LOCUS       sequence-465064-       13937 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses evoCDA1 BE4max base editor in a human T cell optimized 
            lentiviral transfer plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13937)
  AUTHORS   Schmidt R, Ward CC, Dajani R, Armour-Garb Z, Ota M, Allain V, 
            Hernandez R, Layeghi M, Xing G, Goudy L, Dorovskyi D, Wang C, Chen 
            YY, Ye CJ, Shy BR, Gilbert LA, Eyquem J, Pritchard JK, Dodgson SE, 
            Marson A
  TITLE     Base-editing mutagenesis maps alleles to tune human T cell 
            functions.
  JOURNAL   Nature. 2024 Jan;625(7996):805-812. doi: 10.1038/s41586-023-06835-6.
            Epub 2023 Dec 13.
  PUBMED    38093011
REFERENCE   2  (bases 1 to 13937)
  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/s41586-023-06835-6"; journalName: "Nature"; date: 
            "2024-01"; volume: "625"; issue: "7996"; pages: "805-812"
COMMENT     Sequence Label: pLenti-CR029
FEATURES             Location/Qualifiers
     source          1..13937
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      2..590
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     491..510
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     744..761
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(831..851)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        836..1165
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1016..1151
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          2299..2364
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2494..2514
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2939..3073
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2976..2995)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3030..3049
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             3241..3874
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    3921..4046
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4543..4776
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4961..5005
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    5290..5407
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5468..5679
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      5695..5704
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5740..5760
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(6481..6504,6505..6507,6508..10581)
                     /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             6481..10581
                     /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             10612..10860
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             10891..11139
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             11182..11202
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11212..11268
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             11269..11667
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             11739..11972
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(11997..12015)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(11998..12015)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(12305..12324)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(12538..12557)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     12657..12679
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(12717..12735)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        12803..12907
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(12908..12976,12977..13768)
                     /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     complement(13126..13145)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cttgagatcc tttttttttg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
       61 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
      121 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
      181 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
      241 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
      301 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
      361 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
      421 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
      481 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
      541 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
      601 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
      661 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
      721 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
      781 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgt ggaatgtgtg
      841 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
      901 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
      961 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     1021 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     1081 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     1141 ttttggaggc ctaggctttt gcaaaaagct tggacacaag acaggcttgc gagatatgtt
     1201 tgagaatacc actttatccc gcgtcaggga gaggcagtgc gtaaaaagac gcggactcat
     1261 gtgaaatact ggtttttagt gcgccagatc tctataatct cgcgcaacct attttcccct
     1321 cgaacacttt ttaagccgta gataaacagg ctgggacact tcacatgagc gaaaaataca
     1381 tcgtcacctg ggacatgttg cagatccatg cacgtaaact cgcaagccga ctgatgcctt
     1441 ctgaacaatg gaaaggcatt attgccgtaa gccgtggcgg tctgtaccgg gtgcgttact
     1501 ggcgcgtgaa ctgggtattc gtcatgtcga taccgtttgt atttccagct acgatcacga
     1561 caaccagcgc gagcttaaag tgctgaaacg cgcagaaggc gatggcgaag gcttcatcgt
     1621 tattgatgac ctggtggata ccggtggtac tgcggttgcg attcgtgaaa tgtatccaaa
     1681 agcgcacttt gtcaccatct tcgcaaaacc ggctggtcgt ccgctggttg atgactatgt
     1741 tgttgatatc ccgcaagata cctggattga acagccgtgg gatatgggcg tcgtattcgt
     1801 cccgccaatc tccggtcgct aatcttttca acgcctggca ctgccgggcg ttgttctttt
     1861 taacttcagg cgggttacaa tagtttccag taagtattct ggaggctgca tccatgacac
     1921 aggcaaacct gagcgaaacc ctgttcaaac cccgctttaa acatcctgaa acctcgacgc
     1981 tagtccgccg ctttaatcac ggcgcacaac cgcctgtgca gtcggccctt gatggtaaaa
     2041 ccatccctca ctggtatcgc atgattaacc gtctgatgtg gatctggcgc ggcattgacc
     2101 cacgcgaaat cctcgacgtc caggcacgta ttgtgatgag cgatgccgaa cgtaccgacg
     2161 atgatttata cgatacggtg attggctacc gtggcggcaa ctggatttat gagtgggccc
     2221 cggatctttg tgaaggaacc ttacttctgt ggtgtgacat aattggacaa actacctaca
     2281 gagatttaaa gctctaaggt aaatataaaa tttttaagtg tataatgtgt taaactactg
     2341 attctaattg tttgtgtatt ttagattcca acctatggaa ctgatgaatg ggagcagtgg
     2401 tggaatgcct ttaatgagga aaacctgttt tgctcagaag aaatgccatc tagtgatgat
     2461 gaggctactg ctgactctca acattctact cctccaaaaa agaagagaaa ggtagaagac
     2521 cccaaggact ttccttcaga attgctaagt tttttgagtc atgctgtgtt tagtaataga
     2581 actcttgctt gctttgctat ttacaccaca aaggaaaaag ctgcactgct atacaagaaa
     2641 attatggaaa aatattctgt aacctttata agtaggcata acagttataa tcataacata
     2701 ctgttttttc ttactccaca caggcataga gtgtctgcta ttaataacta tgctcaaaaa
     2761 ttgtgtacct ttagcttttt aatttgtaaa ggggttaata aggaatattt gatgtatagt
     2821 gccttgacta gagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     2881 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     2941 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3001 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3061 tcatgtctgg atcaactgga taactcaagc taaccaaaat catcccaaac ttcccacccc
     3121 ataccctatt accactgcca attacctgtg gtttcattta ctctaaacct gtgattcctc
     3181 tgaattattt tcattttaaa gaaattgtat ttgttaaata tgtactacaa acttagtagt
     3241 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
     3301 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
     3361 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
     3421 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
     3481 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
     3541 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
     3601 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
     3661 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
     3721 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     3781 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     3841 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
     3901 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
     3961 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
     4021 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
     4081 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
     4141 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
     4201 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     4261 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     4321 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     4381 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     4441 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     4501 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     4561 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     4621 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     4681 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     4741 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     4801 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     4861 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     4921 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     4981 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     5041 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     5101 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     5161 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     5221 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccaaa tggcagtatt
     5281 catccacaat tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt
     5341 agacataata gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca
     5401 aaattttcgg gtttattaca gggacagcag agatccagtt tggatcgata agcttgatat
     5461 cgaattcggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc
     5521 aattgatccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac
     5581 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg
     5641 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg ggatcccctg cagggccacc
     5701 atgggcaaac ggacagccga cggaagcgag ttcgagtcac caaagaagaa gcggaaagtc
     5761 agtaccgacg ccgagtacgt gcggatccac gagaagctgg atatctatac attcaagaag
     5821 cagtttagca acaataagaa gtccgtgtct cacagatgct acgtgctgtt cgagctgaag
     5881 cggagaggag agaggcgcgc ctgtttttgg ggctatgccg tgaacaagcc acagtctgga
     5941 accgagaggg gaatccacgc agagatcttc agcatcagga aggtggagga gtacctgcgc
     6001 gacaaccccg gccagtttac aatcaattgg tatagctcct ggagcccttg cgccgattgt
     6061 gccgagaaga tcctggagtg gtacaaccag gagctgaggg gcaatggcca caccctgaag
     6121 atctgggtgt gcaagctgta ctatgagaag aacgccagga atcagatcgg cctgtggaac
     6181 ctgcgcgaca atggcgtggg cctgaacgtg atggtgtccg agcactatca gtgctgtcgc
     6241 aagatcttta tccagtctag ccacaatcag ctgaacgaga atcggtggct ggagaagaca
     6301 ctgaagagag ccgagaagcg gagaagcgag ctgtccatca tgtttcaggt gaagatcctg
     6361 cacaccacaa agtctcccgc cgtgtctggc ggatctagcg gaggatcctc tggcagcgag
     6421 acaccaggaa caagcgagtc agcaacacca gagagcagtg gcggcagcag cggcggcagc
     6481 gacaagaagt acagcatcgg cctggccatc ggcaccaact ctgtgggctg ggccgtgatc
     6541 accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac
     6601 agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc
     6661 acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat
     6721 ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg
     6781 gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac
     6841 atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa
     6901 ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg
     6961 atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg
     7021 gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc
     7081 aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg
     7141 ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg
     7201 attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat
     7261 gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag
     7321 atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg
     7381 ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg
     7441 atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag
     7501 cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc
     7561 tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa
     7621 aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag
     7681 cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc
     7741 attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag
     7801 aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga
     7861 ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg
     7921 gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac
     7981 ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat
     8041 aacgagctga ccaaagtgaa atacgtgact gagggaatga gaaagcccgc cttcctgagc
     8101 ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg
     8161 aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc
     8221 ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc
     8281 aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg
     8341 accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac
     8401 ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg
     8461 ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat
     8521 ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc
     8581 ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac
     8641 gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg
     8701 aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc
     8761 gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg
     8821 aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg
     8881 gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat
     8941 atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt ggaccatatc
     9001 gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac
     9061 aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac
     9121 tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc
     9181 aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg
     9241 gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact
     9301 aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag
     9361 ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac
     9421 caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac
     9481 cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg
     9541 atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac
     9601 atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct
     9661 ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc
     9721 accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag
     9781 acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc
     9841 agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat
     9901 tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa
     9961 gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt
    10021 ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac
    10081 tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag
    10141 aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac
    10201 tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag
    10261 cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc
    10321 ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc
    10381 atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct
    10441 gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag
    10501 gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac
    10561 ctgtctcagc tgggaggtga cagcggcggg agcggcggga gcggggggag cactaatctg
    10621 agcgacatca ttgagaagga gactgggaaa cagctggtca ttcaggagtc catcctgatg
    10681 ctgcctgagg aggtggagga agtgatcggc aacaagccag agtctgacat cctggtgcac
    10741 accgcctacg acgagtccac agatgagaat gtgatgctgc tgacctctga cgcccccgag
    10801 tataagcctt gggccctggt catccaggat tctaacggcg agaataagat caagatgctg
    10861 agcggaggat ccggaggatc tggaggcagc accaacctgt ctgacatcat cgagaaggag
    10921 acaggcaagc agctggtcat ccaggagagc atcctgatgc tgcccgaaga agtcgaagaa
    10981 gtgatcggaa acaagcctga gagcgatatc ctggtccata ccgcctacga cgagagtacc
    11041 gacgaaaatg tgatgctgct gacatccgac gccccagagt ataagccctg ggctctggtc
    11101 atccaggatt ccaacggaga gaacaaaatc aaaatgctgt ctggcggctc aaaaagaacc
    11161 gccgacggca gcgaattcga gcccaagaag aagaggaaag tcggatccgg cgcaacaaac
    11221 ttctctctgc tgaaacaagc cggagatgtc gaagagaatc ctggaccgat ggccaagcct
    11281 ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa cagcatcccc
    11341 atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg catcttcact
    11401 ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt gctgggcact
    11461 gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa tgagaacagg
    11521 ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca tcctgggatc
    11581 aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg tgaattgctg
    11641 ccctctggtt atgtgtggga gggctaaggt acctttaaga ccaatgactt acaaggcagc
    11701 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
    11761 acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc
    11821 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
    11881 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
    11941 acccttttag tcagtgtgga aaatctctag cagcatctag aattaattcc gtgtattcta
    12001 tagtgtcacc taaatcgtat gtgtatgata cataaggtta tgtattaatt gtagccgcgt
    12061 tctaacgaca atatgtacaa gcctaattgt gtagcatctg gcttactgaa gcagacccta
    12121 tcatctctct cgtaaactgc cgtcagagtc ggtttggttg gacgaacctt ctgagtttct
    12181 ggtaacgccg tcccgcaccc ggaaatggtc agcgaaccaa tcagcagggt catcgctagc
    12241 cagatcctct acgccggacg catcgtggcc ggcatcaccg gcgccacagg tgcggttgct
    12301 ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt cgggctcatg
    12361 agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact gttgggcgcc
    12421 atctccttgc atgcaccatt ccttgcggcg gcggtgctca acggcctcaa cctactactg
    12481 ggctgcttcc taatgcagga gtcgcataag ggagagcgtc gaatggtgca ctctcagtac
    12541 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
    12601 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
    12661 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
    12721 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
    12781 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
    12841 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
    12901 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
    12961 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
    13021 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
    13081 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
    13141 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
    13201 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
    13261 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
    13321 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
    13381 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
    13441 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
    13501 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
    13561 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
    13621 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
    13681 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
    13741 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
    13801 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
    13861 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
    13921 aaagatcaaa ggatctt
//
