LOCUS       sequence-459010-       10326 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR CBE plasmid (C to T edits): eA3A-BE5-NG engineered to include
            the sgRNA cloning backbone from PX458 and EGFP. Includes BbsI sites 
            for sgRNA cloning downstream of U6 promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10326)
  AUTHORS   Dickson KA, Field N, Blackman T, Ma Y, Xie T, Kurangil E, Idrees S, 
            Rathnayake SNH, Mahbub RM, Faiz A, Marsh DJ
  TITLE     CRISPR single base-editing: in silico predictions to variant clonal 
            cell lines.
  JOURNAL   Hum Mol Genet. 2023 Jun 27:ddad105. doi: 10.1093/hmg/ddad105.
  PUBMED    37369005
REFERENCE   2  (bases 1 to 10326)
  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: "Hum Mol 
            Genet. 2023 Jun 27:ddad105. doi: 10.1093/hmg/ddad105."
COMMENT     Sequence Label: pCMV-eA3A-BE5-(NG)-PX458-EGFP
FEATURES             Location/Qualifiers
     source          1..10326
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        63..366
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        367..570
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     521..541
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     612..631
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        612..630
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             680..700
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1391..5491
                     /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"
     CDS             5522..5770
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5801..6049
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6092..6112
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6122..6178
                     /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             join(6179..6181,6182..6184,6185..6895)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6179..6181,6182..6184,6185..6895)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6485..6504)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6832..6853
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6908..6928
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6965..6982)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6971..7178
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   7187..7327
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7187..7316
                     /label=AAV2 ITR
     rep_origin      7402..7857
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7489..7508)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7699..7720
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7874..7893)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7993..8015
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8053..8071)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8139..8243
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8244..8312,8313..9104)
                     /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(8462..8481)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9275..9863
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9764..9783
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        9925..10173
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     9925..9945
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     10096..10115
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        10192..10267
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
       61 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      121 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      181 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      241 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      301 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      361 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      421 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      481 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      541 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagaga
      601 tccgcggccg ctaatacgac tcactatagg gagagccgcc accatgaaac ggacagccga
      661 cggaagcgag ttcgagtcac caaagaagaa gcggaaagtc gaggcaagcc cagcaagcgg
      721 cccacggcac ctgatggacc cccacatttt caccagcaac ttcaacaacg gaatcggacg
      781 acacaagacc tacctgtgct atgaggtgga gaggctggac aacggcgacg ccgtgaagat
      841 ggatcagcac agaggcttcc tgcacggcca ggccaagaat ctgctgtgcg gcttctacgg
      901 ccggcacgca gagctgcgct ttctggacct ggtgccatct ctgcagctgg atcccgccca
      961 gatctatagg gtgacctggt tcatcagctg gtccccttgc tttagctggg gatgtgcagg
     1021 agaggtgagg gccttcctgc aggagaacac acacgtgagg ctgagaatct ttgccgcccg
     1081 gatctacgac tatgatcccc tgtacaagga ggccctgcag atgctgaggg acgcaggagc
     1141 ccaggtgtct atcatgacct atgatgagtt caagcactgc tgggacacat ttgtggatca
     1201 ccagggctgt ccctttcagc cttgggacgg cctggacgag cattcccagg cactgagcgg
     1261 acggctgagg gcaattctgc agaaccaggg caattctggc ggatctagcg gaggatcctc
     1321 tggcagcgag acaccaggaa caagcgagtc agcaacacca gagagcagtg gcggcagcag
     1381 cggcggcagc gacaagaagt acagcatcgg cctggccatc ggcaccaact ctgtgggctg
     1441 ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac
     1501 cgaccggcac agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac
     1561 agccgaggcc acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg
     1621 gatctgctat ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt
     1681 ccacagactg gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat
     1741 cttcggcaac atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct
     1801 gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct
     1861 ggcccacatg atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa
     1921 cagcgacgtg gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga
     1981 aaaccccatc aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa
     2041 gagcagacgg ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt
     2101 cggaaacctg attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct
     2161 ggccgaggat gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct
     2221 gctggcccag atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga
     2281 cgccatcctg ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag
     2341 cgcctctatg atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct
     2401 cgtgcggcag cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg
     2461 ctacgccggc tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc
     2521 catcctggaa aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct
     2581 gctgcggaag cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga
     2641 gctgcacgcc attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga
     2701 aaagatcgag aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg
     2761 aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt
     2821 cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt
     2881 cgataagaac ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt
     2941 caccgtgtat aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc
     3001 cttcctgagc ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa
     3061 agtgaccgtg aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt
     3121 ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct
     3181 gaaaattatc aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga
     3241 tatcgtgctg accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac
     3301 ctatgcccac ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg
     3361 ctggggcagg ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac
     3421 aatcctggat ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca
     3481 cgacgacagc ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga
     3541 tagcctgcac gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct
     3601 gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa
     3661 catcgtgatc gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg
     3721 cgagagaatg aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga
     3781 acaccccgtg gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa
     3841 tgggcgggat atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt
     3901 ggaccatatc gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac
     3961 cagaagcgac aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa
     4021 gatgaagaac tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga
     4081 caatctgacc aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa
     4141 gagacagctg gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg
     4201 gatgaacact aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct
     4261 gaagtccaag ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat
     4321 caacaactac caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat
     4381 caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt
     4441 gcggaagatg atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt
     4501 ctacagcaac atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg
     4561 gaagcggcct ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg
     4621 ggattttgcc accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac
     4681 cgaggtgcag acaggcggct tcagcaaaga gtctatccgg cccaagagga acagcgataa
     4741 gctgatcgcc agaaagaagg actgggaccc taagaagtac ggcggcttcg tcagccccac
     4801 cgtggcctat tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa
     4861 gagtgtgaaa gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc
     4921 catcgacttt ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct
     4981 gcctaagtac tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgcccg
     5041 cttcctgcag aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct
     5101 ggccagccac tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt
     5161 tgtggaacag cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa
     5221 gagagtgatc ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg
     5281 ggataagccc atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct
     5341 gggagcccct cgcgccttca agtactttga caccaccatc gaccggaagg tgtaccgcag
     5401 caccaaagag gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac
     5461 acggatcgac ctgtctcagc tgggaggtga cagcggcggg agcggcggga gcggggggag
     5521 cactaatctg agcgacatca ttgagaagga gactgggaaa cagctggtca ttcaggagtc
     5581 catcctgatg ctgcctgagg aggtggagga agtgatcggc aacaagccag agtctgacat
     5641 cctggtgcac accgcctacg acgagtccac agatgagaat gtgatgctgc tgacctctga
     5701 cgcccccgag tataagcctt gggccctggt catccaggat tctaacggcg agaataagat
     5761 caagatgctg agcggaggat ccggaggatc tggaggcagc accaacctgt ctgacatcat
     5821 cgagaaggag acaggcaagc agctggtcat ccaggagagc atcctgatgc tgcccgaaga
     5881 agtcgaagaa gtgatcggaa acaagcctga gagcgatatc ctggtccata ccgcctacga
     5941 cgagagtacc gacgaaaatg tgatgctgct gacatccgac gccccagagt ataagccctg
     6001 ggctctggtc atccaggatt ccaacggaga gaacaaaatc aaaatgctgt ctggcggctc
     6061 aaaaagaacc gccgacggca gcgaattcga gcccaagaag aagaggaaag tcggaagcgg
     6121 agctactaac ttcagcctgc tgaagcaggc tggagacgtg gaggagaacc ctggacctat
     6181 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     6241 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     6301 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     6361 cgtgaccacc ctgacctatg gagtgcagtg cttcagccgc taccccgacc acatgaagca
     6421 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     6481 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     6541 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     6601 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     6661 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     6721 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     6781 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     6841 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtctgg
     6901 tggttctccc aagaagaaga ggaaagtcta accggtggaa ttctaactag agctcgctga
     6961 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     7021 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     7081 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     7141 ggggaggatt gggaagagaa tagcaggcat gctggggagc ggccgcagga acccctagtg
     7201 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     7261 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     7321 ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     7381 atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     7441 tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt
     7501 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     7561 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg
     7621 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     7681 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct
     7741 cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg
     7801 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat
     7861 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     7921 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     7981 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     8041 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     8101 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     8161 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     8221 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     8281 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     8341 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     8401 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     8461 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     8521 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     8581 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     8641 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     8701 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     8761 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     8821 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     8881 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     8941 ctggagccgg tgagcgtgga agccgcggta tcattgcagc actggggcca gatggtaagc
     9001 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     9061 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     9121 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     9181 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     9241 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     9301 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     9361 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     9421 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     9481 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     9541 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     9601 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     9661 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     9721 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     9781 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     9841 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     9901 tttgctggcc ttttgctcac atgtgagggc ctatttccca tgattccttc atatttgcat
     9961 atacgataca aggctgttag agagataatt ggaattaatt tgactgtaaa cacaaagata
    10021 ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc agttttaaaa
    10081 ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt cgatttcttg
    10141 gctttatata tcttgtggaa aggacgaaac accgggtctt cgagaagacc tgttttagag
    10201 ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag
    10261 tcggtgcttt tttgttttag agctagaaat agcaagttaa aataaggcta gtccgttttt
    10321 agcgcg
//
