LOCUS       sequence-269626-        8961 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses BE4max-NRCH in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8961)
  AUTHORS   Miller SM, Wang T, Randolph PB, Arbab M, Shen MW, Huang TP, Matuszek
            Z, Newby GA, Rees HA, Liu DR
  TITLE     Continuous evolution of SpCas9 variants compatible with non-G PAMs.
  JOURNAL   Nat Biotechnol. 2020 Apr;38(4):471-481. doi: 
            10.1038/s41587-020-0412-8. Epub 2020 Feb 10.
  PUBMED    32042170
REFERENCE   2  (bases 1 to 8961)
  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"; doi: 
            "10.1038/s41587-020-0412-8"; journalName: "Nat Biotechnol"; date: 
            "2020-04"; volume: "38"; issue: "4"; pages: "471-481"
COMMENT     Sequence Label: pCMV-BE4max-NRCH
FEATURES             Location/Qualifiers
     source          1..8961
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     246..265
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        246..264
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             314..334
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             335..1018
                     /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             1115..5215
                     /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="DKKYSIGLTIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMVKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGIIPHQIHLGELHAILRRQGDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRLRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGGH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKGNSDKLIARKKDWDPKKYGGFNSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGVLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTINRKQYNTTKEVLD
                     ATLIRQSITGLYETRIDLSQLGGD"
     CDS             5246..5494
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5525..5773
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5816..5836
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5845..5862
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5885..5902)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5891..6115
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6186..6202)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6186..6202)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6199..6221)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6210..6226
                     /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(6234..6240,6241..6258,6259..6264))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6279..6300
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6417..6434)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6588..7176)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6668..6687)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7347..8138,8139..8207))
                     /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     7970..7989
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8208..8312)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8391..8410)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        8582..8961
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gagatccgcg
      241 gccgctaata cgactcacta tagggagagc cgccaccatg aaacggacag ccgacggaag
      301 cgagttcgag tcaccaaaga agaagcggaa agtctcctca gagactgggc ctgtcgccgt
      361 cgatccaacc ctgcgccgcc ggattgaacc tcacgagttt gaagtgttct ttgacccccg
      421 ggagctgaga aaggagacat gcctgctgta cgagatcaac tggggaggca ggcactccat
      481 ctggaggcac acctctcaga acacaaataa gcacgtggag gtgaacttca tcgagaagtt
      541 taccacagag cggtacttct gccccaatac cagatgtagc atcacatggt ttctgagctg
      601 gtccccttgc ggagagtgta gcagggccat caccgagttc ctgtccagat atccacacgt
      661 gacactgttt atctacatcg ccaggctgta tcaccacgca gacccaagga ataggcaggg
      721 cctgcgcgat ctgatcagct ccggcgtgac catccagatc atgacagagc aggagtccgg
      781 ctactgctgg cggaacttcg tgaattattc tcctagcaac gaggcccact ggcctaggta
      841 cccacacctg tgggtgcgcc tgtacgtgct ggagctgtat tgcatcatcc tgggcctgcc
      901 cccttgtctg aatatcctgc ggagaaagca gccccagctg accttcttta caatcgccct
      961 gcagtcttgt cactatcaga ggctgccacc ccacatcctg tgggccacag gcctgaagtc
     1021 tggaggatct agcggaggat cctctggcag cgagacacca ggaacaagcg agtcagcaac
     1081 accagagagc agtggcggca gcagcggcgg cagcgacaag aagtacagca tcggcctgac
     1141 catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg tgcccagcaa
     1201 gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc tgatcggagc
     1261 cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa ccgccagaag
     1321 aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca gcaacgagat
     1381 ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg tggaagagga
     1441 taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg cctaccacga
     1501 gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg acaaggccga
     1561 cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc acttcctgat
     1621 cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc agctggtgca
     1681 gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc
     1741 catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg cccagctgcc
     1801 cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc
     1861 caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac
     1921 ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt
     1981 tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac
     2041 cgagatcacc aaggcccccc tgagcgcctc tatggtgaag agatacgacg agcaccacca
     2101 ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat
     2161 tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga
     2221 agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct
     2281 cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcattat
     2341 cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg gcgattttta
     2401 cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta
     2461 ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga
     2521 ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag
     2581 cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa
     2641 gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt
     2701 gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga
     2761 cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa
     2821 gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc
     2881 cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga
     2941 ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga
     3001 gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca
     3061 gctgaagcgg ctgagataca ccggctgggg caggctgagc cggaagctga tcaacggcat
     3121 ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa
     3181 cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa
     3241 agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag
     3301 ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt
     3361 gatgggcggc cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac
     3421 ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga
     3481 gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa
     3541 gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat
     3601 caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc tgaaggacga
     3661 ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt
     3721 gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa
     3781 gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga
     3841 actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca
     3901 cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat
     3961 ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt
     4021 ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa
     4081 cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta
     4141 cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg
     4201 caaggctacc gccaagtact tcttctacag caacatcatg aactttttca agaccgagat
     4261 taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg
     4321 ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc
     4381 ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat
     4441 cctgcccaag ggtaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa
     4501 gtacggcggc ttcaacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga
     4561 aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga
     4621 aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt
     4681 gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg
     4741 gaagagaatg ctggcctctg ccggcgtgct gcagaaggga aacgaactgg ccctgccctc
     4801 caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga
     4861 ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat
     4921 cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt
     4981 gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat
     5041 ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac
     5101 catcaaccgg aagcaataca acacgaccaa agaggtgctg gacgccaccc tgatccgtca
     5161 gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag gtgacagcgg
     5221 cgggagcggc gggagcgggg ggagcactaa tctgagcgac atcattgaga aggagactgg
     5281 gaaacagctg gtcattcagg agtccatcct gatgctgcct gaggaggtgg aggaagtgat
     5341 cggcaacaag ccagagtctg acatcctggt gcacaccgcc tacgacgagt ccacagatga
     5401 gaatgtgatg ctgctgacct ctgacgcccc cgagtataag ccttgggccc tggtcatcca
     5461 ggattctaac ggcgagaata agatcaagat gctgagcgga ggatccggag gatctggagg
     5521 cagcaccaac ctgtctgaca tcatcgagaa ggagacaggc aagcagctgg tcatccagga
     5581 gagcatcctg atgctgcccg aagaagtcga agaagtgatc ggaaacaagc ctgagagcga
     5641 tatcctggtc cataccgcct acgacgagag taccgacgaa aatgtgatgc tgctgacatc
     5701 cgacgcccca gagtataagc cctgggctct ggtcatccag gattccaacg gagagaacaa
     5761 aatcaaaatg ctgtctggcg gctcaaaaag aaccgccgac ggcagcgaat tcgagcccaa
     5821 gaagaagagg aaagtctaac cggtcatcat caccatcacc attgagttta aacccgctga
     5881 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     5941 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     6001 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     6061 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     6121 gaggcggaaa gaaccagctg gggctcaata ccgtcgacct ctagctagag cttggcgtaa
     6181 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     6241 cgagccggaa gcataaagtg taaagcctag ggtgcctaat gagtgagcta actcacatta
     6301 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     6361 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6421 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6481 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6541 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6601 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6661 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6721 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6781 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     6841 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     6901 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     6961 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     7021 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     7081 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     7141 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     7201 gggtctgaca ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     7261 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     7321 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7381 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7441 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7501 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7561 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7621 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7681 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7741 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7801 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7861 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7921 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7981 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     8041 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     8101 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     8161 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     8221 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     8281 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     8341 gtcgacggat cgggagatcg atctcccgat cccctagggt cgactctcag tacaatctgc
     8401 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     8461 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     8521 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt
     8581 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
     8641 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     8701 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     8761 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     8821 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     8881 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     8941 ttagtcatcg ctattaccat g
//
