LOCUS       sequence-273619-        9553 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of AncBE4max-Cas9 Base-editor from an EF1a promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9553)
  AUTHORS   Standage-Beier K, Tekel SJ, Brookhouser N, Schwarz G, Nguyen T, Wang
            X, Brafman DA
  TITLE     A transient reporter for editing enrichment (TREE) in human cells.
  JOURNAL   Nucleic Acids Res. 2019 Aug 20. pii: 5552066. doi: 
            10.1093/nar/gkz713.
  PUBMED    31428784
REFERENCE   2  (bases 1 to 9553)
  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: "Nucleic 
            Acids Res. 2019 Aug 20. pii: 5552066. doi: 10.1093/nar/gkz713."
COMMENT     Sequence Label: pEF-AncBE4max
FEATURES             Location/Qualifiers
     source          1..9553
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        152..1334
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          383..1325
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1282..1302
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1344..1363
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1344..1362
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1412..1432
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2213..2236,2237..2239,2240..6313)
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             2213..6313
                     /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
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             6344..6592
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6623..6871
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6914..6934
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6943..6960
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6983..7000)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6989..7213
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7284..7300)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7284..7300)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7297..7319)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7308..7324
                     /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(7332..7338,7339..7356,7357..7362))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7377..7398
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7515..7532)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7686..8274)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7766..7785)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8445..9236,9237..9305))
                     /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     9068..9087
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9306..9410)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9489..9508)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
       61 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
      121 cgcgttgaca ttgattattg actagtcgtg aggctccggt gcccgtcagt gggcagagcg
      181 cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa ccggtgccta
      241 gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc
      301 cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa
      361 cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc ctggcctctt
      421 tacgggttat ggcccttgcg tgccttgaat tacttccacg cccctggctg cagtacgtga
      481 ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag
      541 gagccccttc gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc
      601 gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa
      661 atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc
      721 aagatctgca cactggtatt tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt
      781 cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga atcggacggg
      841 ggtagtctca agctggccgg cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc
      901 cgccctgggc ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc
      961 ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg
     1021 gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc agccgtcgct tcatgtgact
     1081 ccacggagta ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt tggagtacgt
     1141 cgtctttagg ttggggggag gggttttatg cgatggagtt tccccacact gagtgggtgg
     1201 agactgaagt taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg
     1261 agtttggatc ttggttcatt ctcaagcctc agacagtggt tcaaagtttt tttcttccat
     1321 ttcaggtgtc gtgaggcggc cgctaatacg actcactata gggagagccg ccaccatgaa
     1381 acggacagcc gacggaagcg agttcgagtc accaaagaag aagcggaaag tcagcagtga
     1441 aaccggacca gtggcagtgg acccaaccct gaggagacgg attgagcccc atgaatttga
     1501 agtgttcttt gacccaaggg agctgaggaa ggagacatgc ctgctgtacg agatcaagtg
     1561 gggcacaagc cacaagatct ggcgccacag ctccaagaac accacaaagc acgtggaagt
     1621 gaatttcatc gagaagttta cctccgagcg gcacttctgc ccctctacca gctgttccat
     1681 cacatggttt ctgtcttgga gcccttgcgg cgagtgttcc aaggccatca ccgagttcct
     1741 gtctcagcac cctaacgtga ccctggtcat ctacgtggcc cggctgtatc accacatgga
     1801 ccagcagaac aggcagggcc tgcgcgatct ggtgaattct ggcgtgacca tccagatcat
     1861 gacagcccca gagtacgact attgctggcg gaacttcgtg aattatccac ctggcaagga
     1921 ggcacactgg ccaagatacc cacccctgtg gatgaagctg tatgcactgg agctgcacgc
     1981 aggaatcctg ggcctgcctc catgtctgaa tatcctgcgg agaaagcagc cccagctgac
     2041 atttttcacc attgctctgc agtcttgtca ctatcagcgg ctgcctcctc atattctgtg
     2101 ggctacaggc ctgaagtctg gaggatctag cggaggatcc tctggcagcg agacaccagg
     2161 aacaagcgag tcagcaacac cagagagcag tggcggcagc agcggcggca gcgacaagaa
     2221 gtacagcatc ggcctggcca tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga
     2281 gtacaaggtg cccagcaaga aattcaaggt gctgggcaac accgaccggc acagcatcaa
     2341 gaagaacctg atcggagccc tgctgttcga cagcggcgaa acagccgagg ccacccggct
     2401 gaagagaacc gccagaagaa gatacaccag acggaagaac cggatctgct atctgcaaga
     2461 gatcttcagc aacgagatgg ccaaggtgga cgacagcttc ttccacagac tggaagagtc
     2521 cttcctggtg gaagaggata agaagcacga gcggcacccc atcttcggca acatcgtgga
     2581 cgaggtggcc taccacgaga agtaccccac catctaccac ctgagaaaga aactggtgga
     2641 cagcaccgac aaggccgacc tgcggctgat ctatctggcc ctggcccaca tgatcaagtt
     2701 ccggggccac ttcctgatcg agggcgacct gaaccccgac aacagcgacg tggacaagct
     2761 gttcatccag ctggtgcaga cctacaacca gctgttcgag gaaaacccca tcaacgccag
     2821 cggcgtggac gccaaggcca tcctgtctgc cagactgagc aagagcagac ggctggaaaa
     2881 tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg ttcggaaacc tgattgccct
     2941 gagcctgggc ctgaccccca acttcaagag caacttcgac ctggccgagg atgccaaact
     3001 gcagctgagc aaggacacct acgacgacga cctggacaac ctgctggccc agatcggcga
     3061 ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc gacgccatcc tgctgagcga
     3121 catcctgaga gtgaacaccg agatcaccaa ggcccccctg agcgcctcta tgatcaagag
     3181 atacgacgag caccaccagg acctgaccct gctgaaagct ctcgtgcggc agcagctgcc
     3241 tgagaagtac aaagagattt tcttcgacca gagcaagaac ggctacgccg gctacattga
     3301 cggcggagcc agccaggaag agttctacaa gttcatcaag cccatcctgg aaaagatgga
     3361 cggcaccgag gaactgctcg tgaagctgaa cagagaggac ctgctgcgga agcagcggac
     3421 cttcgacaac ggcagcatcc cccaccagat ccacctggga gagctgcacg ccattctgcg
     3481 gcggcaggaa gatttttacc cattcctgaa ggacaaccgg gaaaagatcg agaagatcct
     3541 gaccttccgc atcccctact acgtgggccc tctggccagg ggaaacagca gattcgcctg
     3601 gatgaccaga aagagcgagg aaaccatcac cccctggaac ttcgaggaag tggtggacaa
     3661 gggcgcttcc gcccagagct tcatcgagcg gatgaccaac ttcgataaga acctgcccaa
     3721 cgagaaggtg ctgcccaagc acagcctgct gtacgagtac ttcaccgtgt ataacgagct
     3781 gaccaaagtg aaatacgtga ccgagggaat gagaaagccc gccttcctga gcggcgagca
     3841 gaaaaaggcc atcgtggacc tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct
     3901 gaaagaggac tacttcaaga aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga
     3961 agatcggttc aacgcctccc tgggcacata ccacgatctg ctgaaaatta tcaaggacaa
     4021 ggacttcctg gacaatgagg aaaacgagga cattctggaa gatatcgtgc tgaccctgac
     4081 actgtttgag gacagagaga tgatcgagga acggctgaaa acctatgccc acctgttcga
     4141 cgacaaagtg atgaagcagc tgaagcggcg gagatacacc ggctggggca ggctgagccg
     4201 gaagctgatc aacggcatcc gggacaagca gtccggcaag acaatcctgg atttcctgaa
     4261 gtccgacggc ttcgccaaca gaaacttcat gcagctgatc cacgacgaca gcctgacctt
     4321 taaagaggac atccagaaag cccaggtgtc cggccagggc gatagcctgc acgagcacat
     4381 tgccaatctg gccggcagcc ccgccattaa gaagggcatc ctgcagacag tgaaggtggt
     4441 ggacgagctc gtgaaagtga tgggccggca caagcccgag aacatcgtga tcgaaatggc
     4501 cagagagaac cagaccaccc agaagggaca gaagaacagc cgcgagagaa tgaagcggat
     4561 cgaagagggc atcaaagagc tgggcagcca gatcctgaaa gaacaccccg tggaaaacac
     4621 ccagctgcag aacgagaagc tgtacctgta ctacctgcag aatgggcggg atatgtacgt
     4681 ggaccaggaa ctggacatca accggctgtc cgactacgat gtggaccata tcgtgcctca
     4741 gagctttctg aaggacgact ccatcgacaa caaggtgctg accagaagcg acaagaaccg
     4801 gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag aagatgaaga actactggcg
     4861 gcagctgctg aacgccaagc tgattaccca gagaaagttc gacaatctga ccaaggccga
     4921 gagaggcggc ctgagcgaac tggataaggc cggcttcatc aagagacagc tggtggaaac
     4981 ccggcagatc acaaagcacg tggcacagat cctggactcc cggatgaaca ctaagtacga
     5041 cgagaatgac aagctgatcc gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc
     5101 cgatttccgg aaggatttcc agttttacaa agtgcgcgag atcaacaact accaccacgc
     5161 ccacgacgcc tacctaaacg ccgtcgtggg aaccgccctg atcaaaaagt accctaagct
     5221 ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa
     5281 gagcgagcag gaaatcggca aggctaccgc caagtacttc ttctacagca acatcatgaa
     5341 ctttttcaag accgagatta ccctggccaa cggcgagatc cggaagcggc ctctgatcga
     5401 gacaaacggc gaaaccgggg agatcgtgtg ggataagggc cgggattttg ccaccgtgcg
     5461 gaaagtgctg agcatgcccc aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg
     5521 cttcagcaaa gagtctatcc tgcccaagag gaacagcgat aagctgatcg ccagaaagaa
     5581 ggactgggac cctaagaagt acggcggctt cgacagcccc accgtggcct attctgtgct
     5641 ggtggtggcc aaagtggaaa agggcaagtc caagaaactg aagagtgtga aagagctgct
     5701 ggggatcacc atcatggaaa gaagcagctt cgagaagaat cccatcgact ttctggaagc
     5761 caagggctac aaagaagtga aaaaggacct gatcatcaag ctgcctaagt actccctgtt
     5821 cgagctggaa aacggccgga agagaatgct ggcctctgcc ggcgaactgc agaagggaaa
     5881 cgaactggcc ctgccctcca aatatgtgaa cttcctgtac ctggccagcc actatgagaa
     5941 gctgaagggc tcccccgagg ataatgagca gaaacagctg tttgtggaac agcacaagca
     6001 ctacctggac gagatcatcg agcagatcag cgagttctcc aagagagtga tcctggccga
     6061 cgctaatctg gacaaagtgc tgtccgccta caacaagcac cgggataagc ccatcagaga
     6121 gcaggccgag aatatcatcc acctgtttac cctgaccaat ctgggagccc ctgccgcctt
     6181 caagtacttt gacaccacca tcgaccggaa gaggtacacc agcaccaaag aggtgctgga
     6241 cgccaccctg atccaccaga gcatcaccgg cctgtacgag acacggatcg acctgtctca
     6301 gctgggaggt gacagcggcg ggagcggcgg gagcgggggg agcactaatc tgagcgacat
     6361 cattgagaag gagactggga aacagctggt cattcaggag tccatcctga tgctgcctga
     6421 ggaggtggag gaagtgatcg gcaacaagcc agagtctgac atcctggtgc acaccgccta
     6481 cgacgagtcc acagatgaga atgtgatgct gctgacctct gacgcccccg agtataagcc
     6541 ttgggccctg gtcatccagg attctaacgg cgagaataag atcaagatgc tgagcggagg
     6601 atccggagga tctggaggca gcaccaacct gtctgacatc atcgagaagg agacaggcaa
     6661 gcagctggtc atccaggaga gcatcctgat gctgcccgaa gaagtcgaag aagtgatcgg
     6721 aaacaagcct gagagcgata tcctggtcca taccgcctac gacgagagta ccgacgaaaa
     6781 tgtgatgctg ctgacatccg acgccccaga gtataagccc tgggctctgg tcatccagga
     6841 ttccaacgga gagaacaaaa tcaaaatgct gtctggcggc tcaaaaagaa ccgccgacgg
     6901 cagcgaattc gagcccaaga agaagaggaa agtctaaccg gtcatcatca ccatcaccat
     6961 tgagtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     7021 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     7081 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     7141 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     7201 gtgggctcta tggcttctga ggcggaaaga accagctggg gctcgatacc gtcgacctct
     7261 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     7321 acaattccac acaacatacg agccggaagc ataaagtgta aagcctagga tgcctaatga
     7381 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     7441 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggaa gaggcggttt gcgtattggg
     7501 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     7561 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     7621 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     7681 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     7741 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     7801 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     7861 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     7921 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     7981 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     8041 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     8101 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     8161 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     8221 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     8281 cctttgatct tttctacggg gtctgacact cagtggaacg aaaactcacg ttaagggatt
     8341 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     8401 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     8461 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     8521 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     8581 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     8641 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     8701 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     8761 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     8821 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     8881 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     8941 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     9001 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     9061 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     9121 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     9181 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     9241 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     9301 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     9361 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     9421 cgaaaagtgc cacctgacgt cgacggatcg ggagatcgat ctcccgatcc cctagggtcg
     9481 actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct ccctgcttgt
     9541 gtgttggagg tcg
//
