LOCUS pSCBE2 13610 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses BE2 CRISPR base editor and gRNA scaffold in Streptomyces.
ACCESSION .
VERSION .
KEYWORDS pSCBE2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13610)
AUTHORS Zhong Z, Guo J, Deng L, Chen L, Wang J, Li S, Xu W, Deng Z, Sun Y
TITLE Base editing in Streptomyces with Cas9-deaminase fusions
JOURNAL BioRxiv 630137
REFERENCE 2 (bases 1 to 13610)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..13610
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(250..270)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(250..268)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(297..315)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 383..487
/gene="bla"
/label=AmpR promoter
CDS 488..1348
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(706..725)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 1519..2107
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2008..2027
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 2261..2278
/label=L4440
/note="L4440 vector, forward primer"
polyA_signal 2492..2626
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2516..2535)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2570..2589
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
promoter 5844..6145
/label=rpsL promoter
/note="promoter for the Xylanimonas cellulosilytica
ribosomal protein S12 gene"
CDS 6146..6832
/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 6881..10981
/codon_start=1
/product="catalytically dead mutant of the Cas9
endonuclease from the Streptococcus pyogenes Type II
CRISPR/Cas system"
/label=dCas9
/note="RNA-guided DNA-binding protein that lacks
endonuclease activity due to the D10A mutation in the RuvC
catalytic domain and the H840A mutation in the HNH
catalytic domain"
/translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 10994..11245
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
oriT 11608..11717
/note="incP origin of transfer"
CDS 11750..12121
/codon_start=1
/gene="traJ"
/product="oriT-recognizing protein"
/label=traJ
/translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
KQDELGKVMMGVVRPRAEP"
misc_RNA 12277..12352
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
CDS complement(12866..56)
/codon_start=1
/gene="aac(3)IV"
/product="aminoglycoside 3-N-acetyltransferase type IV
"
/label=ApmR
/note="confers resistance to apramycin (Paget and Davies,
1996)"
/translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
ORIGIN
1 agctcggctt ttcgccattc gtattgcacg acattgcact ccaccgctga tgacatcagt
61 cgatcatagc acgatcaacg gcactgttgc aaatagtcgg tggtgataaa cttatcatcc
121 ccttttgctg atggagctgc acatgaaccc attcaaaggc cggcattttc agcgtgacat
181 cattctgtgg gccgtacgct ggtactgcaa atacggcatc agttaccgtg agctgcaggt
241 cgacggatcc ctttagtgag ggttaattgc ggccgcgaat tcttgaagac gaaagggcct
301 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
361 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
421 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
481 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
541 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
601 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
661 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
721 attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
781 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
841 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
901 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
961 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
1021 cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
1081 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
1141 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
1201 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
1261 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
1321 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
1381 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
1441 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
1501 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
1561 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
1621 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
1681 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
1741 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
1801 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
1861 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
1921 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
1981 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
2041 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
2101 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
2161 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
2221 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
2281 agcggaagag cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca
2341 ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc
2401 gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca
2461 acgacaggag cacgatcatg cgcacccgtc agatccagac atgataagat acattgatga
2521 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
2581 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttcccg cacgatcagc
2641 gtcccgccga ccaggagcag tgcggccagc actgccgcta acgcctggtc ctggtcccgg
2701 tcctggtggt gcatcagtcc tccccgtgat cacttcggca cccaccgtag tgatcacccc
2761 cgacagcgga tcaaggggtt tgcgggtccc ggtcggcgcc gggcggggga ggcaggagcc
2821 gccgacgctg cctctgggac gggccggacg gcagggggac cggcggccgg gcgagctgca
2881 gccgggggtc cggcagggcc ggagcgggcg gaaccgtgct ctgacctgcg gcccgagttt
2941 cgtcacgtga cggaatggaa ggctgctgca tttcgtcacg tgacgtatct cggcgagcga
3001 ctgccgacgc cacggcggac acgatcgcct cgcgctggcg ccgggcctcg tacgcccgct
3061 ggcggcagga gcggcggcag tagtcccggc tccggccgac gccggattgc ttgatctccg
3121 agccgcacca ggcgcagagc ttcgcgccgt cggcgtccct gggggtggtg gtgctcatgg
3181 ccgacgaccg tacgcggcac gtctcgtagc gaggcgagtc gggcgcgagg taccgcctgc
3241 acgaagtgcc ggcggggccg accccgggcg agtaatccca ggattactcc cgcggcttcg
3301 accccggccg ccgtcgccgc gtacgtcacc gacccccgcc gtacgtcacc gggatgacgt
3361 acggcggggg ggagcgagtt agtgcgaagt gggcccactt gcgagccggg cgatgtgccg
3421 ggcggcccgc tcctggcggt cgtcggcgtc gtcgtcctgg tcgtcgtcct gctctcgccg
3481 tcggcgtgca gttgcttcct cgcggcgctg ggcgagggcg gcgagcatgt cggcgtacgc
3541 ctcggccacc tcccccgccg tgagcaccac cactgtgtcg gccgcgtcgg ccagcgccag
3601 gacctcccgc acccgttcgc ccacggccgc cgaatcctcg ttgccgtcct tgccttcggc
3661 ggcccgggtc gcctcgaggt cgagggcgcg gcgggtgacc gcgtgccatc cgtcctcggt
3721 cacggcgacc ccggcccgca gctccccgcc gtcggcgtcg gccgccagga gcagatcgag
3781 gtcgtcggcc tcggtgtcgc cgccgtcgag cccgagcatc tgccgcaggt agcgggtcca
3841 ttcgatggcc cggcgtcccc gggttgcccg ctcgtactcg tgccagcgcg agaggttcca
3901 ctccagcgag ccgaccccgg cggcgtcgtc ctcggtcatg ccgccggtca ggtccccgat
3961 ccgtccgagg agttcgaacg gggcgacgtt cccgccggtc gccgtcttga ggtcggcgcg
4021 ggcgagttcg agggcgggcg ccttcccgtc ctgggtcttg gcgatgtact cggcgaggtc
4081 gttggcgtcg cgctcggtct ccagccgctt gaagtcgacg ccgtgccggt cgtcgggcgt
4141 gaaggcgggg ttgaccttgc gcagggcggc ggtccacacg gaccgccagt gcccctgcca
4201 ctcgtcgagc gcggcgccgg tcggctcgaa ggtggcgacg atctgcttcg cggaccgctc
4261 cccctcggtc cggccgccga ccaggacgat cgcgtggatg tgcgggtgcc agccgttgat
4321 ctgccccacg gtgacttcgg tcgcgcggat catgccgacg tacccgatcc ggtctcggat
4381 gccctcgcgg tcggcggccc ggtgcccgtc cttggcccgg cgtccggccc acgtgccgcc
4441 cgtgatcagt cgctggtagg cgcccggccg ccgggggctg tccggcgtct tccgggtgcc
4501 ctggagggcg tccatgaggt ccgcgagccg gtccgtgtgc ccatggcggg ccgtgaaggt
4561 gaccaggtag gcggtccccc cgcgcttgat ccactcgacc acggcggcgg tgatctcctc
4621 ggcccgcttg tgccggatcg tggcggcgca gaccgggcag agccagatcc gcccgcaccg
4681 catcaggccc aggaccacgg acgttccggc cgccgtctgg gcgacgatca cgccggaggc
4741 agggtccatc agggcgcggc cgcagccctt gcacgcggcg tccccgctga tccgccacag
4801 cgtccggcgg cggctgtacc gggcggcttt ccgcagtcgg gcagcgtcgc tccgcgacgt
4861 gcttcctact tccgagaggc tgtcgcctct cgggctctcc ccatccaccc cgtccggaga
4921 aaccgcaggt cggaggggtg cgggaaactc tgttgtttct ttcccaaggt gttcgctttt
4981 gcctcgggcg gcatctcgcg tcacacgcgc gatcgcccgc ttcgctgcca tccggcagcg
5041 gtctgagcag tagatacgcg gccgtttgcc cggtgtgtgg gcaattgcgg tcccgcagtg
5101 gcagcggggc ccggcgggcc gatctggcaa tgcctcggca tcgctccgta ctctgggcac
5161 gagcaacgtt cctgtctcgc ccggctaagg ggcgcgagtc tgggagcgga cgggtcggag
5221 gtgcgaagtc cggcccgttg ctctttggtc tggtgggaat cctggcacca atcgggccag
5281 aggttccctc cgccactccc gacgcccctt ggggctggtg tgacttggag ggccgaagag
5341 agccccgccg gtgatccggc ggggctttga cgtgcggtca gtgcgtgtgt cggcgagcga
5401 tggccacgag gccctggaag ccgagcggtc cggcgaagtc ggcccagtcg caaccgggct
5461 cagcgcagtg ggcggaccag ccaccgccgt tggggtcctg gaccaggttc acggtcccct
5521 cggtcaggcg tccgtcgaag tcggtcatgg tcggtctcct ggtgggtggg ggcggggcgc
5581 cagcacgaag tgccggcgcc ccgcgggggt tggtcgggtc aggcgccgaa ccggcgggcg
5641 gcggcggcga ccaggccgtc ggcggcggcc atggcgcggt cgcggtcggt ggtgagggcg
5701 gtgcggtcgg cggcggccca gtcctgctct cacgcaacgt ctacgtggac gctcagggcg
5761 acacgatcga ggtcggggag tccgtcgctc cggcgggccg ctggcgcgtc caccgggact
5821 gatcaaggcg aatacttcat atggccctgc aggcggaagt caggtagaca cgacttccgc
5881 tagtccttgc aaggtctgct gacgtgaggc ggggcggtcg tttttgaccg ccctgccttc
5941 gtcatgtagg ctcgctcgct gtgcctggcg tgtcatcaga cgcccaggtc ccggtgccgt
6001 gaggcccggg ccatcgagcc ggtggtacgt ggctgcggtc cccttgtgag ggctgcgcgc
6061 cgtgtgctgt ccggcgcgca cagccttgaa tccacccgcg ggggccggcc ggtctccgtg
6121 agctcgagta gacgacggag acgtaatgtc gtccgagacc ggcccggtgg ccgtggaccc
6181 gaccctgcgt cgccgcatcg agccgcacga gttcgaggtg ttcttcgacc cgcgggagct
6241 gcgcaaggag acctgcctgc tgtacgagat caactggggc ggccggcact cgatctggcg
6301 ccacacctcg cagaacacca acaagcacgt cgaggtgaac ttcatcgaga agttcaccac
6361 cgagcggtac ttctgcccga acacccgctg ctcgatcacc tggttcctgt cctggtcccc
6421 gtgcggcgag tgctcccggg cgatcaccga gttcctgtcc cgctacccgc acgtcaccct
6481 gttcatctac atcgcccggc tgtaccacca cgccgacccg cggaaccggc agggcctgcg
6541 cgacctgatc tcctccggcg tgaccatcca gatcatgacc gagcaggagt ccggctactg
6601 ctggcggaac ttcgtcaact actcgccgtc caacgaggcc cactggccgc ggtacccgca
6661 cctgtgggtg cgcctgtacg tcctggagct gtactgcatc atcctgggcc tgccgccgtg
6721 cctgaacatc ctgcgtcgca agcagccgca gctgaccttc ttcaccatcg ccctgcagtc
6781 ctgccactac cagcgcctgc cgccgcacat cctgtgggcg accggcctga agtccggctc
6841 ggagaccccg ggcacctcgg agtccgccac cccggagtcc gacaagaagt acagcatcgg
6901 cctggccatc ggcaccaaca gcgtgggctg ggcggtcatc accgacgagt acaaggtccc
6961 ctccaagaag ttcaaggtcc tgggcaacac cgaccggcac tcgatcaaga agaacctgat
7021 cggcgccctg ctcttcgaca gcggcgagac cgccgaggcg acccgcctga agcggaccgc
7081 gcgccggcgc tacacccggc gcaagaaccg catctgctac ctccaggaaa tcttctccaa
7141 cgagatggcc aaggtcgacg actcgttctt ccaccggctc gaggagagct tcctggtgga
7201 ggaggacaag aagcacgagc gccacccgat cttcggcaac atcgtcgacg aggtggccta
7261 ccacgagaag taccccacca tctaccacct ccgcaagaag ctggtcgact cgaccgacaa
7321 ggcggacctg cggctcatct acctggccct cgcgcacatg atcaagttcc gcggccactt
7381 cctcatcgag ggcgacctga acccggacaa ctccgacgtc gacaagctct tcatccagct
7441 ggtgcagacc tacaaccagc tgttcgagga gaaccccatc aacgccagcg gcgtcgacgc
7501 caaggcgatc ctctccgcgc gcctgagcaa gtcccggcgc ctggagaacc tcatcgccca
7561 gctgccgggc gagaagaaga acggcctctt cggcaacctg atcgcgctgt cgctcggcct
7621 gacccccaac ttcaagagca acttcgacct ggccgaggac gcgaagctcc agctgtccaa
7681 ggacacctac gacgacgacc tggacaacct gctcgcccag atcggcgacc agtacgcgga
7741 cctcttcctg gccgcgaaga acctctcgga cgccatcctg ctcagcgaca tcctgcgggt
7801 caacaccgag atcaccaagg ccccgctgtc ggcgagcatg atcaagcggt acgacgagca
7861 ccaccaggac ctgaccctgc tcaaggccct cgtgcgccag cagctgcccg agaagtacaa
7921 ggaaatcttc ttcgaccagt ccaagaacgg ctacgccggc tacatcgacg gcggcgcgtc
7981 gcaggaggag ttctacaagt tcatcaagcc gatcctggag aagatggacg gcaccgagga
8041 gctgctcgtc aagctgaacc gcgaggacct gctccgcaag cagcggacct tcgacaacgg
8101 ctccatcccg caccagatcc acctgggcga gctgcacgcc atcctccggc gccaggagga
8161 cttctacccc ttcctgaagg acaaccgcga gaagatcgag aagatcctga ccttccgcat
8221 cccgtactac gtcggccccc tggcccgcgg caactcccgg ttcgcgtgga tgacccggaa
8281 gtcggaggag accatcaccc cgtggaactt cgaggaggtc gtggacaagg gcgcctccgc
8341 ccagtcgttc atcgagcgca tgaccaactt cgacaagaac ctcccgaacg agaaggtcct
8401 gcccaagcac tccctgctct acgagtactt caccgtgtac aacgagctga ccaaggtcaa
8461 gtacgtgacc gagggcatgc ggaagccggc cttcctgtcg ggcgagcaga agaaggcgat
8521 cgtcgacctg ctcttcaaga ccaaccgcaa ggtcaccgtg aagcagctga aggaggacta
8581 cttcaagaag atcgagtgct tcgactccgt cgagatcagc ggcgtggagg accgcttcaa
8641 cgcctccctg ggcacctacc acgacctgct caagatcatc aaggacaagg acttcctcga
8701 caacgaggag aacgaggaca tcctggagga catcgtcctc accctgaccc tcttcgagga
8761 ccgcgagatg atcgaggagc ggctcaagac ctacgcccac ctgttcgacg acaaggtgat
8821 gaagcagctg aagcggcgcc ggtacaccgg ctggggccgc ctctcccgga agctgatcaa
8881 cggcatccgg gacaagcaga gcggcaagac catcctggac ttcctcaagt ccgacggctt
8941 cgccaaccgc aacttcatgc agctcatcca cgacgacagc ctgaccttca aggaggacat
9001 ccagaaggcc caggtctcgg gccagggcga cagcctccac gagcacatcg ccaacctggc
9061 gggctccccg gcgatcaaga agggcatcct ccagaccgtc aaggtcgtgg acgagctggt
9121 caaggtgatg ggccgccaca agcccgagaa catcgtgatc gagatggccc gggagaacca
9181 gaccacccag aagggccaga agaactcgcg cgagcggatg aagcggatcg aggagggcat
9241 caaggagctg ggcagccaga tcctgaagga gcacccggtc gagaacaccc agctccagaa
9301 cgagaagctg tacctctact acctccagaa cggccgcgac atgtacgtgg accaggagct
9361 ggacatcaac cggctgtccg actacgacgt cgacgccatc gtgccgcagt ccttcctgaa
9421 ggacgactcg atcgacaaca aggtcctgac ccgctcggac aagaaccggg gcaagtccga
9481 caacgtgccc tcggaggagg tcgtgaagaa gatgaagaac tactggcgcc agctgctcaa
9541 cgccaagctc atcacccagc gcaagttcga caacctgacc aaggccgagc ggggcggcct
9601 gagcgagctg gacaaggcgg gcttcatcaa gcgccagctg gtcgagaccc ggcagatcac
9661 caagcacgtg gcccagatcc tggactcccg gatgaacacc aagtacgacg agaacgacaa
9721 gctgatccgc gaggtcaagg tgatcaccct caagagcaag ctggtctccg acttccgcaa
9781 ggacttccag ttctacaagg tccgggagat caacaactac caccacgccc acgacgcgta
9841 cctgaacgcc gtcgtgggca ccgcgctgat caagaagtac ccgaagctgg agtccgagtt
9901 cgtctacggc gactacaagg tctacgacgt gcgcaagatg atcgccaaga gcgagcagga
9961 gatcggcaag gccaccgcga agtacttctt ctactccaac atcatgaact tcttcaagac
10021 cgagatcacc ctggccaacg gcgagatccg caagcggccc ctgatcgaga ccaacggcga
10081 gaccggcgag atcgtctggg acaagggccg cgacttcgcc accgtccgga aggtgctgtc
10141 gatgccgcag gtcaacatcg tgaagaagac cgaggtgcag accggcggct tcagcaagga
10201 gtccatcctc cccaagcgca acagcgacaa gctgatcgcc cggaagaagg actgggaccc
10261 gaagaagtac ggcggcttcg acagccccac cgtcgcctac tccgtgctgg tcgtggcgaa
10321 ggtcgagaag ggcaagagca agaagctgaa gtccgtgaag gagctgctcg gcatcaccat
10381 catggagcgc tcctcgttcg agaagaaccc gatcgacttc ctggaggcca agggctacaa
10441 ggaggtcaag aaggacctca tcatcaagct gcccaagtac agcctgttcg agctggagaa
10501 cggccgcaag cggatgctcg cctccgcggg cgagctgcaa aagggcaacg agctggccct
10561 cccgtcgaag tacgtcaact tcctgtacct cgcgtcccac tacgagaagc tgaagggctc
10621 gcccgaggac aacgagcaga agcagctctt cgtggagcag cacaagcact acctggacga
10681 gatcatcgag cagatcagcg agttcagcaa gcgcgtcatc ctggccgacg cgaacctcga
10741 caaggtgctg tccgcctaca acaagcaccg cgacaagccg atccgggagc aggcggagaa
10801 catcatccac ctgttcaccc tcaccaacct gggtgccccg gccgccttca agtacttcga
10861 caccaccatc gaccgcaagc ggtacacctc caccaaggag gtcctcgacg cgaccctgat
10921 ccaccagagc atcaccggcc tgtacgagac ccgcatcgac ctgagccagc tgggcggcga
10981 ctcgggcggc tccaccaacc tgtcggacat catcgagaag gagaccggca agcagctggt
11041 gatccaggag tccatcctga tgctgccgga ggaagtggag gaagtgatcg gcaacaagcc
11101 ggagtcggac atcctggtgc acaccgccta cgacgagtcg accgacgaga acgtcatgct
11161 gctgacctcg gacgccccgg agtacaagcc gtgggccctg gtcatccagg actccaacgg
11221 cgagaacaag atcaagatgc tgtgagaatt cagatctacg cgttccccgc aaaagcggcc
11281 tttgactccc tgcaagcctc agcgaccgaa tatatcggtt atgcgtgggc gatggttgtt
11341 gtcattgtcg gcgcaactat cggtatcaag ctgtttaaga aattcacctc gaaagcaagc
11401 tgataaaccg atacaattaa aggctccttt tggagccttt ttttaagctt gcatgcctgc
11461 aggtcgacgg atcttttccg ctgcataacc ctgcttcggg gtcattatag cgattttttc
11521 ggtatatcca tcctttttcg cacgatatac aggattttgc caaagggttc gtgtagactt
11581 tccttggtgt atccaacggc gtcagccggg caggataggt gaagtaggcc cacccgcgag
11641 cgggtgttcc ttcttcactg tcccttattc gcacctggcg gtgctcaacg ggaatcctgc
11701 tctgcgaggc tggccggcta ccgccggcgt aacagatgag ggcaagcgga tggctgatga
11761 aaccaagcca accaggaagg gcagcccacc tatcaaggtg tactgccttc cagacgaacg
11821 aagagcgatt gaggaaaagg cggcggcggc cggcatgagc ctgtcggcct acctgctggc
11881 cgtcggccag ggctacaaaa tcacgggcgt cgtggactat gagcacgtcc gcgagctggc
11941 ccgcatcaat ggcgacctgg gccgcctggg cggcctgctg aaactctggc tcaccgacga
12001 cccgcgcacg gcgcggttcg gtgatgccac gatcctcgcc ctgctggcga agatcgaaga
12061 gaagcaggac gagcttggca aggtcatgat gggcgtggtc cgcccgaggg cagagccatg
12121 acttttttag ccgctaaaac ggccgggggg tgcgcgtgat tgccaagcac gtccccatgc
12181 gctccatcaa gaagagcgac ttcgcggagc tggtgaagta catcaccgac gagcaaggca
12241 agaccgatcc ccggggacct gcaggtcgac tctagagttt tagagctaga aatagcaagt
12301 taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gctaaaaaaa
12361 aaccccgccc ctgacagggc ggggtttttt ttgtagcctc aggagttgta gctgttcaca
12421 ttcgaacggt ctctgctttg acaacatgct gtgcggtgtt gtaaagtcgt ggccgctagc
12481 tatttacccg caggacatat ccacgccctc ctacatcgaa gctgaaagca cgagattctt
12541 cgccctccga gagctgcatc aggtcggaga cgctgtcgaa cttttcgatc agaaacttct
12601 cgacagacgt cgcggtgagt tcaggctttt tcatatctca ttgcccccgg acgagcgtct
12661 gctccgccat tcgccgtccg ccgtgccaat cggatcagcc gtccaaatgc gggattttcg
12721 ttagtcggag gccaaacggc attgagcgtc agcatatcat cagcgagctg aagaaagaca
12781 atccccgatc cgctccacgt gttgccccag caatcagcgc gaccttgccc ctccaacgtc
12841 atctcgttct ccgctcatga gctcagccaa tcgactggcg agcggcatcg cattcttcgc
12901 atcccgcctc tggcggatgc aggaagatca acggatctcg gcccagttga cccagggctg
12961 tcgccacaat gtcgcgggag cggatcaacc gagcaaaggc atgaccgact ggaccttcct
13021 tctgaaggct cttctccttg agccacctgt ccgccaaggc aaagcgctca cagcagtggt
13081 cattctcgag ataatcgacg cgtaccaact tgccatcctg aagaatggtg cagtgtctcg
13141 gcaccccata gggaaccttt gccatcaact cggcaagatg cagcgtcgtg ttggcatcgt
13201 gtcccacgcc gaggagaagt acctgcccat cgagttcatg gacacgggcg accgggcttg
13261 caggcgagtg aggtggcagg ggcaatggat cagagatgat ctgctctgcc tgtggccccg
13321 ctgccgcaaa ggcaaatgga tgggcgctgc gctttacatt tggcaggcgc cagaatgtgt
13381 cagagacaac tccaaggtcc ggtgtaacgg gcgacgtggc aggatcgaac ggctcgtcgt
13441 ccagacctga ccacgagggc atgacgagcg tccctcccgg acccagcgca gcacgcaggg
13501 cctcgatcag tccaagtggc ccatcttcga ggggccggac gctacggaag gagctgtgga
13561 ccagcagcac accgccgggg gtaaccccaa ggttgagaag ctgaccgatg
//
