LOCUS       sequence-451166-        9591 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCMV and pT7 Human expression plasmid for TadCBEd using SpCas9 CBE 
            enzyme with ARGIMR 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(ARGIMR)-P2A-EGFP (BKS1036)
FEATURES             Location/Qualifiers
     source          1..9591
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        102..481
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        482..685
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     636..656
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     727..746
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        727..745
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             795..815
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1410..5510
                     /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
                     SILPKRNSDKLIARKKDWDPKKYGGFARPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGILQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKMYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5541..5789
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5820..6068
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6111..6131
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6141..6197
                     /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(6198..6200,6201..6203,6204..6914)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6198..6200,6201..6203,6204..6914)
                     /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(6504..6523)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6851..6872
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             6927..6947
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6956..6973
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6996..7013)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7002..7226
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7297..7313)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7297..7313)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7310..7332)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7321..7337
                     /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(7345..7351,7352..7369,7370..7375))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7390..7411
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7528..7545)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7699..8287)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7779..7798)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8458..9249,9250..9318))
                     /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     9081..9100
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9319..9423)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9502..9521)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
       61 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag
      121 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      181 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      241 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      301 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      361 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      421 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      481 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
      541 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      601 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
      661 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agagatccgc
      721 ggccgctaat acgactcact atagggagag ccgccaccat gaaacggaca gccgacggaa
      781 gcgagttcga gtcaccaaag aagaagcgga aagtctctga ggtggagttt tcccacgagt
      841 actggatgag acatgccctg accctggcca agagggcacg cgatgagagg aaggcccctg
      901 tgggagccgt gctggtgctg aacaatagag tgatcggcga gggctggaac agagccatcg
      961 gcctgcacga cccaacagcc catgccgaaa ttatcgccct gagacagggc ggcctggtca
     1021 tgcagaacta cagactgatt gacgccaccc tgtacgtgac attcgagcct tgcgtgatgt
     1081 gcgccggcgc catgatcaac tctaggatcg gccgcgtggt gtttggcgtg aggaacagca
     1141 aaagaggcgc cgcaggctcc ctgatgaacg tgctgaacta ccccggcatg aatcaccgcg
     1201 tcgaaattac cgagggaatc ctggcagatg aatgtgccgc cctgctgtgc gatttctacc
     1261 ggatgcctag acaggtgttc aatgctcaga agaaggccca gagctccatc aactccggag
     1321 gatctagcgg aggctcctct ggctctgaga cacctggcac aagcgagagc gcaacacctg
     1381 aaagcagcgg gggcagcagc ggggggtcag acaagaagta cagcatcggc ctggccatcg
     1441 gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc agcaagaaat
     1501 tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc ggagccctgc
     1561 tgttcgacag cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc agaagaagat
     1621 acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac gagatggcca
     1681 aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa gaggataaga
     1741 agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt
     1801 accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag gccgacctgc
     1861 ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc ctgatcgagg
     1921 gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg gtgcagacct
     1981 acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc aaggccatcc
     2041 tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag ctgcccggcg
     2101 agaagaagaa tggcctgttc ggaaacctga ttgccctgag cctgggcctg acccccaact
     2161 tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag gacacctacg
     2221 acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac ctgtttctgg
     2281 ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg aacaccgaga
     2341 tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac caccaggacc
     2401 tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa gagattttct
     2461 tcgaccagag caagaacggc tacgccggct acattgacgg cggagccagc caggaagagt
     2521 tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa ctgctcgtga
     2581 agctgaacag agaggacctg ctgcggaagc agcggacctt cgacaacggc agcatccccc
     2641 accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat ttttacccat
     2701 tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc ccctactacg
     2761 tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag agcgaggaaa
     2821 ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc cagagcttca
     2881 tcgagcggat gaccaacttc gataagaacc tgcccaacga gaaggtgctg cccaagcaca
     2941 gcctgctgta cgagtacttc accgtgtata acgagctgac caaagtgaaa tacgtgaccg
     3001 agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc
     3061 tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac ttcaagaaaa
     3121 tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga tcggttcaac gcctccctgg
     3181 gcacatacca cgatctgctg aaaattatca aggacaagga cttcctggac aatgaggaaa
     3241 acgaggacat tctggaagat atcgtgctga ccctgacact gtttgaggac agagagatga
     3301 tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga caaagtgatg aagcagctga
     3361 agcggcggag atacaccggc tggggcaggc tgagccggaa gctgatcaac ggcatccggg
     3421 acaagcagtc cggcaagaca atcctggatt tcctgaagtc cgacggcttc gccaacagaa
     3481 acttcatgca gctgatccac gacgacagcc tgacctttaa agaggacatc cagaaagccc
     3541 aggtgtccgg ccagggcgat agcctgcacg agcacattgc caatctggcc ggcagccccg
     3601 ccattaagaa gggcatcctg cagacagtga aggtggtgga cgagctcgtg aaagtgatgg
     3661 gccggcacaa gcccgagaac atcgtgatcg aaatggccag agagaaccag accacccaga
     3721 agggacagaa gaacagccgc gagagaatga agcggatcga agagggcatc aaagagctgg
     3781 gcagccagat cctgaaagaa caccccgtgg aaaacaccca gctgcagaac gagaagctgt
     3841 acctgtacta cctgcagaat gggcgggata tgtacgtgga ccaggaactg gacatcaacc
     3901 ggctgtccga ctacgatgtg gaccatatcg tgcctcagag ctttctgaag gacgactcca
     3961 tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg caagagcgac aacgtgccct
     4021 ccgaagaggt cgtgaagaag atgaagaact actggcggca gctgctgaac gccaagctga
     4081 ttacccagag aaagttcgac aatctgacca aggccgagag aggcggcctg agcgaactgg
     4141 ataaggccgg cttcatcaag agacagctgg tggaaacccg gcagatcaca aagcacgtgg
     4201 cacagatcct ggactcccgg atgaacacta agtacgacga gaatgacaag ctgatccggg
     4261 aagtgaaagt gatcaccctg aagtccaagc tggtgtccga tttccggaag gatttccagt
     4321 tttacaaagt gcgcgagatc aacaactacc accacgccca cgacgcctac ctgaacgccg
     4381 tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg
     4441 actacaaggt gtacgacgtg cggaagatga tcgccaagag cgagcaggaa atcggcaagg
     4501 ctaccgccaa gtacttcttc tacagcaaca tcatgaactt tttcaagacc gagattaccc
     4561 tggccaacgg cgagatccgg aagcggcctc tgatcgagac aaacggcgaa accggggaga
     4621 tcgtgtggga taagggccgg gattttgcca ccgtgcggaa agtgctgagc atgccccaag
     4681 tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt cagcaaagag tctatcctgc
     4741 ccaagaggaa cagcgataag ctgatcgcca gaaagaagga ctgggaccct aagaagtacg
     4801 gcggcttcgc cagacccacc gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg
     4861 gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc atggaaagaa
     4921 gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa gaagtgaaaa
     4981 aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac ggccggaaga
     5041 gaatgctggc ctctgccggc atcctgcaga agggaaacga actggccctg ccctccaaat
     5101 atgtgaactt cctgtacctg gccagccact atgagaagct gaagggctcc cccgaggata
     5161 atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag atcatcgagc
     5221 agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac aaagtgctgt
     5281 ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat atcatccacc
     5341 tgtttaccct gaccaatctg ggagcccctg ccgccttcaa gtactttgac accaccatcg
     5401 accggaagat gtacagaagc accaaagagg tgctggacgc caccctgatc caccagagca
     5461 tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggtgac agcggcggga
     5521 gcggcgggag cggggggagc actaatctga gcgacatcat tgagaaggag actgggaaac
     5581 agctggtcat tcaggagtcc atcctgatgc tgcctgagga ggtggaggaa gtgatcggca
     5641 acaagccaga gtctgacatc ctggtgcaca ccgcctacga cgagtccaca gatgagaatg
     5701 tgatgctgct gacctctgac gcccccgagt ataagccttg ggccctggtc atccaggatt
     5761 ctaacggcga gaataagatc aagatgctga gcggaggatc cggaggatct ggaggcagca
     5821 ccaacctgtc tgacatcatc gagaaggaga caggcaagca gctggtcatc caggagagca
     5881 tcctgatgct gcccgaagaa gtcgaagaag tgatcggaaa caagcctgag agcgatatcc
     5941 tggtccatac cgcctacgac gagagtaccg acgaaaatgt gatgctgctg acatccgacg
     6001 ccccagagta taagccctgg gctctggtca tccaggattc caacggagag aacaaaatca
     6061 aaatgctgtc tggcggctca aaaagaaccg ccgacggcag cgaattcgag cccaagaaga
     6121 agaggaaagt cggaagcgga gctactaact tcagcctgct gaagcaggct ggagacgtgg
     6181 aggagaaccc tggacctatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     6241 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     6301 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     6361 ccgtgccctg gcccaccctc gtgaccaccc tgacctatgg agtgcagtgc ttcagccgct
     6421 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     6481 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     6541 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     6601 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     6661 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     6721 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     6781 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     6841 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     6901 acgagctgta caagtctggt ggttctccca agaagaagag gaaagtctaa ccggtcatca
     6961 tcaccatcac cattgagttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     7021 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     7081 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     7141 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     7201 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctcgat
     7261 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     7321 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagccta
     7381 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     7441 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     7501 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     7561 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     7621 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     7681 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     7741 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     7801 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     7861 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     7921 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     7981 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     8041 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     8101 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     8161 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     8221 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     8281 atctcaagaa gatcctttga tcttttctac ggggtctgac actcagtgga acgaaaactc
     8341 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     8401 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     8461 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     8521 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     8581 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     8641 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     8701 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     8761 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     8821 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     8881 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     8941 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     9001 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     9061 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     9121 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     9181 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     9241 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     9301 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     9361 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     9421 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc gatctcccga
     9481 tcccctaggg tcgactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     9541 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt a
//
