LOCUS       sequence-447700-        9591 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCMV and pT7 Human expression plasmid for TadCBEd using SpCas9 CBE 
            enzyme with MRKQKC amino acids at positions 
            1135,1136,1218,1219,1335,T1337 with C-term BPNLS-P2A-EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9591)
  AUTHORS   Silverstein RA, Kim N, Kroell AS, Walton RT, Delano J, Butcher RM, 
            Pacesa M, Smith BK, Christie KA, Ha LL, Meis RJ, Clark AB, Spinner 
            AD, Lazzarotto CR, Li Y, Matsubara A, Urbina EO, Dahl GA, Correia 
            BE, Marks DS, Tsai SQ, Pinello L, De Ravin SS, Liu Q, Kleinstiver BP
  TITLE     Custom CRISPR-Cas9 PAM variants via scalable engineering and machine
            learning.
  JOURNAL   Nature. 2025 Apr 22. doi: 10.1038/s41586-025-09021-y.
  PUBMED    40262634
REFERENCE   2  (bases 1 to 9591)
  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: "Nature. 
            2025 Apr 22. doi: 10.1038/s41586-025-09021-y."
COMMENT     Sequence Label: pCMV-T7-TadCBEd-SpCas9(MRKQKC)-P2A-EGFP (AHK574)
FEATURES             Location/Qualifiers
     source          1..9591
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        112..491
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        492..695
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     646..666
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     737..756
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        737..755
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             805..825
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1420..5520
                     /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
                     SILPKRNSDKLIARKKDWDPKKYGGFMRPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAKQLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKKYCSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5551..5799
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5830..6078
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6121..6141
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6151..6207
                     /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(6208..6210,6211..6213,6214..6924)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6208..6210,6211..6213,6214..6924)
                     /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(6514..6533)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6861..6882
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6937..6957
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6966..6983
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(7006..7023)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7012..7236
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7307..7323)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7307..7323)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7320..7342)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7331..7347
                     /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(7355..7361,7362..7379,7380..7385))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7400..7421
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7538..7555)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7709..8297)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7789..7808)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8468..9259,9260..9328))
                     /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     9091..9110
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9329..9433)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9512..9531)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
       61 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat
      121 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      181 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      241 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      301 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      361 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      421 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      481 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      541 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      601 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      661 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      721 agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat gaaacggaca
      781 gccgacggaa gcgagttcga gtcaccaaag aagaagcgga aagtctctga ggtggagttt
      841 tcccacgagt actggatgag acatgccctg accctggcca agagggcacg cgatgagagg
      901 aaggcccctg tgggagccgt gctggtgctg aacaatagag tgatcggcga gggctggaac
      961 agagccatcg gcctgcacga cccaacagcc catgccgaaa ttatcgccct gagacagggc
     1021 ggcctggtca tgcagaacta cagactgatt gacgccaccc tgtacgtgac attcgagcct
     1081 tgcgtgatgt gcgccggcgc catgatcaac tctaggatcg gccgcgtggt gtttggcgtg
     1141 aggaacagca aaagaggcgc cgcaggctcc ctgatgaacg tgctgaacta ccccggcatg
     1201 aatcaccgcg tcgaaattac cgagggaatc ctggcagatg aatgtgccgc cctgctgtgc
     1261 gatttctacc ggatgcctag acaggtgttc aatgctcaga agaaggccca gagctccatc
     1321 aactccggag gatctagcgg aggctcctct ggctctgaga cacctggcac aagcgagagc
     1381 gcaacacctg aaagcagcgg gggcagcagc ggggggtcag acaagaagta cagcatcggc
     1441 ctggccatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc
     1501 agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc
     1561 ggagccctgc tgttcgacag cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc
     1621 agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac
     1681 gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa
     1741 gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac
     1801 cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag
     1861 gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc
     1921 ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg
     1981 gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc
     2041 aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag
     2101 ctgcccggcg agaagaagaa tggcctgttc ggaaacctga ttgccctgag cctgggcctg
     2161 acccccaact tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag
     2221 gacacctacg acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac
     2281 ctgtttctgg ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg
     2341 aacaccgaga tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac
     2401 caccaggacc tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa
     2461 gagattttct tcgaccagag caagaacggc tacgccggct acattgacgg cggagccagc
     2521 caggaagagt tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa
     2581 ctgctcgtga agctgaacag agaggacctg ctgcggaagc agcggacctt cgacaacggc
     2641 agcatccccc accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat
     2701 ttttacccat tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc
     2761 ccctactacg tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag
     2821 agcgaggaaa ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc
     2881 cagagcttca tcgagcggat gaccaacttc gataagaacc tgcccaacga gaaggtgctg
     2941 cccaagcaca gcctgctgta cgagtacttc accgtgtata acgagctgac caaagtgaaa
     3001 tacgtgaccg agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc
     3061 gtggacctgc tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac
     3121 ttcaagaaaa tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga tcggttcaac
     3181 gcctccctgg gcacatacca cgatctgctg aaaattatca aggacaagga cttcctggac
     3241 aatgaggaaa acgaggacat tctggaagat atcgtgctga ccctgacact gtttgaggac
     3301 agagagatga tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga caaagtgatg
     3361 aagcagctga agcggcggag atacaccggc tggggcaggc tgagccggaa gctgatcaac
     3421 ggcatccggg acaagcagtc cggcaagaca atcctggatt tcctgaagtc cgacggcttc
     3481 gccaacagaa acttcatgca gctgatccac gacgacagcc tgacctttaa agaggacatc
     3541 cagaaagccc aggtgtccgg ccagggcgat agcctgcacg agcacattgc caatctggcc
     3601 ggcagccccg ccattaagaa gggcatcctg cagacagtga aggtggtgga cgagctcgtg
     3661 aaagtgatgg gccggcacaa gcccgagaac atcgtgatcg aaatggccag agagaaccag
     3721 accacccaga agggacagaa gaacagccgc gagagaatga agcggatcga agagggcatc
     3781 aaagagctgg gcagccagat cctgaaagaa caccccgtgg aaaacaccca gctgcagaac
     3841 gagaagctgt acctgtacta cctgcagaat gggcgggata tgtacgtgga ccaggaactg
     3901 gacatcaacc ggctgtccga ctacgatgtg gaccatatcg tgcctcagag ctttctgaag
     3961 gacgactcca tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg caagagcgac
     4021 aacgtgccct ccgaagaggt cgtgaagaag atgaagaact actggcggca gctgctgaac
     4081 gccaagctga ttacccagag aaagttcgac aatctgacca aggccgagag aggcggcctg
     4141 agcgaactgg ataaggccgg cttcatcaag agacagctgg tggaaacccg gcagatcaca
     4201 aagcacgtgg cacagatcct ggactcccgg atgaacacta agtacgacga gaatgacaag
     4261 ctgatccggg aagtgaaagt gatcaccctg aagtccaagc tggtgtccga tttccggaag
     4321 gatttccagt tttacaaagt gcgcgagatc aacaactacc accacgccca cgacgcctac
     4381 ctgaacgccg tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga aagcgagttc
     4441 gtgtacggcg actacaaggt gtacgacgtg cggaagatga tcgccaagag cgagcaggaa
     4501 atcggcaagg ctaccgccaa gtacttcttc tacagcaaca tcatgaactt tttcaagacc
     4561 gagattaccc tggccaacgg cgagatccgg aagcggcctc tgatcgagac aaacggcgaa
     4621 accggggaga tcgtgtggga taagggccgg gattttgcca ccgtgcggaa agtgctgagc
     4681 atgccccaag tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt cagcaaagag
     4741 tctatcctgc ccaagaggaa cagcgataag ctgatcgcca gaaagaagga ctgggaccct
     4801 aagaagtacg gcggcttcat gagacccacc gtggcctatt ctgtgctggt ggtggccaaa
     4861 gtggaaaagg gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc
     4921 atggaaagaa gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa
     4981 gaagtgaaaa aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac
     5041 ggccggaaga gaatgctggc ctctgccaag cagctgcaga agggaaacga actggccctg
     5101 ccctccaaat atgtgaactt cctgtacctg gccagccact atgagaagct gaagggctcc
     5161 cccgaggata atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag
     5221 atcatcgagc agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac
     5281 aaagtgctgt ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat
     5341 atcatccacc tgtttaccct gaccaatctg ggagcccctg ccgccttcaa gtactttgac
     5401 accaccatcg accggaagaa gtactgcagc accaaagagg tgctggacgc caccctgatc
     5461 caccagagca tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggtgac
     5521 agcggcggga gcggcgggag cggggggagc actaatctga gcgacatcat tgagaaggag
     5581 actgggaaac agctggtcat tcaggagtcc atcctgatgc tgcctgagga ggtggaggaa
     5641 gtgatcggca acaagccaga gtctgacatc ctggtgcaca ccgcctacga cgagtccaca
     5701 gatgagaatg tgatgctgct gacctctgac gcccccgagt ataagccttg ggccctggtc
     5761 atccaggatt ctaacggcga gaataagatc aagatgctga gcggaggatc cggaggatct
     5821 ggaggcagca ccaacctgtc tgacatcatc gagaaggaga caggcaagca gctggtcatc
     5881 caggagagca tcctgatgct gcccgaagaa gtcgaagaag tgatcggaaa caagcctgag
     5941 agcgatatcc tggtccatac cgcctacgac gagagtaccg acgaaaatgt gatgctgctg
     6001 acatccgacg ccccagagta taagccctgg gctctggtca tccaggattc caacggagag
     6061 aacaaaatca aaatgctgtc tggcggctca aaaagaaccg ccgacggcag cgaattcgag
     6121 cccaagaaga agaggaaagt cggaagcgga gctactaact tcagcctgct gaagcaggct
     6181 ggagacgtgg aggagaaccc tggacctatg gtgagcaagg gcgaggagct gttcaccggg
     6241 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     6301 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     6361 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctatgg agtgcagtgc
     6421 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     6481 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     6541 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     6601 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     6661 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     6721 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     6781 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
     6841 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     6901 ctcggcatgg acgagctgta caagtctggt ggttctccca agaagaagag gaaagtctaa
     6961 ccggtcatca tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt
     7021 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     7081 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     7141 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     7201 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     7261 ggggctcgat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     7321 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     7381 gtaaagccta gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     7441 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     7501 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     7561 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     7621 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     7681 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     7741 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     7801 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     7861 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     7921 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     7981 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     8041 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     8101 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     8161 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     8221 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     8281 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac actcagtgga
     8341 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     8401 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     8461 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     8521 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     8581 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     8641 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     8701 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     8761 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     8821 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     8881 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     8941 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     9001 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     9061 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     9121 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     9181 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     9241 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     9301 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     9361 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     9421 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     9481 gatctcccga tcccctaggg tcgactctca gtacaatctg ctctgatgcc gcatagttaa
     9541 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg a
//
