LOCUS       sequence-266849-        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        12..1194
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          243..1185
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1142..1162
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1204..1223
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1204..1222
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1272..1292
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(2073..2096,2097..2099,2100..6173)
                     /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             2073..6173
                     /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             6204..6452
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6483..6731
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6774..6794
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6803..6820
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6843..6860)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6849..7073
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7144..7160)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7144..7160)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7157..7179)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7168..7184
                     /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(7192..7198,7199..7216,7217..7222))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7237..7258
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7375..7392)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7546..8134)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7626..7645)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8305..9096,9097..9165))
                     /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     8928..8947
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9166..9270)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9349..9368)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 actagtcgtg aggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg
       61 agaagttggg gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa
      121 actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt
      181 atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac
      241 aggtaagtgc cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg
      301 tgccttgaat tacttccacg cccctggctg cagtacgtga ttcttgatcc cgagcttcgg
      361 gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct
      421 tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc
      481 gcctgtctcg ctgctttcga taagtctcta gccatttaaa atttttgatg acctgctgcg
      541 acgctttttt tctggcaaga tagtcttgta aatgcgggcc aagatctgca cactggtatt
      601 tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg
      661 aggcggggcc tgcgagcgcg gccaccgaga atcggacggg ggtagtctca agctggccgg
      721 cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg
      781 gcccggtcgg caccagttgc gtgagcggaa agatggccgc ttcccggccc tgctgcaggg
      841 agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg
      901 aaaagggcct ttccgtcctc agccgtcgct tcatgtgact ccacggagta ccgggcgccg
      961 tccaggcacc tcgattagtt ctcgagcttt tggagtacgt cgtctttagg ttggggggag
     1021 gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt taggccagct
     1081 tggcacttga tgtaattctc cttggaattt gccctttttg agtttggatc ttggttcatt
     1141 ctcaagcctc agacagtggt tcaaagtttt tttcttccat ttcaggtgtc gtgaggcggc
     1201 cgctaatacg actcactata gggagagccg ccaccatgaa acggacagcc gacggaagcg
     1261 agttcgagtc accaaagaag aagcggaaag tcagcagtga aaccggacca gtggcagtgg
     1321 acccaaccct gaggagacgg attgagcccc atgaatttga agtgttcttt gacccaaggg
     1381 agctgaggaa ggagacatgc ctgctgtacg agatcaagtg gggcacaagc cacaagatct
     1441 ggcgccacag ctccaagaac accacaaagc acgtggaagt gaatttcatc gagaagttta
     1501 cctccgagcg gcacttctgc ccctctacca gctgttccat cacatggttt ctgtcttgga
     1561 gcccttgcgg cgagtgttcc aaggccatca ccgagttcct gtctcagcac cctaacgtga
     1621 ccctggtcat ctacgtggcc cggctgtatc accacatgga ccagcagaac aggcagggcc
     1681 tgcgcgatct ggtgaattct ggcgtgacca tccagatcat gacagcccca gagtacgact
     1741 attgctggcg gaacttcgtg aattatccac ctggcaagga ggcacactgg ccaagatacc
     1801 cacccctgtg gatgaagctg tatgcactgg agctgcacgc aggaatcctg ggcctgcctc
     1861 catgtctgaa tatcctgcgg agaaagcagc cccagctgac atttttcacc attgctctgc
     1921 agtcttgtca ctatcagcgg ctgcctcctc atattctgtg ggctacaggc ctgaagtctg
     1981 gaggatctag cggaggatcc tctggcagcg agacaccagg aacaagcgag tcagcaacac
     2041 cagagagcag tggcggcagc agcggcggca gcgacaagaa gtacagcatc ggcctggcca
     2101 tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg cccagcaaga
     2161 aattcaaggt gctgggcaac accgaccggc acagcatcaa gaagaacctg atcggagccc
     2221 tgctgttcga cagcggcgaa acagccgagg ccacccggct gaagagaacc gccagaagaa
     2281 gatacaccag acggaagaac cggatctgct atctgcaaga gatcttcagc aacgagatgg
     2341 ccaaggtgga cgacagcttc ttccacagac tggaagagtc cttcctggtg gaagaggata
     2401 agaagcacga gcggcacccc atcttcggca acatcgtgga cgaggtggcc taccacgaga
     2461 agtaccccac catctaccac ctgagaaaga aactggtgga cagcaccgac aaggccgacc
     2521 tgcggctgat ctatctggcc ctggcccaca tgatcaagtt ccggggccac ttcctgatcg
     2581 agggcgacct gaaccccgac aacagcgacg tggacaagct gttcatccag ctggtgcaga
     2641 cctacaacca gctgttcgag gaaaacccca tcaacgccag cggcgtggac gccaaggcca
     2701 tcctgtctgc cagactgagc aagagcagac ggctggaaaa tctgatcgcc cagctgcccg
     2761 gcgagaagaa gaatggcctg ttcggaaacc tgattgccct gagcctgggc ctgaccccca
     2821 acttcaagag caacttcgac ctggccgagg atgccaaact gcagctgagc aaggacacct
     2881 acgacgacga cctggacaac ctgctggccc agatcggcga ccagtacgcc gacctgtttc
     2941 tggccgccaa gaacctgtcc gacgccatcc tgctgagcga catcctgaga gtgaacaccg
     3001 agatcaccaa ggcccccctg agcgcctcta tgatcaagag atacgacgag caccaccagg
     3061 acctgaccct gctgaaagct ctcgtgcggc agcagctgcc tgagaagtac aaagagattt
     3121 tcttcgacca gagcaagaac ggctacgccg gctacattga cggcggagcc agccaggaag
     3181 agttctacaa gttcatcaag cccatcctgg aaaagatgga cggcaccgag gaactgctcg
     3241 tgaagctgaa cagagaggac ctgctgcgga agcagcggac cttcgacaac ggcagcatcc
     3301 cccaccagat ccacctggga gagctgcacg ccattctgcg gcggcaggaa gatttttacc
     3361 cattcctgaa ggacaaccgg gaaaagatcg agaagatcct gaccttccgc atcccctact
     3421 acgtgggccc tctggccagg ggaaacagca gattcgcctg gatgaccaga aagagcgagg
     3481 aaaccatcac cccctggaac ttcgaggaag tggtggacaa gggcgcttcc gcccagagct
     3541 tcatcgagcg gatgaccaac ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc
     3601 acagcctgct gtacgagtac ttcaccgtgt ataacgagct gaccaaagtg aaatacgtga
     3661 ccgagggaat gagaaagccc gccttcctga gcggcgagca gaaaaaggcc atcgtggacc
     3721 tgctgttcaa gaccaaccgg aaagtgaccg tgaagcagct gaaagaggac tacttcaaga
     3781 aaatcgagtg cttcgactcc gtggaaatct ccggcgtgga agatcggttc aacgcctccc
     3841 tgggcacata ccacgatctg ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg
     3901 aaaacgagga cattctggaa gatatcgtgc tgaccctgac actgtttgag gacagagaga
     3961 tgatcgagga acggctgaaa acctatgccc acctgttcga cgacaaagtg atgaagcagc
     4021 tgaagcggcg gagatacacc ggctggggca ggctgagccg gaagctgatc aacggcatcc
     4081 gggacaagca gtccggcaag acaatcctgg atttcctgaa gtccgacggc ttcgccaaca
     4141 gaaacttcat gcagctgatc cacgacgaca gcctgacctt taaagaggac atccagaaag
     4201 cccaggtgtc cggccagggc gatagcctgc acgagcacat tgccaatctg gccggcagcc
     4261 ccgccattaa gaagggcatc ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga
     4321 tgggccggca caagcccgag aacatcgtga tcgaaatggc cagagagaac cagaccaccc
     4381 agaagggaca gaagaacagc cgcgagagaa tgaagcggat cgaagagggc atcaaagagc
     4441 tgggcagcca gatcctgaaa gaacaccccg tggaaaacac ccagctgcag aacgagaagc
     4501 tgtacctgta ctacctgcag aatgggcggg atatgtacgt ggaccaggaa ctggacatca
     4561 accggctgtc cgactacgat gtggaccata tcgtgcctca gagctttctg aaggacgact
     4621 ccatcgacaa caaggtgctg accagaagcg acaagaaccg gggcaagagc gacaacgtgc
     4681 cctccgaaga ggtcgtgaag aagatgaaga actactggcg gcagctgctg aacgccaagc
     4741 tgattaccca gagaaagttc gacaatctga ccaaggccga gagaggcggc ctgagcgaac
     4801 tggataaggc cggcttcatc aagagacagc tggtggaaac ccggcagatc acaaagcacg
     4861 tggcacagat cctggactcc cggatgaaca ctaagtacga cgagaatgac aagctgatcc
     4921 gggaagtgaa agtgatcacc ctgaagtcca agctggtgtc cgatttccgg aaggatttcc
     4981 agttttacaa agtgcgcgag atcaacaact accaccacgc ccacgacgcc tacctaaacg
     5041 ccgtcgtggg aaccgccctg atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg
     5101 gcgactacaa ggtgtacgac gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca
     5161 aggctaccgc caagtacttc ttctacagca acatcatgaa ctttttcaag accgagatta
     5221 ccctggccaa cggcgagatc cggaagcggc ctctgatcga gacaaacggc gaaaccgggg
     5281 agatcgtgtg ggataagggc cgggattttg ccaccgtgcg gaaagtgctg agcatgcccc
     5341 aagtgaatat cgtgaaaaag accgaggtgc agacaggcgg cttcagcaaa gagtctatcc
     5401 tgcccaagag gaacagcgat aagctgatcg ccagaaagaa ggactgggac cctaagaagt
     5461 acggcggctt cgacagcccc accgtggcct attctgtgct ggtggtggcc aaagtggaaa
     5521 agggcaagtc caagaaactg aagagtgtga aagagctgct ggggatcacc atcatggaaa
     5581 gaagcagctt cgagaagaat cccatcgact ttctggaagc caagggctac aaagaagtga
     5641 aaaaggacct gatcatcaag ctgcctaagt actccctgtt cgagctggaa aacggccgga
     5701 agagaatgct ggcctctgcc ggcgaactgc agaagggaaa cgaactggcc ctgccctcca
     5761 aatatgtgaa cttcctgtac ctggccagcc actatgagaa gctgaagggc tcccccgagg
     5821 ataatgagca gaaacagctg tttgtggaac agcacaagca ctacctggac gagatcatcg
     5881 agcagatcag cgagttctcc aagagagtga tcctggccga cgctaatctg gacaaagtgc
     5941 tgtccgccta caacaagcac cgggataagc ccatcagaga gcaggccgag aatatcatcc
     6001 acctgtttac cctgaccaat ctgggagccc ctgccgcctt caagtacttt gacaccacca
     6061 tcgaccggaa gaggtacacc agcaccaaag aggtgctgga cgccaccctg atccaccaga
     6121 gcatcaccgg cctgtacgag acacggatcg acctgtctca gctgggaggt gacagcggcg
     6181 ggagcggcgg gagcgggggg agcactaatc tgagcgacat cattgagaag gagactggga
     6241 aacagctggt cattcaggag tccatcctga tgctgcctga ggaggtggag gaagtgatcg
     6301 gcaacaagcc agagtctgac atcctggtgc acaccgccta cgacgagtcc acagatgaga
     6361 atgtgatgct gctgacctct gacgcccccg agtataagcc ttgggccctg gtcatccagg
     6421 attctaacgg cgagaataag atcaagatgc tgagcggagg atccggagga tctggaggca
     6481 gcaccaacct gtctgacatc atcgagaagg agacaggcaa gcagctggtc atccaggaga
     6541 gcatcctgat gctgcccgaa gaagtcgaag aagtgatcgg aaacaagcct gagagcgata
     6601 tcctggtcca taccgcctac gacgagagta ccgacgaaaa tgtgatgctg ctgacatccg
     6661 acgccccaga gtataagccc tgggctctgg tcatccagga ttccaacgga gagaacaaaa
     6721 tcaaaatgct gtctggcggc tcaaaaagaa ccgccgacgg cagcgaattc gagcccaaga
     6781 agaagaggaa agtctaaccg gtcatcatca ccatcaccat tgagtttaaa cccgctgatc
     6841 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     6901 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     6961 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     7021 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     7081 ggcggaaaga accagctggg gctcgatacc gtcgacctct agctagagct tggcgtaatc
     7141 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     7201 agccggaagc ataaagtgta aagcctagga tgcctaatga gtgagctaac tcacattaat
     7261 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     7321 aatcggccaa cgcgcgggaa gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     7381 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     7441 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     7501 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     7561 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     7621 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     7681 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     7741 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     7801 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     7861 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     7921 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     7981 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     8041 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     8101 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     8161 gtctgacact cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     8221 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     8281 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     8341 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     8401 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     8461 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     8521 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     8581 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     8641 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     8701 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     8761 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     8821 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     8881 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     8941 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     9001 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     9061 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     9121 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     9181 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     9241 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     9301 cgacggatcg ggagatcgat ctcccgatcc cctagggtcg actctcagta caatctgctc
     9361 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
     9421 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
     9481 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg
     9541 acattgatta ttg
//
