LOCUS       sequence-309498-       10437 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T7 promoter bacterial expression plasmid for hAPOBEC3A(N57Q)-BE3 
            with N-terminal 6xHis-tag and C-terminal SV40 NLS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10437)
  AUTHORS   Zeng J, Wu Y, Ren C, Bonanno J, Shen AH, Shea D, Gehrke JM, Clement 
            K, Luk K, Yao Q, Kim R, Wolfe SA, Manis JP, Pinello L, Joung JK, 
            Bauer DE
  TITLE     Therapeutic base editing of human hematopoietic stem cells.
  JOURNAL   Nat Med. 2020 Apr;26(4):535-541. doi: 10.1038/s41591-020-0790-y. 
            Epub 2020 Mar 16.
  PUBMED    32284612
REFERENCE   2  (bases 1 to 10437)
  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/s41591-020-0790-y"; journalName: "Nat Med"; date: 
            "2020-04"; volume: "26"; issue: "4"; pages: "535-541"
COMMENT     Sequence Label: pET-A3A(N57Q)-BE3
FEATURES             Location/Qualifiers
     source          1..10437
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     220..239
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     473..490
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    505..644
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(570..589)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     730..752
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(746..937)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1746..2828)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     2790..2809
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2829..2906)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3112..3131
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3215..3234
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3215..3233
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3234..3258
                     /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)."
     RBS             3273..3295
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3314..3331
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(3570..3581)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             join(3977..4000,4001..4003,4004..8077)
                     /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             8090..8338
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             8351..8371
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8380..8397
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             8516..8533
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      8600..8647
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      8684..9139
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8771..8790)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8981..9002
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(9231..10046)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(9231..10046)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     9954..9973
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     10038..10057
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(10168..10437,1..319)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
       61 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
      121 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
      181 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
      241 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
      301 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
      361 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
      421 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
      481 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
      541 tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
      601 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
      661 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
      721 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
      781 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt
      841 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc
      901 gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat
      961 ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta
     1021 ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga
     1081 tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg
     1141 taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc
     1201 agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg
     1261 ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg
     1321 gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca
     1381 ggagcacgat catgcgcacc cgtggggccg ccatgccggc gataatggcc tgcttctcgc
     1441 cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga
     1501 ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa
     1561 tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa
     1621 gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc
     1681 tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt
     1741 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     1801 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc
     1861 agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag
     1921 cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg
     1981 atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg
     2041 cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc
     2101 agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac
     2161 atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat
     2221 ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc
     2281 gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca
     2341 tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga
     2401 acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg
     2461 atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg
     2521 ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta
     2581 atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc
     2641 agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc
     2701 gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc
     2761 acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact
     2821 ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga
     2881 aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg
     2941 cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg
     3001 tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac
     3061 gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc
     3121 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc
     3181 ggcgtagagg atcgagatct cgatcccgcg aaattaatac gactcactat aggggaattg
     3241 tgagcggata acaattcccc tctagaaata attttgttta actttaagaa ggagatatac
     3301 catgggcagc agccatcatc atcatcatca catggaggct agtccagcct ctggcccacg
     3361 tcatttaatg gatcctcata tctttactag caactttaat aacggcatcg gccgtcacaa
     3421 gacgtacctt tgctatgagg ttgagcgttt agacaatggc acatcagtta agatggacca
     3481 gcatcgcggc ttcttgcacc aacaagcaaa gaaccttttg tgtggctttt atggtcgtca
     3541 tgcagagtta cgctttctgg atcttgtccc ctcccttcaa ttagatccgg ctcagatcta
     3601 tcgcgtcacc tggtttattt catggagtcc gtgttttagt tggggttgtg caggagaggt
     3661 tcgtgcattt cttcaagaaa atacgcatgt gcgtttacgt atttttgcgg ctcgtattta
     3721 tgactacgat ccactttaca aagaagctct tcagatgtta cgtgacgcag gtgcccaggt
     3781 ctcgattatg acttatgacg agtttaagca ttgttgggac acttttgtag atcaccaagg
     3841 ttgcccattt caaccatggg acgggttgga tgagcacagt caggctctta gcggacgctt
     3901 acgtgcgatc ttgcagaacc aggggaactc cggtagcgaa acaccgggga cttcagaatc
     3961 ggccaccccg gagtctgata agaaatactc aataggctta gctatcggca caaatagcgt
     4021 cggatgggcg gtgatcactg atgaatataa ggttccgtct aaaaagttca aggttctggg
     4081 aaatacagac cgccacagta tcaaaaaaaa tcttataggg gctcttttat ttgacagtgg
     4141 agagacagcg gaagcgactc gtctcaaacg gacagctcgt agaaggtata cacgtcggaa
     4201 gaatcgtatt tgttatctac aggagatttt ttcaaatgag atggcgaaag tagatgatag
     4261 tttctttcat cgacttgaag agtctttttt ggtggaagaa gacaagaagc atgaacgtca
     4321 tcctattttt ggaaatatag tagatgaagt tgcttatcat gagaaatatc caactatcta
     4381 tcatctgcga aaaaaattgg tagattctac tgataaagcg gatttgcgct taatctattt
     4441 ggccttagcg catatgatta agtttcgtgg tcattttttg attgagggag atttaaatcc
     4501 tgataatagt gatgtggaca aactatttat ccagttggta caaacctaca atcaattatt
     4561 tgaagaaaac cctattaacg caagtggagt agatgctaaa gcgattcttt ctgcacgatt
     4621 gagtaaatca agacgattag aaaatctcat tgctcagctc cccggtgaga agaaaaatgg
     4681 cttatttggg aatctcattg ctttgtcatt gggtttgacc cctaatttta aatcaaattt
     4741 tgatttggca gaagatgcta aattacagct ttcaaaagat acttacgatg atgatttaga
     4801 taatttattg gcgcaaattg gagatcaata tgctgatttg tttttggcag ctaagaattt
     4861 atcagatgct attttacttt cagatatcct aagagtaaat actgaaataa ctaaggctcc
     4921 cctatcagct tcaatgatta aacgctacga tgaacatcat caagacttga ctcttttaaa
     4981 agctttagtt cgacaacaac ttccagaaaa gtataaagaa atcttttttg atcaatcaaa
     5041 aaacggatat gcaggttata ttgatggggg agctagccaa gaagaatttt ataaatttat
     5101 caaaccaatt ttagaaaaaa tggatggtac tgaggaatta ttggtgaaac taaatcgtga
     5161 agatttgctg cgcaagcaac ggacctttga caacggctct attccccatc aaattcactt
     5221 gggtgagctg catgctattt tgagaagaca agaagacttt tatccatttt taaaagacaa
     5281 tcgtgagaag attgaaaaaa tcttgacttt tcgaattcct tattatgttg gtccattggc
     5341 gcgtggcaat agtcgttttg catggatgac tcggaagtct gaagaaacaa ttaccccatg
     5401 gaattttgaa gaagttgtcg ataaaggtgc ttcagctcaa tcatttattg aacgcatgac
     5461 aaactttgat aaaaatcttc caaatgaaaa agtactacca aaacatagtt tgctttatga
     5521 gtattttacg gtttataacg aattgacaaa ggtcaaatat gttactgaag gaatgcgaaa
     5581 accagcattt ctttcaggtg aacagaagaa agccattgtt gatttactct tcaaaacaaa
     5641 tcgaaaagta accgttaagc aattaaaaga agattatttc aaaaaaatag aatgttttga
     5701 tagtgttgaa atttcaggag ttgaagatag atttaatgct tcattaggta cctaccatga
     5761 tttgctaaaa attattaaag ataaagattt tttggataat gaagaaaatg aagatatctt
     5821 agaggatatt gttttaacat tgaccttatt tgaagatagg gagatgattg aggaaagact
     5881 taaaacatat gctcacctct ttgatgataa ggtgatgaaa cagcttaaac gtcgccgtta
     5941 tactggttgg ggacgtttgt ctcgaaaatt gattaatggt attagggata agcaatctgg
     6001 caaaacaata ttagattttt tgaaatcaga tggttttgcc aatcgcaatt ttatgcagct
     6061 gatccatgat gatagtttga catttaaaga agacattcaa aaagcacaag tgtctggaca
     6121 aggcgatagt ttacatgaac atattgcaaa tttagctggt agccctgcta ttaaaaaagg
     6181 tattttacag actgtaaaag ttgttgatga attggtcaaa gtaatggggc ggcataagcc
     6241 agaaaatatc gttattgaaa tggcacgtga aaatcagaca actcaaaagg gccagaaaaa
     6301 ttcgcgagag cgtatgaaac gaatcgaaga aggtatcaaa gaattaggaa gtcagattct
     6361 taaagagcat cctgttgaaa atactcaatt gcaaaatgaa aagctctatc tctattatct
     6421 ccaaaatgga agagacatgt atgtggacca agaattagat attaatcgtt taagtgatta
     6481 tgatgtcgat cacattgttc cacaaagttt ccttaaagac gattcaatag acaataaggt
     6541 cttaacgcgt tctgataaaa atcgtggtaa atcggataac gttccaagtg aagaagtagt
     6601 caaaaagatg aaaaactatt ggagacaact tctaaacgcc aagttaatca ctcaacgtaa
     6661 gtttgataat ttaacgaaag ctgaacgtgg aggtttgagt gaacttgata aagctggttt
     6721 tatcaaacgc caattggttg aaactcgcca aatcactaag catgtggcac aaattttgga
     6781 tagtcgcatg aatactaaat acgatgaaaa tgataaactt attcgagagg ttaaagtgat
     6841 taccttaaaa tctaaattag tttctgactt ccgaaaagat ttccaattct ataaagtacg
     6901 tgagattaac aattaccatc atgcccatga tgcgtatcta aatgccgtcg ttggaactgc
     6961 tttgattaag aaatatccaa aacttgaatc ggagtttgtc tatggtgatt ataaagttta
     7021 tgatgttcgt aaaatgattg ctaagtctga gcaagaaata ggcaaagcaa ccgcaaaata
     7081 tttcttttac tctaatatca tgaacttctt caaaacagaa attacacttg caaatggaga
     7141 gattcgcaaa cgccctctaa tcgaaactaa tggggaaact ggagaaattg tctgggataa
     7201 agggcgagat tttgccacag tgcgcaaagt attgtccatg ccccaagtca atattgtcaa
     7261 gaaaacagaa gtacagacag gcggattctc caaggagtca attttaccaa aaagaaattc
     7321 ggacaagctt attgctcgta aaaaagactg ggatccaaaa aaatatggtg gttttgatag
     7381 tccaacggta gcttattcag tcctagtggt tgctaaggtg gaaaaaggga aatcgaagaa
     7441 gttaaaatcc gttaaagagt tactagggat cacaattatg gaaagaagtt cctttgaaaa
     7501 aaatccgatt gactttttag aagctaaagg atataaggaa gttaaaaaag acttaatcat
     7561 taaactacct aaatatagtc tttttgagtt agaaaacggt cgtaaacgga tgctggctag
     7621 tgccggagaa ttacaaaaag gaaatgagct ggctctgcca agcaaatatg tgaatttttt
     7681 atatttagct agtcattatg aaaagttgaa gggtagtcca gaagataacg aacaaaaaca
     7741 attgtttgtg gagcagcata agcattattt agatgagatt attgagcaaa tcagtgaatt
     7801 ttctaagcgt gttattttag cagatgccaa tttagataaa gttcttagtg catataacaa
     7861 acatagagac aaaccaatac gtgaacaagc agaaaatatt attcatttat ttacgttgac
     7921 gaatcttgga gctccagccg cattcaagta ttttgacaca acgatagatc gcaaacgata
     7981 cacttctacc aaggaggtgc tagacgcgac actgattcac caatccatca cgggattata
     8041 tgaaactcgg atagatttgt cacagcttgg gggtgactct ggtggttcta ctaatctgtc
     8101 agatattatt gaaaaggaga ccggtaagca actggttatc caggaatcca tcctcatgct
     8161 cccagaggag gtggaagaag tcattgggaa caagccggaa agcgatatac tcgtgcacac
     8221 cgcctacgac gagagcaccg acgagaatgt catgcttctg actagcgacg cccctgaata
     8281 caagccttgg gctctggtca tacaggatag caacggtgag aacaagatta agatgctctc
     8341 tggtggttct cccaagaaga agaggaaagt ctaatcgagc accaccacca ccaccactga
     8401 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
     8461 taactagcat aaccccttgg ggagctccgt cgacaagctt gcggccgcac tcgagcacca
     8521 ccaccaccac cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     8581 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     8641 ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc
     8701 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     8761 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     8821 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     8881 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     8941 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     9001 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     9061 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     9121 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     9181 gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc ttagaaaaac
     9241 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
     9301 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
     9361 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
     9421 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
     9481 gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc
     9541 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
     9601 agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
     9661 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
     9721 acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta
     9781 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
     9841 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
     9901 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga
     9961 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa
    10021 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac
    10081 agttttattg ttcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
    10141 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
    10201 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctaac
    10261 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
    10321 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
    10381 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgg
//
