LOCUS       sequence-233127-       10533 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HF2-BE2 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10533)
  AUTHORS   Liang P, Sun H, Sun Y, Zhang X, Xie X, Zhang J, Zhang Z, Chen Y, 
            Ding C, Xiong Y, Ma W, Liu D, Huang J, Songyang Z
  TITLE     Effective gene editing by high-fidelity base editor 2 in mouse 
            zygotes.
  JOURNAL   Protein Cell. 2017 Aug;8(8):601-611. doi: 10.1007/s13238-017-0418-2.
            Epub 2017 Jun 5.
  PUBMED    28585179
REFERENCE   2  (bases 1 to 10533)
  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: 
            "10.1007/s13238-017-0418-2"; journalName: "Protein Cell"; date: 
            "2017-08"; volume: "8"; issue: "8"; pages: "601-611"
COMMENT     Sequence Label: pcDNA3.1(-)-HF2-BE2
FEATURES             Location/Qualifiers
     source          1..10533
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        7..386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..590
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     541..561
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     635..654
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        635..653
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             699..1385
                     /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="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
                     GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
                     SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
                     EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
                     LWATGLK"
     CDS             join(1434..2918,2919..2921,2922..3410,3411..3413,3414..3512,
                     3513..3515,3516..4205,4206..4208,4209..5534)
                     /codon_start=1
                     /product="Cas9 endonuclease from the Streptococcus pyogenes
                     Type II CRISPR/Cas system, mutated to improve targeting 
                     specificity (Kleinstiver et al., 2016)"
                     /label=SpCas9-HF2
                     /note="carries the mutations N497A, R661A, Q695A, Q926A, 
                     and D1135E"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTAFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGALSRKLINGIRDKQSGKTILDFLKSDGFANRNFMA
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRAITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFESPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5547..5795
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5808..5828
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(5896..5913)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5902..6126
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6172..6600
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6259..6278)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6469..6490
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6609..6629)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6614..6943
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6794..6929
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6856..6875
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             7010..7804
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7064..7083)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7674..7693
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7978..8099
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8015..8034)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8069..8088
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8148..8164)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8148..8164)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8161..8183)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8172..8188
                     /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(8196..8202,8203..8220,8221..8226))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8241..8262
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(8379..8396)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8550..9138)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8630..8649)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9309..10100,10101..10169))
                     /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     9932..9951
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10170..10274)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(10349..10368)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      241 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      301 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa
      601 ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc
      661 aagctggcta gcgtttaaac gggccctcta gagccaccat gagctcagag actggcccag
      721 tggctgtgga ccccacattg agacggcgga tcgagcccca tgagtttgag gtattcttcg
      781 atccgagaga gctccgcaag gagacctgcc tgctttacga aattaattgg gggggccggc
      841 actccatttg gcgacataca tcacagaaca ctaacaagca cgtcgaagtc aacttcatcg
      901 agaagttcac gacagaaaga tatttctgtc cgaacacaag gtgcagcatt acctggtttc
      961 tcagctggag cccatgcggc gaatgtagta gggccatcac tgaattcctg tcaaggtatc
     1021 cccacgtcac tctgtttatt tacatcgcaa ggctgtacca ccacgctgac ccccgcaatc
     1081 gacaaggcct gcgggatttg atctcttcag gtgtgactat ccaaattatg actgagcagg
     1141 agtcaggata ctgctggaga aactttgtga attatagccc gagtaatgaa gcccactggc
     1201 ctaggtatcc ccatctgtgg gtacgactgt acgttcttga actgtactgc atcatactgg
     1261 gcctgcctcc ttgtctcaac attctgagaa ggaagcagcc acagctgaca ttctttacca
     1321 tcgctcttca gtcttgtcat taccagcgac tgcccccaca cattctctgg gccaccgggt
     1381 tgaaaagcgg cagcgagact cccgggacct cagagtccgc cacacccgaa agtgataaaa
     1441 agtattctat tggtttagcc atcggcacta attccgttgg atgggctgtc ataaccgatg
     1501 aatacaaagt accttcaaag aaatttaagg tgttggggaa cacagaccgt cattcgatta
     1561 aaaagaatct tatcggtgcc ctcctattcg atagtggcga aacggcagag gcgactcgcc
     1621 tgaaacgaac cgctcggaga aggtatacac gtcgcaagaa ccgaatatgt tacttacaag
     1681 aaatttttag caatgagatg gccaaagttg acgattcttt ctttcaccgt ttggaagagt
     1741 ccttccttgt cgaagaggac aagaaacatg aacggcaccc catctttgga aacatagtag
     1801 atgaggtggc atatcatgaa aagtacccaa cgatttatca cctcagaaaa aagctagttg
     1861 actcaactga taaagcggac ctgaggttaa tctacttggc tcttgcccat atgataaagt
     1921 tccgtgggca ctttctcatt gagggtgatc taaatccgga caactcggat gtcgacaaac
     1981 tgttcatcca gttagtacaa acctataatc agttgtttga agagaaccct ataaatgcaa
     2041 gtggcgtgga tgcgaaggct attcttagcg cccgcctctc taaatcccga cggctagaaa
     2101 acctgatcgc acaattaccc ggagagaaga aaaatgggtt gttcggtaac cttatagcgc
     2161 tctcactagg cctgacacca aattttaagt cgaacttcga cttagctgaa gatgccaaat
     2221 tgcagcttag taaggacacg tacgatgacg atctcgacaa tctactggca caaattggag
     2281 atcagtatgc ggacttattt ttggctgcca aaaaccttag cgatgcaatc ctcctatctg
     2341 acatactgag agttaatact gagattacca aggcgccgtt atccgcttca atgatcaaaa
     2401 ggtacgatga acatcaccaa gacttgacac ttctcaaggc cctagtccgt cagcaactgc
     2461 ctgagaaata taaggaaata ttctttgatc agtcgaaaaa cgggtacgca ggttatattg
     2521 acggcggagc gagtcaagag gaattctaca agtttatcaa acccatatta gagaagatgg
     2581 atgggacgga agagttgctt gtaaaactca atcgcgaaga tctactgcga aagcagcgga
     2641 ctttcgacaa cggtagcatt ccacatcaaa tccacttagg cgaattgcat gctatactta
     2701 gaaggcagga ggatttttat ccgttcctca aagacaatcg tgaaaagatt gagaaaatcc
     2761 taacctttcg cataccttac tatgtgggac ccctggcccg agggaactct cggttcgcat
     2821 ggatgacaag aaagtccgaa gaaacgatta ctccctggaa ttttgaggaa gttgtcgata
     2881 aaggtgcgtc agctcaatcg ttcatcgaga ggatgaccgc ctttgacaag aatttaccga
     2941 acgaaaaagt attgcctaag cacagtttac tttacgagta tttcacagtg tacaatgaac
     3001 tcacgaaagt taagtatgtc actgagggca tgcgtaaacc cgcctttcta agcggagaac
     3061 agaagaaagc aatagtagat ctgttattca agaccaaccg caaagtgaca gttaagcaat
     3121 tgaaagagga ctactttaag aaaattgaat gcttcgattc tgtcgagatc tccggggtag
     3181 aagatcgatt taatgcgtca cttggtacgt atcatgacct cctaaagata attaaagata
     3241 aggacttcct ggataacgaa gagaatgaag atatcttaga agatatagtg ttgactctta
     3301 ccctctttga agatcgggaa atgattgagg aaagactaaa aacatacgct cacctgttcg
     3361 acgataaggt tatgaaacag ttaaagaggc gtcgctatac gggctgggga gccttgtcgc
     3421 ggaaacttat caacgggata agagacaagc aaagtggtaa aactattctc gattttctaa
     3481 agagcgacgg cttcgccaat aggaacttta tggccctgat ccatgatgac tctttaacct
     3541 tcaaagagga tatacaaaag gcacaggttt ccggacaagg ggactcattg cacgaacata
     3601 ttgcgaatct tgctggttcg ccagccatca aaaagggcat actccagaca gtcaaagtag
     3661 tggatgagct agttaaggtc atgggacgtc acaaaccgga aaacattgta atcgagatgg
     3721 cacgcgaaaa tcaaacgact cagaaggggc aaaaaaacag tcgagagcgg atgaagagaa
     3781 tagaagaggg tattaaagaa ctgggcagcc agatcttaaa ggagcatcct gtggaaaata
     3841 cccaattgca gaacgagaaa ctttacctct attacctaca aaatggaagg gacatgtatg
     3901 ttgatcagga actggacata aaccgtttat ctgattacga cgtcgatgcc attgtacccc
     3961 aatccttttt gaaggacgat tcaatcgaca ataaagtgct tacacgctcg gataagaacc
     4021 gagggaaaag tgacaatgtt ccaagcgagg aagtcgtaaa gaaaatgaag aactattggc
     4081 ggcagctcct aaatgcgaaa ctgataacgc aaagaaagtt cgataactta actaaagctg
     4141 agaggggtgg cttgtctgaa cttgacaagg ccggatttat taaacgtcag ctcgtggaaa
     4201 cccgcgccat cacaaagcat gttgcgcaga tactagattc ccgaatgaat acgaaatacg
     4261 acgagaacga taagctgatt cgggaagtca aagtaatcac tttaaagtca aaattggtgt
     4321 cggacttcag aaaggatttt caattctata aagttaggga gataaataac taccaccatg
     4381 cgcacgacgc ttatcttaat gccgtcgtag ggaccgcact cattaagaaa tacccgaagc
     4441 tagaaagtga gtttgtgtat ggtgattaca aagtttatga cgtccgtaag atgatcgcga
     4501 aaagcgaaca ggagataggc aaggctacag ccaaatactt cttttattct aacattatga
     4561 atttctttaa gacggaaatc actctggcaa acggagagat acgcaaacga cctttaattg
     4621 aaaccaatgg ggagacaggt gaaatcgtat gggataaggg ccgggacttc gcgacggtga
     4681 gaaaagtttt gtccatgccc caagtcaaca tagtaaagaa aactgaggtg cagaccggag
     4741 ggttttcaaa ggaatcgatt cttccaaaaa ggaatagtga taagctcatc gctcgtaaaa
     4801 aggactggga cccgaaaaag tacggtggct tcgagagccc tacagttgcc tattctgtcc
     4861 tagtagtggc aaaagttgag aagggaaaat ccaagaaact gaagtcagtc aaagaattat
     4921 tggggataac gattatggag cgctcgtctt ttgaaaagaa ccccatcgac ttccttgagg
     4981 cgaaaggtta caaggaagta aaaaaggatc tcataattaa actaccaaag tatagtctgt
     5041 ttgagttaga aaatggccga aaacggatgt tggctagcgc cggagagctt caaaagggga
     5101 acgaactcgc actaccgtct aaatacgtga atttcctgta tttagcgtcc cattacgaga
     5161 agttgaaagg ttcacctgaa gataacgaac agaagcaact ttttgttgag cagcacaaac
     5221 attatctcga cgaaatcata gagcaaattt cggaattcag taagagagtc atcctagctg
     5281 atgccaatct ggacaaagta ttaagcgcat acaacaagca cagggataaa cccatacgtg
     5341 agcaggcgga aaatattatc catttgttta ctcttaccaa cctcggcgct ccagccgcat
     5401 tcaagtattt tgacacaacg atagatcgca aacgatacac ttctaccaag gaggtgctag
     5461 acgcgacact gattcaccaa tccatcacgg gattatatga aactcggata gatttgtcac
     5521 agcttggggg tgactctggt ggttctacta atctgtcaga tattattgaa aaggagaccg
     5581 gtaagcaact ggttatccag gaatccatcc tcatgctccc agaggaggtg gaagaagtca
     5641 ttgggaacaa gccggaaagc gatatactcg tgcacaccgc ctacgacgag agcaccgacg
     5701 agaatgtcat gcttctgact agcgacgccc ctgaatacaa gccttgggct ctggtcatac
     5761 aggatagcaa cggtgagaac aagattaaga tgctctctgg tggttctccc aagaagaaga
     5821 ggaaagtcta attccaccac actggactag tggatccgag ctcggtacca agcttaagtt
     5881 taaaccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     5941 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     6001 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     6061 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     6121 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     6181 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     6241 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     6301 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     6361 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     6421 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     6481 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     6541 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     6601 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     6661 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     6721 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     6781 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     6841 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     6901 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     6961 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     7021 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     7081 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     7141 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     7201 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     7261 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     7321 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     7381 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     7441 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     7501 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     7561 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     7621 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     7681 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     7741 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     7801 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     7861 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     7921 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     7981 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     8041 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     8101 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     8161 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     8221 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     8281 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     8341 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     8401 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     8461 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     8521 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     8581 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     8641 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     8701 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     8761 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     8821 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     8881 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     8941 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     9001 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     9061 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     9121 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     9181 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     9241 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     9301 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     9361 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     9421 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     9481 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     9541 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     9601 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     9661 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     9721 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     9781 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     9841 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     9901 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     9961 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
    10021 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
    10081 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
    10141 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
    10201 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
    10261 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
    10321 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
    10381 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
    10441 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
    10501 ttgcgctgct tcgcgatgta cgggccagat ata
//
