LOCUS       sequence-464734-       13278 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses ABE8e(V106W) 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 13278)
  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 13278)
  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-CR033
FEATURES             Location/Qualifiers
     source          1..13278
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(830..850)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        835..1164
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1015..1150
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          2298..2363
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2493..2513
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2938..3072
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2975..2994)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3029..3048
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             3240..3873
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    3920..4045
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4542..4775
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4960..5004
                     /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    5289..5406
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5467..5678
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             5736..5756
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(6351..6374,6375..6377,6378..10451)
                     /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             6351..10451
                     /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             10497..10517
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             10527..10583
                     /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             10584..10982
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             11079..11312
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(11337..11355)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(11338..11355)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(11645..11664)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(11878..11897)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     11997..12019
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(12057..12075)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        12143..12247
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(12248..12316,12317..13108)
                     /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(12466..12485)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgtg gaatgtgtgt
      841 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
      901 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
      961 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     1021 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     1081 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     1141 tttggaggcc taggcttttg caaaaagctt ggacacaaga caggcttgcg agatatgttt
     1201 gagaatacca ctttatcccg cgtcagggag aggcagtgcg taaaaagacg cggactcatg
     1261 tgaaatactg gtttttagtg cgccagatct ctataatctc gcgcaaccta ttttcccctc
     1321 gaacactttt taagccgtag ataaacaggc tgggacactt cacatgagcg aaaaatacat
     1381 cgtcacctgg gacatgttgc agatccatgc acgtaaactc gcaagccgac tgatgccttc
     1441 tgaacaatgg aaaggcatta ttgccgtaag ccgtggcggt ctgtaccggg tgcgttactg
     1501 gcgcgtgaac tgggtattcg tcatgtcgat accgtttgta tttccagcta cgatcacgac
     1561 aaccagcgcg agcttaaagt gctgaaacgc gcagaaggcg atggcgaagg cttcatcgtt
     1621 attgatgacc tggtggatac cggtggtact gcggttgcga ttcgtgaaat gtatccaaaa
     1681 gcgcactttg tcaccatctt cgcaaaaccg gctggtcgtc cgctggttga tgactatgtt
     1741 gttgatatcc cgcaagatac ctggattgaa cagccgtggg atatgggcgt cgtattcgtc
     1801 ccgccaatct ccggtcgcta atcttttcaa cgcctggcac tgccgggcgt tgttcttttt
     1861 aacttcaggc gggttacaat agtttccagt aagtattctg gaggctgcat ccatgacaca
     1921 ggcaaacctg agcgaaaccc tgttcaaacc ccgctttaaa catcctgaaa cctcgacgct
     1981 agtccgccgc tttaatcacg gcgcacaacc gcctgtgcag tcggcccttg atggtaaaac
     2041 catccctcac tggtatcgca tgattaaccg tctgatgtgg atctggcgcg gcattgaccc
     2101 acgcgaaatc ctcgacgtcc aggcacgtat tgtgatgagc gatgccgaac gtaccgacga
     2161 tgatttatac gatacggtga ttggctaccg tggcggcaac tggatttatg agtgggcccc
     2221 ggatctttgt gaaggaacct tacttctgtg gtgtgacata attggacaaa ctacctacag
     2281 agatttaaag ctctaaggta aatataaaat ttttaagtgt ataatgtgtt aaactactga
     2341 ttctaattgt ttgtgtattt tagattccaa cctatggaac tgatgaatgg gagcagtggt
     2401 ggaatgcctt taatgaggaa aacctgtttt gctcagaaga aatgccatct agtgatgatg
     2461 aggctactgc tgactctcaa cattctactc ctccaaaaaa gaagagaaag gtagaagacc
     2521 ccaaggactt tccttcagaa ttgctaagtt ttttgagtca tgctgtgttt agtaatagaa
     2581 ctcttgcttg ctttgctatt tacaccacaa aggaaaaagc tgcactgcta tacaagaaaa
     2641 ttatggaaaa atattctgta acctttataa gtaggcataa cagttataat cataacatac
     2701 tgttttttct tactccacac aggcatagag tgtctgctat taataactat gctcaaaaat
     2761 tgtgtacctt tagcttttta atttgtaaag gggttaataa ggaatatttg atgtatagtg
     2821 ccttgactag agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     2881 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     2941 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3001 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     3061 catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact tcccacccca
     3121 taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg tgattcctct
     3181 gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa cttagtagtt
     3241 ggaagggcta attcactccc aaagaagaca agatatcctt gatctgtgga tctaccacac
     3301 acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact
     3361 gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa
     3421 taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga
     3481 gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga
     3541 gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc
     3601 tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc
     3661 ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag
     3721 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     3781 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     3841 gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc
     3901 gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc
     3961 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa
     4021 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga
     4081 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc
     4141 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg
     4201 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc
     4261 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac
     4321 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca
     4381 agcggccggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
     4441 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     4501 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     4561 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     4621 agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc tattgaggcg
     4681 caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc aagaatcctg
     4741 gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
     4801 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
     4861 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
     4921 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
     4981 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
     5041 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
     5101 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
     5161 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
     5221 agagacagat ccattcgatt agtgaacgga tctcgacggt atcgccaaat ggcagtattc
     5281 atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
     5341 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
     5401 aattttcggg tttattacag ggacagcaga gatccagttt ggatcgataa gcttgatatc
     5461 gaattcgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca
     5521 attgatccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact
     5581 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga
     5641 acgttctttt tcgcaacggg tttgccgcca gaacacaggg gatcccctgc agggccacca
     5701 tgaaacggac agccgacgga agcgagttcg agtcaccaaa gaagaagcgg aaagtctctg
     5761 aggtggagtt ttcccacgag tactggatga gacatgccct gaccctggcc aagagggcac
     5821 gggatgagag ggaggtgcct gtgggagccg tgctggtgct gaacaataga gtgatcggcg
     5881 agggctggaa cagagccatc ggcctgcacg acccaacagc ccatgccgaa attatggccc
     5941 tgagacaggg cggcctggtc atgcagaact acagactgat tgacgccacc ctgtacgtga
     6001 cattcgagcc ttgcgtgatg tgcgccggcg ccatgatcca ctctaggatc ggccgcgtgg
     6061 tgtttggatg gagaaattct aaaagaggcg ccgcaggctc cctgatgaac gtgctgaact
     6121 accccggcat gaatcaccgc gtcgaaatta ccgagggaat cctggcagat gaatgtgccg
     6181 ccctgctgtg cgatttctat cggatgccta gacaggtgtt caatgctcag aagaaggccc
     6241 agagctccat caactccgga ggatctagcg gaggctcctc tggctctgag acacctggca
     6301 caagcgagag cgcaacacct gaaagcagcg ggggcagcag cggggggtca gacaagaagt
     6361 acagcatcgg cctggccatc ggcaccaact ctgtgggctg ggccgtgatc accgacgagt
     6421 acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac agcatcaaga
     6481 agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc acccggctga
     6541 agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat ctgcaagaga
     6601 tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg gaagagtcct
     6661 tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac atcgtggacg
     6721 aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa ctggtggaca
     6781 gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg atcaagttcc
     6841 ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg gacaagctgt
     6901 tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc aacgccagcg
     6961 gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc
     7021 tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg attgccctga
     7081 gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat gccaaactgc
     7141 agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag atcggcgacc
     7201 agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg ctgagcgaca
     7261 tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg atcaagagat
     7321 acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg
     7381 agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc tacattgacg
     7441 gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa aagatggacg
     7501 gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag cagcggacct
     7561 tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc attctgcggc
     7621 ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag aagatcctga
     7681 ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga ttcgcctgga
     7741 tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg gtggacaagg
     7801 gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac ctgcccaacg
     7861 agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat aacgagctga
     7921 ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc ggcgagcaga
     7981 aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga
     8041 aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc ggcgtggaag
     8101 atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc aaggacaagg
     8161 acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg accctgacac
     8221 tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac ctgttcgacg
     8281 acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg ctgagccgga
     8341 agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat ttcctgaagt
     8401 ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc ctgaccttta
     8461 aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac gagcacattg
     8521 ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg aaggtggtgg
     8581 acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc gaaatggcca
     8641 gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg aagcggatcg
     8701 aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg gaaaacaccc
     8761 agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat atgtacgtgg
     8821 accaggaact ggacatcaac cggctgtccg actacgatgt ggaccatatc gtgcctcaga
     8881 gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac aagaaccggg
     8941 gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac tactggcggc
     9001 agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc aaggccgaga
     9061 gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg gtggaaaccc
     9121 ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact aagtacgacg
     9181 agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag ctggtgtccg
     9241 atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac caccacgccc
     9301 acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac cctaagctgg
     9361 aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg atcgccaaga
     9421 gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac atcatgaact
     9481 ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct ctgatcgaga
     9541 caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc accgtgcgga
     9601 aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag acaggcggct
     9661 tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc agaaagaagg
     9721 actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat tctgtgctgg
     9781 tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa gagctgctgg
     9841 ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt ctggaagcca
     9901 agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac tccctgttcg
     9961 agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag aagggaaacg
    10021 aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac tatgagaagc
    10081 tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag cacaagcact
    10141 acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc ctggccgacg
    10201 ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc atcagagagc
    10261 aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct gccgccttca
    10321 agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag gtgctggacg
    10381 ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac ctgtctcagc
    10441 tgggaggtga ctctggcggc tcaaaaagaa ccgccgacgg cagcgagttc gaaagcccca
    10501 agaagaagag gaaagtcggc tccggcgcaa caaacttctc tctgctgaaa caagccggag
    10561 atgtcgaaga gaatcctgga ccgatggcca agcctttgtc tcaagaagaa tccaccctca
    10621 ttgaaagagc aacggctaca atcaacagca tccccatctc tgaagactac agcgtcgcca
    10681 gcgcagctct ctctagcgac ggccgcatct tcactggtgt caatgtatat cattttactg
    10741 ggggaccttg tgcagaactc gtggtgctgg gcactgctgc tgctgcggca gctggcaacc
    10801 tgacttgtat cgtcgcgatc ggaaatgaga acaggggcat cttgagcccc tgcggacggt
    10861 gccgacaggt gcttctcgat ctgcatcctg ggatcaaagc catagtgaag gacagtgatg
    10921 gacagccgac ggcagttggg attcgtgaat tgctgccctc tggttatgtg tgggagggct
    10981 gaacgcgtcg atgagaggag tattcgtggt acctttaaga ccaatgactt acaaggcagc
    11041 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
    11101 acgaagacaa gatctgcttt ttgcttgtac tgggtctctc tggttagacc agatctgagc
    11161 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
    11221 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
    11281 acccttttag tcagtgtgga aaatctctag cagcatctag aattaattcc gtgtattcta
    11341 tagtgtcacc taaatcgtat gtgtatgata cataaggtta tgtattaatt gtagccgcgt
    11401 tctaacgaca atatgtacaa gcctaattgt gtagcatctg gcttactgaa gcagacccta
    11461 tcatctctct cgtaaactgc cgtcagagtc ggtttggttg gacgaacctt ctgagtttct
    11521 ggtaacgccg tcccgcaccc ggaaatggtc agcgaaccaa tcagcagggt catcgctagc
    11581 cagatcctct acgccggacg catcgtggcc ggcatcaccg gcgccacagg tgcggttgct
    11641 ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt cgggctcatg
    11701 agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact gttgggcgcc
    11761 atctccttgc atgcaccatt ccttgcggcg gcggtgctca acggcctcaa cctactactg
    11821 ggctgcttcc taatgcagga gtcgcataag ggagagcgtc gaatggtgca ctctcagtac
    11881 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
    11941 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
    12001 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
    12061 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
    12121 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
    12181 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
    12241 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
    12301 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
    12361 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
    12421 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
    12481 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
    12541 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
    12601 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
    12661 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
    12721 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
    12781 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
    12841 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
    12901 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
    12961 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
    13021 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
    13081 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
    13141 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
    13201 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
    13261 aaagatcaaa ggatcttc
//
