pCAG-CBE4max-SpCas9-NG-P2A-EGFP (RTW4554)

LOCUS       pCAG-CBE4max-SpCas9-NG-P2A-EGFP (RTW4554)       11197 bp    DNA     circular SYN 30-JUN-2025
DEFINITION  CAG promoter expression plasmid for human codon optimized BE4max 
            C-to-T base editor with 
            SpCas9-NG(D10A/L1111R/D1135V/G1218R/E1219F/A1322R/R1335V/T1337R) and
            P2A-EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11197)
  AUTHORS   Walton RT, Christie KA, Whittaker MN, Kleinstiver BP
  TITLE     Unconstrained genome targeting with near-PAMless engineered 
            CRISPR-Cas9 variants.
  JOURNAL   Science. 2020 Apr 17;368(6488):290-296. doi: 
            10.1126/science.aba8853. Epub 2020 Mar 26.
  PUBMED    32217751
REFERENCE   2  (bases 1 to 11197)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 30, 2025 from SnapGene Server 8.0.1
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1126/science.aba8853"; journalName: "Science"; date: 
            "2020-04-17- 17"; volume: "368"; issue: "6488"; pages: "290-296"
COMMENT     Sequence Label: pCAG-CBE4max-SpCas9-NG-P2A-EGFP (RTW4554)
FEATURES             Location/Qualifiers
     source          1..11197
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(3361..4044)
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="SSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGG
                     RHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLS
                     RYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNE
                     AHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHIL
                     WATGLK"
     CDS             complement(4045..4065)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4173..4192)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          4200..5216
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(4254..4276)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        5906..6010
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6011..6079,6080..6871)
                     /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     complement(6229..6248)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7042..7630
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7531..7550
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7784..7801
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    7869..8003
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7893..7912)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7947..7966
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(8009..8204)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8023..8158
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    8263..8284
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8299..8304,8305..8322,8323..8329)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8337..8353
                     /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)."
     primer_bind     8342..8364
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8361..8377
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8361..8377
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8720..8739
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    8738..8793
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     8839..8858
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(join(8945..9658,9659..9661,9662..9664))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(8948..9658,9659..9661,9662..9664))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(8990..9011)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     9339..9358
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             complement(9665..9721)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(9740..9760)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(9803..10051)
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             complement(10082..10330)
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             complement(join(10361..11197,1..3264))
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SIRPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASARFLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPRAFKYFDTTIDRKVYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
ORIGIN
        1 catgctcagc actttccgca cggtggcaaa atcccggccc ttatcccaca cgatctcccc
       61 ggtttcgccg tttgtctcga tcagaggccg cttccggatc tcgccgttgg ccagggtaat
      121 ctcggtcttg aaaaagttca tgatgttgct gtagaagaag tacttggcgg tagccttgcc
      181 gatttcctgc tcgctcttgg cgatcatctt ccgcacgtcg tacaccttgt agtcgccgta
      241 cacgaactcg ctttccagct tagggtactt tttgatcagg gcggttccca cgacggcgtt
      301 caggtaggcg tcgtgggcgt ggtggtagtt gttgatctcg cgcactttgt aaaactggaa
      361 atccttccgg aaatcggaca ccagcttgga cttcagggtg atcactttca cttcccggat
      421 cagcttgtca ttctcgtcgt acttagtgtt catccgggag tccaggatct gtgccacgtg
      481 ctttgtgatc tgccgggttt ccaccagctg tctcttgatg aagccggcct tatccagttc
      541 gctcaggccg cctctctcgg ccttggtcag attgtcgaac tttctctggg taatcagctt
      601 ggcgttcagc agctgccgcc agtagttctt catcttcttc acgacctctt cggagggcac
      661 gttgtcgctc ttgccccggt tcttgtcgct tctggtcagc accttgttgt cgatggagtc
      721 gtccttcaga aagctctgag gcacgatatg gtccacatcg tagtcggaca gccggttgat
      781 gtccagttcc tggtccacgt acatatcccg cccattctgc aggtagtaca ggtacagctt
      841 ctcgttctgc agctgggtgt tttccacggg gtgttctttc aggatctggc tgcccagctc
      901 tttgatgccc tcttcgatcc gcttcattct ctcgcggctg ttcttctgtc ccttctgggt
      961 ggtctggttc tctctggcca tttcgatcac gatgttctcg ggcttgtgcc ggcccatcac
     1021 tttcacgagc tcgtccacca ccttcactgt ctgcaggatg cccttcttaa tggcggggct
     1081 gccggccaga ttggcaatgt gctcgtgcag gctatcgccc tggccggaca cctgggcttt
     1141 ctggatgtcc tctttaaagg tcaggctgtc gtcgtggatc agctgcatga agtttctgtt
     1201 ggcgaagccg tcggacttca ggaaatccag gattgtcttg ccggactgct tgtcccggat
     1261 gccgttgatc agcttccggc tcagcctgcc ccagccggtg tatctccgcc gcttcagctg
     1321 cttcatcact ttgtcgtcga acaggtgggc ataggttttc agccgttcct cgatcatctc
     1381 tctgtcctca aacagtgtca gggtcagcac gatatcttcc agaatgtcct cgttttcctc
     1441 attgtccagg aagtccttgt ccttgataat tttcagcaga tcgtggtatg tgcccaggga
     1501 ggcgttgaac cgatcttcca cgccggagat ttccacggag tcgaagcact cgattttctt
     1561 gaagtagtcc tctttcagct gcttcacggt cactttccgg ttggtcttga acagcaggtc
     1621 cacgatggcc tttttctgct cgccgctcag gaaggcgggc tttctcattc cctcggtcac
     1681 gtatttcact ttggtcagct cgttatacac ggtgaagtac tcgtacagca ggctgtgctt
     1741 gggcagcacc ttctcgttgg gcaggttctt atcgaagttg gtcatccgct cgatgaagct
     1801 ctgggcggaa gcgcccttgt ccaccacttc ctcgaagttc cagggggtga tggtttcctc
     1861 gctctttctg gtcatccagg cgaatctgct gtttcccctg gccagagggc ccacgtagta
     1921 ggggatgcgg aaggtcagga tcttctcgat cttttcccgg ttgtccttca ggaatgggta
     1981 aaaatcttcc tgccgccgca gaatggcgtg cagctctccc aggtggatct ggtgggggat
     2041 gctgccgttg tcgaaggtcc gctgcttccg cagcaggtcc tctctgttca gcttcacgag
     2101 cagttcctcg gtgccgtcca tcttttccag gatgggcttg atgaacttgt agaactcttc
     2161 ctggctggct ccgccgtcaa tgtagccggc gtagccgttc ttgctctggt cgaagaaaat
     2221 ctctttgtac ttctcaggca gctgctgccg cacgagagct ttcagcaggg tcaggtcctg
     2281 gtggtgctcg tcgtatctct tgatcataga ggcgctcagg ggggccttgg tgatctcggt
     2341 gttcactctc aggatgtcgc tcagcaggat ggcgtcggac aggttcttgg cggccagaaa
     2401 caggtcggcg tactggtcgc cgatctgggc cagcaggttg tccaggtcgt cgtcgtaggt
     2461 gtccttgctc agctgcagtt tggcatcctc ggccaggtcg aagttgctct tgaagttggg
     2521 ggtcaggccc aggctcaggg caatcaggtt tccgaacagg ccattcttct tctcgccggg
     2581 cagctgggcg atcagatttt ccagccgtct gctcttgctc agtctggcag acaggatggc
     2641 cttggcgtcc acgccgctgg cgttgatggg gttttcctcg aacagctggt tgtaggtctg
     2701 caccagctgg atgaacagct tgtccacgtc gctgttgtcg gggttcaggt cgccctcgat
     2761 caggaagtgg ccccggaact tgatcatgtg ggccagggcc agatagatca gccgcaggtc
     2821 ggccttgtcg gtgctgtcca ccagtttctt tctcaggtgg tagatggtgg ggtacttctc
     2881 gtggtaggcc acctcgtcca cgatgttgcc gaagatgggg tgccgctcgt gcttcttatc
     2941 ctcttccacc aggaaggact cttccagtct gtggaagaag ctgtcgtcca ccttggccat
     3001 ctcgttgctg aagatctctt gcagatagca gatccggttc ttccgtctgg tgtatcttct
     3061 tctggcggtt ctcttcagcc gggtggcctc ggctgtttcg ccgctgtcga acagcagggc
     3121 tccgatcagg ttcttcttga tgctgtgccg gtcggtgttg cccagcacct tgaatttctt
     3181 gctgggcacc ttgtactcgt cggtgatcac ggcccagccc acagagttgg tgccgatggc
     3241 caggccgatg ctgtacttct tgtcgctgcc gccgctgctg ccgccactgc tctctggtgt
     3301 tgctgactcg cttgttcctg gtgtctcgct gccagaggat cctccgctag atcctccaga
     3361 cttcaggcct gtggcccaca ggatgtgggg tggcagcctc tgatagtgac aagactgcag
     3421 ggcgattgta aagaaggtca gctggggctg ctttctccgc aggatattca gacaaggggg
     3481 caggcccagg atgatgcaat acagctccag cacgtacagg cgcacccaca ggtgtgggta
     3541 cctaggccag tgggcctcgt tgctaggaga ataattcacg aagttccgcc agcagtagcc
     3601 ggactcctgc tctgtcatga tctggatggt cacgccggag ctgatcagat cgcgcaggcc
     3661 ctgcctattc cttgggtctg cgtggtgata cagcctggcg atgtagataa acagtgtcac
     3721 gtgtggatat ctggacagga actcggtgat ggccctgcta cactctccgc aaggggacca
     3781 gctcagaaac catgtgatgc tacatctggt attggggcag aagtaccgct ctgtggtaaa
     3841 cttctcgatg aagttcacct ccacgtgctt atttgtgttc tgagaggtgt gcctccagat
     3901 ggagtgcctg cctccccagt tgatctcgta cagcaggcat gtctcctttc tcagctcccg
     3961 ggggtcaaag aacacttcaa actcgtgagg ttcaatccgg cggcgcaggg ttggatcgac
     4021 ggcgacaggc ccagtctctg aggagacttt ccgcttcttc tttggtgact cgaactcgct
     4081 tccgtcggct gtccgtttca tggtggcgac cggtggatcc cgggcccgcg gtaccgtcga
     4141 ctgcagaatt ctttgccaaa atgatgagac agcacaataa ccagcacgtt gcccaggagc
     4201 tgtaggaaaa agaagaaggc atgaacatgg ttagcagagg ctctagagcc gccggtcaca
     4261 cgccagaagc cgaaccccgc cctgccccgt cccccccgaa ggcagccgtc cccccgcgga
     4321 cagccccgag gctggagagg gagaagggga cggcggcgcg gcgacgcacg aaggccctcc
     4381 ccgcccattt ccttcctgcc ggcgccgcac cgcttcgccc cgcgcccgct agagggggtg
     4441 cggcggcgcc tcccagattt cggctccgca cagatttggg acaaaggaag tccctgcgcc
     4501 ctctcgcacg attaccataa aaggcaatgg ctgcggctcg ccgcgcctcg acagccgccg
     4561 gcgctccggg ggccgccgcg cccctccccc gagccctccc cggcccgagg cggccccgcc
     4621 ccgcccggca cccccacctg ccgccacccc ccgcccggca cggcgagccc cgcgccacgc
     4681 cccgtacgga gccccgcacc cgaagccggg ccgtgctcag caactcgggg aggggggtgc
     4741 agggggggtt gcagcccgac cgacgcgccc acaccccctg ctcacccccc cacgcacaca
     4801 ccccgcacgc agcctttgtt cccctcgcag ccccccccgc accgcggggc accgcccccg
     4861 gccgcgctcc cctcgcgcac actgcggagc gcacaaagcc ccgcgccgcg cccgcagcgc
     4921 tcacagccgc cgggcagcgc ggagccgcac gcggcgctcc ccacgcacac acacacgcac
     4981 gcaccccccg agccgctccc cccgcacaaa gggccctccc ggagcccctc aaggctttca
     5041 cgcagccaca gaaaagaaac aagccgtcat taaaccaagc gctaattaca gcccggagga
     5101 gaagggccgt cccgcccgct cacctgtggg agtaacgcgg tcagtcagag ccggggcggg
     5161 cggcgcgagg cggcggcgga gcggggcacg gggcgaaggc agcgcgcagc gactcccgcc
     5221 cgccgcgcgc ttcgcttttt atagggccgc cgccgccgcc gcctcgccat aaaaggaaac
     5281 tttcggagcg cgccgctctg attggctgcc gccgcacctc tccgcctcgc cccgccccgc
     5341 ccctcgcccc gccccgcccc gcctggcgcg cgcccccccc cccccccccc cccccccccc
     5401 ccccaaaaaa ataaaaaaaa aaaaaaaaaa aaaaatgggg gggggggggg gggggggggg
     5461 ggggggggga gggggggggg gggggggggc cgcgccgggg ggaatagggg caccaaaaac
     5521 tacttggtac accaagagga aaaggcccaa aaggccattt catgggccta atgcaacgcg
     5581 gggccttttc agtccttatc gtcaataggg ggcgtacttg gcatatgata cacttgatgt
     5641 actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct
     5701 attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc
     5761 ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta
     5821 tgaactaatg accccgtaat tgattactat taataactag tcaataatca atgtcgaccc
     5881 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     5941 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     6001 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     6061 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     6121 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     6181 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     6241 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     6301 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     6361 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     6421 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     6481 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     6541 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     6601 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     6661 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     6721 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     6781 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     6841 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     6901 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6961 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     7021 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     7081 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     7141 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     7201 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     7261 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     7321 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     7381 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     7441 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     7501 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     7561 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     7621 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     7681 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     7741 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     7801 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     7861 atgcagcgga tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     7921 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     7981 ttataagctg caataaacaa gttagctttt tgcaaaagcc taggcctcca aaaaagcctc
     8041 ctcactactt ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa
     8101 aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg
     8161 gcggagttag gggcgggact atggttgctg actaattgag atgcggatcc gctggcacga
     8221 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     8281 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     8341 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcttgggct
     8401 gcaggtcgag ggatctccat aagagaagag ggacagctat gactgggagt agtcaggaga
     8461 ggaggaaaaa tctggctagt aaaacatgta aggaaaattt tagggatgtt aaagaaaaaa
     8521 ataacacaaa acaaaatata aaaaaaatct aacctcaagt caaggctttt ctatggaata
     8581 aggaatggac agcagggggc tgtttcatat actgatgacc tctttatagc caacctttgt
     8641 tcatggcagc cagcatatgg gcatatgttg ccaaactcta aaccaaatac tcattctgat
     8701 gttttaaatg atttgccctc ccatatgtcc ttccgagtga gagacacaaa aaattccaac
     8761 acactattgc aatgaaaata aatttccttt attagccaga agtcagatgc tcaaggggct
     8821 tcatgatgtc cccataattt ttggcagagg gaaaaagatc tcagtggtat ttgtgagcca
     8881 gggcattggc cacaccagcc accaccttct gataggcagc ctgcacctga ggagtgcggc
     8941 cgcttcactt gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca
     9001 gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct cagggcggac tgggtgctca
     9061 ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt
     9121 ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag ttcaccttga
     9181 tgccgttctt ctgcttgtcg gccatgatat agacgttgtg gctgttgtag ttgtactcca
     9241 gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt
     9301 tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct
     9361 tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg aagaagtcgt
     9421 gctgcttcat gtggtcgggg tagcggctga agcactgcac gccgtaggtc agggtggtca
     9481 cgagggtggg ccagggcacg ggcagcttgc cggtggtgca gatgaacttc agggtcagct
     9541 tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt
     9601 cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg cccttgctca
     9661 ccatcggtcc aggattctct tcgacatctc cggcttgttt cagcagagag aagtttgttg
     9721 cgccgcttcc accgcctccg actttcctct tcttcttggg ctcgaattcg ctgccgtcgg
     9781 cggttctttt tgagccgcca gacagcattt tgattttgtt ctctccgttg gaatcctgga
     9841 tgaccagagc ccagggctta tactctgggg cgtcggatgt cagcagcatc acattttcgt
     9901 cggtactctc gtcgtaggcg gtatggacca ggatatcgct ctcaggcttg tttccgatca
     9961 cttcttcgac ttcttcgggc agcatcagga tgctctcctg gatgaccagc tgcttgcctg
    10021 tctccttctc gatgatgtca gacaggttgg tgctgcctcc agatcctccg gatcctccgc
    10081 tcagcatctt gatcttattc tcgccgttag aatcctggat gaccagggcc caaggcttat
    10141 actcgggggc gtcagaggtc agcagcatca cattctcatc tgtggactcg tcgtaggcgg
    10201 tgtgcaccag gatgtcagac tctggcttgt tgccgatcac ttcctccacc tcctcaggca
    10261 gcatcaggat ggactcctga atgaccagct gtttcccagt ctccttctca atgatgtcgc
    10321 tcagattagt gctccccccg ctcccgccgc tcccgccgct gtcacctccc agctgagaca
    10381 ggtcgatccg tgtctcgtac aggccggtga tgctctggtg gatcagggtg gcgtccagca
    10441 cctctttggt gcttctgtac accttccggt cgatggtggt gtcaaagtac ttgaaggctc
    10501 taggggctcc cagattggtc agggtaaaca ggtggatgat attctcggcc tgctctctga
    10561 tgggcttatc ccggtgcttg ttgtaggcgg acagcacttt gtccagatta gcgtcggcca
    10621 ggatcactct cttggagaac tcgctgatct gctcgatgat ctcgtccagg tagtgcttgt
    10681 gctgttccac aaacagctgt ttctgctcat tatcctcggg ggagcccttc agcttctcat
    10741 agtggctggc caggtacagg aagttcacat atttggaggg cagggccagt tcgtttccct
    10801 tctgcaggaa tctggcagag gccagcattc tcttccggcc gttttccagc tcgaacaggg
    10861 agtacttagg cagcttgatg atcaggtcct ttttcacttc tttgtagccc ttggcttcca
    10921 gaaagtcgat gggattcttc tcgaagctgc ttctttccat gatggtgatc cccagcagct
    10981 ctttcacact cttcagtttc ttggacttgc ccttttccac tttggccacc accagcacag
    11041 aataggccac ggtggggctc acgaagccgc cgtacttctt agggtcccag tccttctttc
    11101 tggcgatcag cttatcgctg ttcctcttgg gtctgataga ctctttgctg aagccgcctg
    11161 tctgcacctc ggtctttttc acgatattca cttgggg
//