atccttaagg aacacccagt tgaaaacacc cagcttcaga atgagaagct ctacctgtac
     2941 tacctgcaga acggcaggga catgtacgtg gatcaggaac tggacatcaa tcggctctcc
     3001 gactacgacg tggctgctat cgtgccccag tcttttctca aagatgattc tattgataat
     3061 aaagtgttga caagatccga taaagctaga gggaagagtg ataacgtccc ctcagaagaa
     3121 gttgtcaaga aaatgaaaaa ttattggcgg cagctgctga acgccaaact gatcacacaa
     3181 cggaagttcg ataatctgac taaggctgaa cgaggtggcc tgtctgagtt ggataaagcc
     3241 ggcttcatca aaaggcagct tgttgagaca cgccagatca ccaagcacgt ggcccaaatt
     3301 ctcgattcac gcatgaacac caagtacgat gaaaatgaca aactgattcg agaggtgaaa
     3361 gttattactc tgaagtctaa gctggtctca gatttcagaa aggactttca gttttataag
     3421 gtgagagaga tcaacaatta ccaccatgcg catgatgcct acctgaatgc agtggtaggc
     3481 actgcactta tcaaaaaata tcccaagctt gaatctgaat ttgtttacgg agactataaa
     3541 gtgtacgatg ttaggaaaat gatcgcaaag tctgagcagg aaataggcaa ggccaccgct
     3601 aagtacttct tttacagcaa tattatgaat tttttcaaga ccgagattac actggccaat
     3661 ggagagattc ggaagcgacc acttatcgaa acaaacggag aaacaggaga aatcgtgtgg
     3721 gacaagggta gggatttcgc gacagtccgg aaggtcctgt ccatgccgca ggtgaacatc
     3781 gttaaaaaga ccgaagtaca gaccggaggc ttctccaagg aaagtatcct cccgaaaagg
     3841 aacagcgaca agctgatcgc acgcaaaaaa gattgggacc ccaagaaata cggcggattc
     3901 gattctccta cagtcgctta cagtgtactg gttgtggcca aagtggagaa agggaagtct
     3961 aaaaaactca aaagcgtcaa ggaactgctg ggcatcacaa tcatggagcg atcaagcttc
     4021 gaaaaaaacc ccatcgactt tctcgaggcg aaaggatata aagaggtcaa aaaagacctc
     4081 atcattaagc ttcccaagta ctctctcttt gagcttgaaa acggccggaa acgaatgctc
     4141 gctagtgcgg gcgagctgca gaaaggtaac gagctggcac tgccctctaa atacgttaat
     4201 ttcttgtatc tggccagcca ctatgaaaag ctcaaagggt ctcccgaaga taatgagcag
     4261 aagcagctgt tcgtggaaca acacaaacac taccttgatg agatcatcga gcaaataagc
     4321 gaattctcca aaagagtgat cctcgccgac gctaacctcg ataaggtgct ttctgcttac
     4381 aataagcaca gggataagcc catcagggag caggcagaaa acattatcca cttgtttact
     4441 ctgaccaact tgggcgcgcc tgcagccttc aagtacttcg acaccaccat agacagaaag
     4501 cggtacacct ctacaaagga ggtcctggac gccacactga ttcatcagtc aattacgggg
     4561 ctctatgaaa caagaatcga cctctctcag ctcggtggag acagcagggc tgaccccaag
     4621 aagaagagga aggtggaggc cagcggttcc ggacgggctg acgcattgga cgattttgat
     4681 ctggatatgc tgggaagtga cgccctcgat gattttgacc ttgacatgct tggttcggat
     4741 gcccttgatg actttgacct cgacatgctc ggcagtgacg cccttgatga tttcgacctg
     4801 gacatgctga ttaactctag aagttccgga tctccgaaaa agaaacgcaa agttggtagc
     4861 cagtacctgc ccgacaccga cgaccggcac cggatcgagg aaaagcggaa gcggacctac
     4921 gagacattca agagcatcat gaagaagtcc cccttcagcg gccccaccga ccctagacct
     4981 ccacctagaa gaatcgccgt gcccagcaga tccagcgcca gcgtgccaaa acctgccccc
     5041 cagccttacc ccttcaccag cagcctgagc accatcaact acgacgagtt ccctaccatg
     5101 gtgttcccca gcggccagat ctctcaggcc tctgctctgg ctccagcccc tcctcaggtg
     5161 ctgcctcagg ctcctgctcc tgcaccagct ccagccatgg tgtctgcact ggctcaggca
     5221 ccagcacccg tgcctgtgct ggctcctgga cctccacagg ctgtggctcc accagcccct
     5281 aaacctacac aggccggcga gggcacactg tctgaagctc tgctgcagct gcagttcgac
     5341 gacgaggatc tgggagccct gctgggaaac agcaccgatc ctgccgtgtt caccgacctg
     5401 gccagcgtgg acaacagcga gttccagcag ctgctgaacc agggcatccc tgtggcccct
     5461 cacaccaccg agcccatgct gatggaatac cccgaggcca tcacccggct cgtgacaggc
     5521 gctcagaggc ctcctgatcc agctcctgcc cctctgggag caccaggcct gcctaatgga
     5581 ctgctgtctg gcgacgagga cttcagctct atcgccgaca tggacttctc cgcactgctg
     5641 ggtagcggat cgggatctcg ggattccagg gaagggatgt ttttgccgaa gcctgaggcc
     5701 ggctccgcta ttagtgacgt gtttgagggc cgcgaggtgt gccagccaaa acgaatccgg
     5761 ccatttcatc ctccaggaag tccatgggcc aaccgcccac tccccgccag cctcgcacca
     5821 acaccaaccg gtccagtaca tgagccagtc gggtcactga ccccggcacc agtccctcag
     5881 ccactggatc cagcgcccgc agtgactccc gaggccagtc acctgttgga ggatcccgat
     5941 gaagagacga gccaggctgt caaagccctt cgggagatgg ccgatactgt gattccccag
     6001 aaggaagagg ctgcaatctg tggccaaatg gacctttccc atccgccccc aaggggccat
     6061 ctggatgagc tgacaaccac acttgagtcc atgaccgagg atctgaacct ggactcaccc
     6121 ctgaccccgg aattgaacga gattctggat accttcctga acgacgagtg cctcttgcat
     6181 gccatgcata tcagcacagg actgtccatc ttcgacacat ctctgttttg agtttaaacg
     6241 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     6301 aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc
     6361 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     6421 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     6481 ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg aaatcggatc cgcggccgca
     6541 ctgattctag ctgcagaggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag
     6601 cccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg
     6661 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag
     6721 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca
     6781 gaacacagga tcctcgagcc accatgattg aacaagatgg attgcacgca ggttctccgg
     6841 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     6901 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     6961 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     7021 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     7081 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     7141 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     7201 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     7261 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     7321 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     7381 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     7441 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     7501 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     7561 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagt taacttgttt attgcagctt
     7621 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     7681 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggaattgac
     7741 tcaaatgatg tcaattagtc tatcagaagc tcatctggtc tcccttccgg gggacaagac
     7801 atccctgttt aatatttaaa cagcagtgtt cccaaactgg gttcttatat cccttgctct
     7861 ggtcaaccag gttgcagggt ttcctgtcct cacaggaacg aagtccctaa agaaacagtg
     7921 gcagccaggt ttagccccgg aattgactgg attccttttt tagggcccat tggtatggct
     7981 ttttccccgt atccccccag gtgtctgcag gctcaaagag cagcgagaag cgttcagagg
     8041 aaagcgatcc cgtgccacct tccccgtgcc cgggctgtcc ccgcacgctg ccggctcggg
     8101 gatgcggggg gagcgccgga ccggagcgga gccccgggcg gctcgctgct gccccctagc
     8161 gggggaggga cgtaattaca tccctggggg ctttgggggg gggctgtccc tgatatctat
     8221 aacaagaaaa tatatatata ataagttatc acgtaagtag aacatgaaat aacaatataa
     8281 ttatcgtatg agttaaatct taaaagtcac gtaaaagata atcatgcgtc attttgactc
     8341 acgcggtcgt tatagttcaa aatcagtgac acttaccgca ttgacaagca cgcctcacgg
     8401 gagctccaag cggcgactga gatgtcctaa atgcacagcg acggattcgc gctatttaga
     8461 aagagagagc aatatttcaa gaatgcatgc gtcaatttta cgcagactat ctttctaggg
     8521 ttaatctagc tgcatcagga tcatatcgtc gggtcttttt tccggctcag tcatcgccca
     8581 agctggcgct atctgggcat cggggaggaa gaagcccgtg ccttttcccg cgaggttgaa
     8641 gcggcatgga aagagtttgc cgaggatgac tgctgctgca ttgacgttga gcgaaaacgc
     8701 acgtttacca tgatgattcg ggaaggtgtg gccatgcacg cctttaacgg tgaactgttc
     8761 gttcaggcca cctgggatac cagttcgtcg cggcttttcc ggacacagtt ccggatggtc
     8821 agcccgaagc gcatcagcaa cccgaacaat accggcgaca gccggaactg ccgtgccggt
     8881 gtgcagatta atgacagcgg tgcggcgctg ggatattacg tcagcgagga cgggtatcct
     8941 ggctggatgc cgcagaaatg gacatggata ccccgtgagt tacccggcgg gcgcgcttgg
     9001 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     9061 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     9121 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     9181 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     9241 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     9301 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     9361 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     9421 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     9481 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     9541 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     9601 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     9661 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     9721 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     9781 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     9841 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     9901 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     9961 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
    10021 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
    10081 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
    10141 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
    10201 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
    10261 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
    10321 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
    10381 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
    10441 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
    10501 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
    10561 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
    10621 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
    10681 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
    10741 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
    10801 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
    10861 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
    10921 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
    10981 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
    11041 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
    11101 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
    11161 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
    11221 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
    11281 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
    11341 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
    11401 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
    11461 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
    11521 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
    11581 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
    11641 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
    11701 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
    11761 taaaacgacg gccagtgagc gcgcctcgtt cattcacgtt tttgaacccg tggaggacgg
    11821 gcagactcgc ggtgcaaatg tgttttacag cgtgatggag cagatgaaga tgctcgacac
    11881 gctgcagaac acgcagctag attaacccta gaaagataat catattgtga cgtacgttaa
    11941 agataatcat gtgtaaaatt gacgcatgtg ttttatcggt ctgtatatcg aggtttattt
    12001 attaatttga atagatatta agttttatta tatttacact tacatactaa taataaattc
    12061 aacaaacaat ttatttatgt ttatttattt attaaaaaaa acaaaaactc aaaatttctt
    12121 ctataaagta acaaaacttt tatgagggac agcccccccc caaagccccc agggatgtaa
    12181 ttacgtccct cccccgctag ggggcagcag cgagccgccc ggggctccgc tccggtccgg
    12241 cgctcccccc gcatccccga gccggcagcg tgcggggaca gcccgggcac ggggaaggtg
    12301 gcacgggatc gctttcctct gaacgcttct cgctgctctt tgagcctgca gacacctggg
    12361 gggatacggg gaaaaggcct ccacggccga cattgattat tgactagtta ttaatagtaa
    12421 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
    12481 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
    12541 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
    12601 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
    12661 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
    12721 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc
    12781 cccacgttct gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta
    12841 tttatttttt aattattttg tgcagcgatg ggggcggggg gggggggggg g
//
