epi-BE4max

LOCUS       epi-BE4max      14566 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses AncBE4max, EBNA1 and blasticidin resistence gene; cloning 
            backbone for sgRNA; Contains Epstein-Barr virus oriP replication 
            origin .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14566)
  AUTHORS   Qi T, Wu F, Xie Y, Gao S, Li M, Pu J, Li D, Lan F, Wang Y
  TITLE     Base Editing Mediated Generation of Point Mutations Into Human 
            Pluripotent Stem Cells for Modeling Disease.
  JOURNAL   Front Cell Dev Biol. 2020 Sep 25;8:590581. doi: 
            10.3389/fcell.2020.590581. eCollection 2020.
  PUBMED    33102492
REFERENCE   2  (bases 1 to 14566)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Front Cell 
            Dev Biol."; date: "2020-09-25"; volume: "8:590581. doi"; pages: " 
            10.3389/fcell.2020.590581. eCollection 2020"
COMMENT     Sequence Label: epi-BE4max
FEATURES             Location/Qualifiers
     source          1..14566
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(880..898)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteAam of EcoRI site, forward 
                     primer"
     promoter        966..1070
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1071..1139,1140..1931)
                     /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(1289..1308)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2102..2690
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2591..2610
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2960..3273
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     3192..3212
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     misc_RNA        3294..3369
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    3417..3534
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3603..3814
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             3876..3896
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(4677..4700,4701..4703,4704..8777)
                     /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             4677..8777
                     /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             8808..9056
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             9087..9335
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             9378..9398
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9407..9424
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(9447..9464)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    9453..9677
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        9725..10054
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      9905..10040
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             10076..10474
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    10640..10774
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10677..10696)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10731..10750
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      11414..13203
                     /label=oriP
                     /note="Epstein-Barr virus oriP replication origin (Yates et
                     al., 2000)"
ORIGIN
        1 cmcgtcctct accccggcgg cctccactac ctcctcgacc ccggcctcca ctacctcctc
       61 gaccccggcc tccactgcct cctcgacccc ggcctccacc tcctgctcct gcccctcctg
      121 ctcctgcccc tcctcctgct cctgcccctc ctgcccctcc tgctcctgct cctgctcctg
      181 ttccaccgtg ggtccctttg cagccaatgc aacttggacg tttttggggt ctccggacac
      241 catctctatg tcttggccct gatcctgagc cgcccggggc tcctggtctt ccgcctcctc
      301 gtcctcgtcc tcttccccgt cctcgtccat ggttatcacc ccctcttctt tgaggtccac
      361 tgccgccgga gccttctggt ccagatgtgt ctcccttctc tcctaggcca tttccaggtc
      421 ctgtacctgg cccctcgtca gacatgattc acactaaaag agatcaatag acatctttat
      481 tagacgacgc tcagtgaata cagggagtgc agactcctgc cccctccaac agccccccca
      541 ccctcatccc cttcatggtc gctgtcagac agatccaggt ctgaaaattc cccatcctcc
      601 gaaccatcct cgtcctcatc accaattact cgcagcccgg aaaactcccg ctgaacatcc
      661 tcaagatttg cgtcctgagc ctcaagccag gcctcaaatt cctcgtcccc ctttttgctg
      721 gacggtaggg atggggattc tcgggacccc tcctcttcct cttcaaggtc accagacaga
      781 gatgctactg gggcaacgga agaaaagctg ggtgcggcct gtgaggatca gcttatcgat
      841 gataagctgt caaacatgag aattcttgaa gacgaaaggg cctcgtgata cgcctatttt
      901 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
      961 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     1021 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     1081 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     1141 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     1201 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     1261 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
     1321 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     1381 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     1441 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     1501 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     1561 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
     1621 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     1681 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     1741 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     1801 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     1861 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     1921 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     1981 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     2041 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     2101 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     2161 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     2221 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     2281 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     2341 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     2401 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     2461 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     2521 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     2581 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     2641 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     2701 acgcggcctt tttacggttc ctggcctttt gctggccttg aagctgtccc tgatggtcgt
     2761 catctacctg cctggacagc atggcctgca acgcgggcat cccgatgccg ccggaagcga
     2821 gaagaatcat aatggggaag gccatccagc ctcgcgtcga actagatgat ccggctgtgg
     2881 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2941 agcatgcatt atgtctagag atccgacgcc gccatctcta ggcccgcgcc ggccccctcg
     3001 cacagacttg tgggagaagc tcggctactc ccctgccccg gttaatttgc atataatatt
     3061 tcctagtaac tatagaggct taatgtgcga taaaagacag ataatctgtt ctttttaata
     3121 ctagctacat tttacatgat aggcttggat ttctataaga gatacaaata ctaaattatt
     3181 attttaaaaa acagcacaaa aggaaactca ccctaactgt aaagtaattg tgtgttttga
     3241 gactataaat atgcatgcga gaaaagcctt gtttagaaga gcgatgctct tcggttttag
     3301 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg
     3361 agtcggtgct tttttgaatt ctagatcttg agacaaatgg cagtattcat ccacaatttt
     3421 aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca
     3481 acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt
     3541 tattacaggg acagcagaga tccactttgg cgccggctcg agtggctccg gtgcccgtca
     3601 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg
     3661 aaccggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct
     3721 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt
     3781 tctttttcgc aacgggtttg ccgccagaac acaggtgtcg tgacgcggga tccgccacca
     3841 tgaaacggac agccgacgga agcgagttcg agtcaccaaa gaagaagcgg aaagtcagca
     3901 gtgaaaccgg accagtggca gtggacccaa ccctgaggag acggattgag ccccatgaat
     3961 ttgaagtgtt ctttgaccca agggagctga ggaaggagac atgcctgctg tacgagatca
     4021 agtggggcac aagccacaag atctggcgcc acagctccaa gaacaccaca aagcacgtgg
     4081 aagtgaattt catcgagaag tttacctccg agcggcactt ctgcccctct accagctgtt
     4141 ccatcacatg gtttctgtct tggagccctt gcggcgagtg ttccaaggcc atcaccgagt
     4201 tcctgtctca gcaccctaac gtgaccctgg tcatctacgt ggcccggctg tatcaccaca
     4261 tggaccagca gaacaggcag ggcctgcgcg atctggtgaa ttctggcgtg accatccaga
     4321 tcatgacagc cccagagtac gactattgct ggcggaactt cgtgaattat ccacctggca
     4381 aggaggcaca ctggccaaga tacccacccc tgtggatgaa gctgtatgca ctggagctgc
     4441 acgcaggaat cctgggcctg cctccatgtc tgaatatcct gcggagaaag cagccccagc
     4501 tgacattttt caccattgct ctgcagtctt gtcactatca gcggctgcct cctcatattc
     4561 tgtgggctac aggcctgaag tctggaggat ctagcggagg atcctctggc agcgagacac
     4621 caggaacaag cgagtcagca acaccagaga gcagtggcgg cagcagcggc ggcagcgaca
     4681 agaagtacag catcggcctg gccatcggca ccaactctgt gggctgggcc gtgatcaccg
     4741 acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca
     4801 tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc
     4861 ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc
     4921 aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag
     4981 agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg
     5041 tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg
     5101 tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca
     5161 agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca
     5221 agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg
     5281 ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg
     5341 aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg
     5401 ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca
     5461 aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg
     5521 gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga
     5581 gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca
     5641 agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc
     5701 tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca
     5761 ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga
     5821 tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc
     5881 ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg cacgccattc
     5941 tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga
     6001 tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg
     6061 cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg
     6121 acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc
     6181 ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg
     6241 agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg
     6301 agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc
     6361 agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg
     6421 tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg
     6481 acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc
     6541 tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt
     6601 tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga
     6661 gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc
     6721 tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga
     6781 cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc
     6841 acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg
     6901 tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa
     6961 tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc
     7021 ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa
     7081 acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt
     7141 acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc
     7201 ctcagagctt tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga
     7261 accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact
     7321 ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg
     7381 ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg
     7441 aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt
     7501 acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg
     7561 tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc
     7621 acgcccacga cgcctaccta aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta
     7681 agctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg
     7741 ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca
     7801 tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga
     7861 tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg
     7921 tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag
     7981 gcggcttcag caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa
     8041 agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg
     8101 tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc
     8161 tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg
     8221 aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc
     8281 tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg
     8341 gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg
     8401 agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca
     8461 agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg
     8521 ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca
     8581 gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg
     8641 ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc
     8701 tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt
     8761 ctcagctggg aggtgacagc ggcgggagcg gcgggagcgg ggggagcact aatctgagcg
     8821 acatcattga gaaggagact gggaaacagc tggtcattca ggagtccatc ctgatgctgc
     8881 ctgaggaggt ggaggaagtg atcggcaaca agccagagtc tgacatcctg gtgcacaccg
     8941 cctacgacga gtccacagat gagaatgtga tgctgctgac ctctgacgcc cccgagtata
     9001 agccttgggc cctggtcatc caggattcta acggcgagaa taagatcaag atgctgagcg
     9061 gaggatccgg aggatctgga ggcagcacca acctgtctga catcatcgag aaggagacag
     9121 gcaagcagct ggtcatccag gagagcatcc tgatgctgcc cgaagaagtc gaagaagtga
     9181 tcggaaacaa gcctgagagc gatatcctgg tccataccgc ctacgacgag agtaccgacg
     9241 aaaatgtgat gctgctgaca tccgacgccc cagagtataa gccctgggct ctggtcatcc
     9301 aggattccaa cggagagaac aaaatcaaaa tgctgtctgg cggctcaaaa agaaccgccg
     9361 acggcagcga attcgagccc aagaagaaga ggaaagtcta accggtcatc atcaccatca
     9421 ccattgagtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     9481 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     9541 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     9601 tggggtgggg caggacagca agggggagga ttgggaagag aatagcaggc atgctgggga
     9661 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcga taccgctgtg
     9721 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     9781 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     9841 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     9901 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     9961 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
    10021 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt ggataatggc
    10081 caagcctttg tctcaagaag aatccaccct cattgaaaga gcaacggcta caatcaacag
    10141 catccccatc tctgaagact acagcgtcgc cagcgcagct ctctctagcg acggccgcat
    10201 cttcactggt gtcaatgtat atcattttac tgggggacct tgtgcagaac tcgtggtgct
    10261 gggcactgct gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga tcggaaatga
    10321 gaacaggggc atcttgagcc cctgcggacg gtgccgacag gtgcttctcg atctgcatcc
    10381 tgggatcaaa gccatagtga aggacagtga tggacagccg acggcagttg ggattcgtga
    10441 attgctgccc tctggttatg tgtgggaggg ctaagaattc ttcgaaatga ccgaccaagc
    10501 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
    10561 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
    10621 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
    10681 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
    10741 caaactcatc aatgtatctt atcatgtctg gatcccttga aatacatcca caggtacacc
    10801 tccaattgac tcaaatgatg tcaattagtc tatcagaagc ttgtggaagg ctggagacgg
    10861 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
    10921 gtgttggcgg gtgtcggggc gcagccatga ggtcgactct agaggatcga tccccgcccc
    10981 ggacgaacta aacctgacta cgacatctct gccccttctt cgcggggcag tgcatgtaat
    11041 cccttcagtt ggttggtaca acttgccaac tgggccctgt tccacatgtg acacgggggg
    11101 ggaccaaaca caaaggggtt ctctgactgt agttgacatc cttataaatg gatgtgcaca
    11161 tttgccaaca ctgagtggct ttcatcctgg agcagacttt gcagtctgtg gactgcaaca
    11221 caacattgcc tttatgtgta actcttggct gaagctctta caccaatgct gggggacatg
    11281 tacctcccag gggcccagga agactacggg aggctacacc aacgtcaatc agaggggcct
    11341 gtgtagctac cgataagcgg accctcaaga gggcattagc aatagtgttt ataaggcccc
    11401 cttgttaacc ctaaacgggt agcatatgct tcccgggtag tagtatatac tatccagact
    11461 aaccctaatt caatagcata tgttacccaa cgggaagcat atgctatcga attagggtta
    11521 gtaaaagggt cctaaggaac agcgatatct cccaccccat gagctgtcac ggttttattt
    11581 acatggggtc aggattccac gagggtagtg aaccatttta gtcacaaggg cagtggctga
    11641 agatcaagga gcgggcagtg aactctcctg aatcttcgcc tgcttcttca ttctccttcg
    11701 tttagctaat agaataactg ctgagttgtg aacagtaagg tgtatgtgag gtgctcgaaa
    11761 acaaggtttc aggtgacgcc cccagaataa aatttggacg gggggttcag tggtggcatt
    11821 gtgctatgac accaatataa ccctcacaaa ccccttgggc aataaatact agtgtaggaa
    11881 tgaaacattc tgaatatctt taacaataga aatccatggg gtggggacaa gccgtaaaga
    11941 ctggatgtcc atctcacacg aatttatggc tatgggcaac acataatcct agtgcaatat
    12001 gatactgggg ttattaagat gtgtcccagg cagggaccaa gacaggtgaa ccatgttgtt
    12061 acactctatt tgtaacaagg ggaaagagag tggacgccga cagcagcgga ctccactggt
    12121 tgtctctaac acccccgaaa attaaacggg gctccacgcc aatggggccc ataaacaaag
    12181 acaagtggcc actctttttt ttgaaattgt ggagtggggg cacgcgtcag cccccacacg
    12241 ccgccctgcg gttttggact gtaaaataag ggtgtaataa cttggctgat tgtaaccccg
    12301 ctaaccactg cggtcaaacc acttgcccac aaaaccacta atggcacccc ggggaatacc
    12361 tgcataagta ggtgggcggg ccaagatagg ggcgcgattg ctgcgatctg gaggacaaat
    12421 tacacacact tgcgcctgag cgccaagcac agggttgttg gtcctcatat tcacgaggtc
    12481 gctgagagca cggtgggcta atgttgccat gggtagcata tactacccaa atatctggat
    12541 agcatatgct atcctaatct atatctgggt agcataggct atcctaatct atatctgggt
    12601 agtatatgct atcctaatct gtatccgggt agcatatgct atcctaatag agattagggt
    12661 agcatatgct atcctaatct atatctgggt agcataggct atcctaatct atatctgggt
    12721 agtatatgct atcctaatct gtatccgggt agcatatgct atcctaatag agattagggt
    12781 agtatatgct atcctaattt atatctgggt agcatatact acccaaatat ctggatagca
    12841 tatgctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagca
    12901 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
    12961 tatgctatcc taatttatat ctgggtagca taggctatcc taatctatat ctgggtagca
    13021 tatgctatcc taatctatat ctgggtagta tatgctatcc taatctgtat ccgggtagca
    13081 tatgctatcc tcatgcatat acagtcagca tatgataccc agtagtagag tgggagtgct
    13141 atcctttgca tatgccgcca cctcccaagg gggcgtgaat tttcgctgct tgtccttttc
    13201 ctgctggttg ctcccattct taggtgaatt taaggaggcc aggctaaagc cgtcgcatgt
    13261 ctgattgctc accaggtaaa tgtcgctaat gttttccaac gcgagaaggt gttgagcgcg
    13321 gagctgagtg acgtgacaac atgggtatgc ccaattgccc catgttggga ggacgaaaat
    13381 ggtgacaaga cagatggcca gaaatacacc aacagcacgc atgatgtcta ctggggattt
    13441 attctttagt gcgggggaat acacggcttt taatacgatt gagggcgtct cctaacaagt
    13501 tacatcactc ctgcccttcc tcaccctcat ctccatcacc tccttcatct ccgtcatctc
    13561 cgtcatcacc ctccgcggca gccccttcca ccataggtgg aaaccaggga ggcaaatcta
    13621 ctccatcgtc aaagctgcac acagtcaccc tgatattgca ggtaggagcg ggctttgtca
    13681 taacaaggtc cttaatcgca tccttcaaaa cctcagcaaa tatatgagtt tgtaaaaaga
    13741 ccatgaaata acagacaatg gactccctta gcgggccagg ttgtgggccg ggtccagggg
    13801 ccattccaaa ggggagacga ctcaatggtg taagacgaca ttgtggaata gcaagggcag
    13861 ttcctcgcct taggttgtaa agggaggtct tactacctcc atatacgaac acaccggcga
    13921 cccaagttcc ttcgtcggta gtcctttcta cgtgactcct agccaggaga gctcttaaac
    13981 cttctgcaat gttctcaaat ttcgggttgg aacctccttg accacgatgc tttccaaacc
    14041 accctccttt tttgcgcctg cctccatcac cctgaccccg gggtccagtg cttgggcctt
    14101 ctcctgggtc atctgcgggg ccctgctcta tcgctcccgg gggcacgtca ggctcaccat
    14161 ctgggccacc ttcttggtgg tattcaaaat aatcggcttc ccctacaggg tggaaaaatg
    14221 gccttctacc tggagggggc ctgcgcggtg gagacccgga tgatgatgac tgactactgg
    14281 gactcctggg cctcttttct ccacgtccac gacctctccc cctggctctt tcacgacttc
    14341 ccaccctggc tctttcacgt cctctacccc ggcggcctcc actacctcct cgaccccggc
    14401 ctccactacc tcctcgaccc cggcctccac tgcctcctcg accccggcct ccacctcctg
    14461 ctcctgcccc tcctgctcct gcccctcctc ctgctcctgc ccctcctgcc cctcctgctc
    14521 ctgctcctgc tcctgttcca ccgtgggtcc ctttgcagcc aatgcr
//