pCAG-CBE4max-SpG-P2A-EGFP (RTW4552)

LOCUS       pCAG-CBE4max-SpG-P2A-EGFP (RTW4552)               11195 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  CAG promoter expression plasmid for human codon optimized BE4max 
            C-to-T base editor with 
            SpG(D10A/D1135L/S1136W/G1218K/E1219Q/R1335Q/T1337R) and P2A-EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    pCAG-CBE4max-SpG-P2A-EGFP (RTW4552)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11195)
  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 11195)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..11195
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     intron          161..1177
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1101..1123
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1185..1204
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1312..1332
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1333..2016
                     /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             2113..3828
                     /codon_start=1
                     /product="N-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(N)
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
     CDS             6244..6492
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6523..6771
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6814..6834
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6853..6909
                     /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             6910..7626
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6955..6976)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7216..7235)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7563..7584
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(7716..7735)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7781..7836
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7835..7854)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(8197..8213)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8197..8213)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8210..8232)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8221..8237
                     /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(8245..8275)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8290..8311
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8370..8565
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      8416..8551
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8478..8497
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    8571..8705
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8608..8627)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8662..8681
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8773..8790)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8944..9532)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9024..9043)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(9703..10563)
                     /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     10326..10345
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10564..10668)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        10699..11078
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        11080..160
                     /label=chicken beta-actin promoter
ORIGIN
        1 ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg ggcggggcga ggcggagagg
       61 tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg
      121 gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct
      181 tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc
      241 gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt
      301 ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa agccttgagg ggctccggga
      361 gggccctttg tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag
      421 cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct
      481 ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc cgcggtgcgg
      541 ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg ggtgagcagg
      601 gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc gagttgctga
      661 gcacggcccg gcttcgggtg cggggctccg tacggggcgt ggcgcggggc tcgccgtgcc
      721 gggcgggggg tggcggcagg tgggggtgcc gggcggggcg gggccgcctc gggccgggga
      781 gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag
      841 ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc ctttgtccca
      901 aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc
      961 gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg
     1021 ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg gctgccttcg
     1081 ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc
     1141 tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat
     1201 tgtgctgtct catcattttg gcaaagaatt ctgcagtcga cggtaccgcg ggcccgggat
     1261 ccaccggtcg ccaccatgaa acggacagcc gacggaagcg agttcgagtc accaaagaag
     1321 aagcggaaag tctcctcaga gactgggcct gtcgccgtcg atccaaccct gcgccgccgg
     1381 attgaacctc acgagtttga agtgttcttt gacccccggg agctgagaaa ggagacatgc
     1441 ctgctgtacg agatcaactg gggaggcagg cactccatct ggaggcacac ctctcagaac
     1501 acaaataagc acgtggaggt gaacttcatc gagaagttta ccacagagcg gtacttctgc
     1561 cccaatacca gatgtagcat cacatggttt ctgagctggt ccccttgcgg agagtgtagc
     1621 agggccatca ccgagttcct gtccagatat ccacacgtga cactgtttat ctacatcgcc
     1681 aggctgtatc accacgcaga cccaaggaat aggcagggcc tgcgcgatct gatcagctcc
     1741 ggcgtgacca tccagatcat gacagagcag gagtccggct actgctggcg gaacttcgtg
     1801 aattattctc ctagcaacga ggcccactgg cctaggtacc cacacctgtg ggtgcgcctg
     1861 tacgtgctgg agctgtattg catcatcctg ggcctgcccc cttgtctgaa tatcctgcgg
     1921 agaaagcagc cccagctgac cttctttaca atcgccctgc agtcttgtca ctatcagagg
     1981 ctgccacccc acatcctgtg ggccacaggc ctgaagtctg gaggatctag cggaggatcc
     2041 tctggcagcg agacaccagg aacaagcgag tcagcaacac cagagagcag tggcggcagc
     2101 agcggcggca gcgacaagaa gtacagcatc ggcctggcca tcggcaccaa ctctgtgggc
     2161 tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac
     2221 accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga cagcggcgaa
     2281 acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac
     2341 cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc
     2401 ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc
     2461 atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac
     2521 ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc
     2581 ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac
     2641 aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag
     2701 gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc
     2761 aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg
     2821 ttcggaaacc tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac
     2881 ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac
     2941 ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc
     3001 gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg
     3061 agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct
     3121 ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac
     3181 ggctacgccg gctacattga cggcggagcc agccaggaag agttctacaa gttcatcaag
     3241 cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac
     3301 ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga
     3361 gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg
     3421 gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg
     3481 ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac
     3541 ttcgaggaag tggtggacaa gggcgcttcc gcccagagct tcatcgagcg gatgaccaac
     3601 ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac
     3661 ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc
     3721 gccttcctga gcggcgagca gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg
     3781 aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc
     3841 gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg
     3901 ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa
     3961 gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa
     4021 acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc
     4081 ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag
     4141 acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc
     4201 cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc
     4261 gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc
     4321 ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag
     4381 aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc
     4441 cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa
     4501 gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag
     4561 aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat
     4621 gtggaccata tcgtgcctca gagctttctg aaggacgact ccatcgacaa caaggtgctg
     4681 accagaagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag
     4741 aagatgaaga actactggcg gcagctgctg aacgccaagc tgattaccca gagaaagttc
     4801 gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc
     4861 aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc
     4921 cggatgaaca ctaagtacga cgagaatgac aagctgatcc gggaagtgaa agtgatcacc
     4981 ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag
     5041 atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg
     5101 atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac
     5161 gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc
     5221 ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc
     5281 cggaagcggc ctctgatcga gacaaacggc gaaaccgggg agatcgtgtg ggataagggc
     5341 cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag
     5401 accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgat
     5461 aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt cctgtggccc
     5521 accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg
     5581 aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat
     5641 cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag
     5701 ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc
     5761 aagcagctgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac
     5821 ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg
     5881 tttgtggaac agcacaagca ctacctggac gagatcatcg agcagatcag cgagttctcc
     5941 aagagagtga tcctggccga cgctaatctg gacaaagtgc tgtccgccta caacaagcac
     6001 cgggataagc ccatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat
     6061 ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa gcagtacaga
     6121 agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag
     6181 acacggatcg acctgtctca gctgggaggt gacagcggcg ggagcggcgg gagcgggggg
     6241 agcactaatc tgagcgacat cattgagaag gagactggga aacagctggt cattcaggag
     6301 tccatcctga tgctgcctga ggaggtggag gaagtgatcg gcaacaagcc agagtctgac
     6361 atcctggtgc acaccgccta cgacgagtcc acagatgaga atgtgatgct gctgacctct
     6421 gacgcccccg agtataagcc ttgggccctg gtcatccagg attctaacgg cgagaataag
     6481 atcaagatgc tgagcggagg atccggagga tctggaggca gcaccaacct gtctgacatc
     6541 atcgagaagg agacaggcaa gcagctggtc atccaggaga gcatcctgat gctgcccgaa
     6601 gaagtcgaag aagtgatcgg aaacaagcct gagagcgata tcctggtcca taccgcctac
     6661 gacgagagta ccgacgaaaa tgtgatgctg ctgacatccg acgccccaga gtataagccc
     6721 tgggctctgg tcatccagga ttccaacgga gagaacaaaa tcaaaatgct gtctggcggc
     6781 tcaaaaagaa ccgccgacgg cagcgaattc gagcccaaga agaagaggaa agtcggaggc
     6841 ggtggaagcg gcgcaacaaa cttctctctg ctgaaacaag ccggagatgt cgaagagaat
     6901 cctggaccga tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     6961 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     7021 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     7081 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     7141 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     7201 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     7261 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     7321 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     7381 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     7441 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     7501 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     7561 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     7621 tacaagtgaa gcggccgcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt
     7681 gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat
     7741 ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc
     7801 attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca
     7861 aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg
     7921 ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct
     7981 gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt
     8041 tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc
     8101 agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga
     8161 tccctcgacc tgcagcccaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     8221 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     8281 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     8341 gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt agtcagcaac catagtcccg
     8401 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     8461 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     8521 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct aacttgttta
     8581 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     8641 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     8701 ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     8761 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     8821 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     8881 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     8941 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     9001 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     9061 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     9121 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     9181 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     9241 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     9301 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     9361 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     9421 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     9481 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     9541 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     9601 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     9661 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     9721 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     9781 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     9841 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     9901 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     9961 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
    10021 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
    10081 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
    10141 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
    10201 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
    10261 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
    10321 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
    10381 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
    10441 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
    10501 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
    10561 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
    10621 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
    10681 aaaagtgcca cctgggtcga cattgattat tgactagtta ttaatagtaa tcaattacgg
    10741 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
    10801 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
    10861 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
    10921 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
    10981 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
    11041 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct
    11101 gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt
    11161 aattattttg tgcagcgatg ggggcggggg ggggg
//