LOCUS       sequence-251688-       10439 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CAG promoter expression plasmid for 
            NLS-hSpCas9n(D10A)-NLS-SH3-3xFLAG-NLS-UGI-P2A-EGFP (Target-AID 
            without pmCDA1). .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10439)
  AUTHORS   Grunewald J, Zhou R, Iyer S, Lareau CA, Garcia SP, Aryee MJ, Joung 
            JK
  TITLE     CRISPR DNA base editors with reduced RNA off-target and self-editing
            activities.
  JOURNAL   Nat Biotechnol. 2019 Sep 2. pii: 10.1038/s41587-019-0236-6. doi: 
            10.1038/s41587-019-0236-6.
  PUBMED    31477922
REFERENCE   2  (bases 1 to 10439)
  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"; journalName: "Nat 
            Biotechnol. 2019 Sep 2. pii: 10.1038/s41587-019-0236-6. doi: 
            10.1038/s41587-019-0236-6."
COMMENT     Sequence Label: nCas9-UGI control for Target-AID (pRZ1047)
FEATURES             Location/Qualifiers
     source          1..10439
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     220..239
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     473..490
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    558..692
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(582..601)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     636..655
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(698..893)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      712..847
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    952..973
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(988..993,994..1011,1012..1018)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1026..1042
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     1031..1053
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1050..1066
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1050..1066
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1409..1428
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    1427..1482
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     1528..1547
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(join(1634..2347,2348..2350,2351..2353))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(1637..2347,2348..2350,2351..2353))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1679..1700)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2028..2047
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2287..2308
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(2354..2410)
                     /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             complement(2429..2680)
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="MTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHT
                     AYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             complement(2687..2707)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(2717..2782)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(2990..3010)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(join(3023..7096,7097..7099,7100..7126))
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(3023..7126)
                     /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             complement(7151..7171)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      7174..7183
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(7249..7268)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          7276..8292
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(7330..7352)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(8293..8570)
                     /label=chicken beta-actin promoter
     enhancer        8572..8951
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8982..9086
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9087..9155,9156..9947)
                     /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(9305..9324)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
       61 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
      121 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
      181 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
      241 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
      301 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
      361 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
      421 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
      481 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
      541 gattcattaa tgcagcggat ccagacatga taagatacat tgatgagttt ggacaaacca
      601 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
      661 ttgtaaccat tataagctgc aataaacaag ttagcttttt gcaaaagcct aggcctccaa
      721 aaaagcctcc tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat
      781 aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg
      841 gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga tgcggatccg
      901 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
      961 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     1021 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     1081 gcttgggctg caggtcgagg gatctccata agagaagagg gacagctatg actgggagta
     1141 gtcaggagag gaggaaaaat ctggctagta aaacatgtaa ggaaaatttt agggatgtta
     1201 aagaaaaaaa taacacaaaa caaaatataa aaaaaatcta acctcaagtc aaggcttttc
     1261 tatggaataa ggaatggaca gcagggggct gtttcatata ctgatgacct ctttatagcc
     1321 aacctttgtt catggcagcc agcatatggg catatgttgc caaactctaa accaaatact
     1381 cattctgatg ttttaaatga tttgccctcc catatgtcct tccgagtgag agacacaaaa
     1441 aattccaaca cactattgca atgaaaataa atttccttta ttagccagaa gtcagatgct
     1501 caaggggctt catgatgtcc ccataatttt tggcagaggg aaaaagatct cagtggtatt
     1561 tgtgagccag ggcattggcc acaccagcca ccaccttctg ataggcagcc tgcacctgag
     1621 gagtgcggcc gcttcacttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca
     1681 cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc agggcggact
     1741 gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct
     1801 gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt
     1861 tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt
     1921 tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct
     1981 cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc
     2041 cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga
     2101 agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg ccgtaggtca
     2161 gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgaacttca
     2221 gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc
     2281 cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc
     2341 ccttgctcac catcggtcca ggattctctt cgacatctcc ggcttgtttc agcagagaga
     2401 agtttgttgc gccgcttcca ccgcctccta gcatcttgat cttgttctct ccattagagt
     2461 cctgaatcac cagagcccag ggtttgtact ctggggcatc tgacgtgagc agcatcacgt
     2521 tctcatcggt gctctcatca tacgcagtgt gtacgagaat gtcactttcc ggcttattcc
     2581 caatgacctc ctcaacttct tcaggaagca tgagtatcga ctcttggatg accaactgtt
     2641 tgcctgtttc cttctctatg atgtcggaaa ggttggtcat ggtaccgact tttcttttct
     2701 tcttgggacc ccctgattta tcgtcgtcgt ctttgtagtc gatgtcgtgg tctttgtaat
     2761 ccccatcatg gtctttgtag tcaccggaat atttctccac atatggcacc agaatcatgc
     2821 cccttttccc ttcgctatcc tcggcattcc accattgttc ttcgggctta tctctgatgc
     2881 ggagaatatc gcccttctta aaggggagat cctcctcgtc attcccattg aagtcaaaca
     2941 gagctcgtac gtattctgct gagcctccac cgccggtacc tcctccaccc accttcctct
     3001 tcttcttggg gtcagccctg ctgtctccac cgagctgaga gaggtcgatt cttgtttcat
     3061 agagccccgt aattgactga tgaatcagtg tggcgtccag gacctccttt gtagaggtgt
     3121 accgctttct gtctatggtg gtgtcgaagt acttgaaggc tgcaggcgcg cccaagttgg
     3181 tcagagtaaa caagtggata atgttttctg cctgctccct gatgggctta tccctgtgct
     3241 tattgtaagc agaaagcacc ttatcgaggt tagcgtcggc gaggatcact cttttggaga
     3301 attcgcttat ttgctcgatg atctcatcaa ggtagtgttt gtgttgttcc acgaacagct
     3361 gcttctgctc attatcttcg ggagaccctt tgagcttttc atagtggctg gccagataca
     3421 agaaattaac gtatttagag ggcagtgcca gctcgttacc tttctgcagc tcgcccgcac
     3481 tagcgagcat tcgtttccgg ccgttttcaa gctcaaagag agagtacttg ggaagcttaa
     3541 tgatgaggtc ttttttgacc tctttatatc ctttcgcctc gagaaagtcg atggggtttt
     3601 tttcgaagct tgatcgctcc atgattgtga tgcccagcag ttccttgacg cttttgagtt
     3661 ttttagactt ccctttctcc actttggcca caaccagtac actgtaagcg actgtaggag
     3721 aatcgaatcc gccgtatttc ttggggtccc aatctttttt gcgtgcgatc agcttgtcgc
     3781 tgttcctttt cgggaggata ctttccttgg agaagcctcc ggtctgtact tcggtctttt
     3841 taacgatgtt cacctgcggc atggacagga ccttccggac tgtcgcgaaa tccctaccct
     3901 tgtcccacac gatttctcct gtttctccgt ttgtttcgat aagtggtcgc ttccgaatct
     3961 ctccattggc cagtgtaatc tcggtcttga aaaaattcat aatattgctg taaaagaagt
     4021 acttagcggt ggccttgcct atttcctgct cagactttgc gatcattttc ctaacatcgt
     4081 acactttata gtctccgtaa acaaattcag attcaagctt gggatatttt ttgataagtg
     4141 cagtgcctac cactgcattc aggtaggcat catgcgcatg gtggtaattg ttgatctctc
     4201 tcaccttata aaactgaaag tcctttctga aatctgagac cagcttagac ttcagagtaa
     4261 taactttcac ctctcgaatc agtttgtcat tttcatcgta cttggtgttc atgcgtgaat
     4321 cgagaatttg ggccacgtgc ttggtgatct ggcgtgtctc aacaagctgc cttttgatga
     4381 agccggcttt atccaactca gacaggccac ctcgttcagc cttagtcaga ttatcgaact
     4441 tccgttgtgt gatcagtttg gcgttcagca gctgccgcca ataatttttc attttcttga
     4501 caacttcttc tgaggggacg ttatcactct tccctctatt tttatcggat cttgtcaaca
     4561 ctttattatc aatagaatca tctttgagaa aagactgggg cacgatatga tccacgtcgt
     4621 agtcggagag ccgattgatg tccagttcct gatccacgta catgtccctg ccgttctgca
     4681 ggtagtacag gtagagcttc tcattctgaa gctgggtgtt ttcaactggg tgttccttaa
     4741 ggatttggga ccccagttct tttataccct cttcaatcct cttcatcctt tccctactgt
     4801 tcttctgtcc cttctgggta gtttggttct ctcgggccat ctcgataacg atattctcgg
     4861 gcttatgcct tcccattact ttgacgagtt catccacgac cttaacggtc tgcagtattc
     4921 cctttttgat agctgggcta cctgcaagat tagcgatgtg ctcgtgaaga ctgtccccct
     4981 ggccagaaac ttgtgctttc tggatgtcct ccttaaaggt gagagagtca tcatggatca
     5041 actgcatgaa gttccggttg gcaaatccat cggacttaag aaaatccagg attgtctttc
     5101 cactctgctt gtctcggatc ccattgatca gttttcttga cagccgcccc catcctgtat
     5161 atcggcgcct cttgagctgt ttcatgactt tgtcgtcgaa gagatgagcg taagttttca
     5221 agcgttcttc aatcatctcc ctatcttcaa acaacgtaag ggtgaggaca atgtcctcaa
     5281 gaatgtcctc gttctcctca ttgtccagga agtccttgtc tttaatgatt ttcaggagat
     5341 cgtgatacgt tcccagggat gcgttgaagc gatcctccac tccgctgatt tcaacagagt
     5401 cgaaacattc aatctttttg aaatagtctt ctttgagctg tttcacggta actttccggt
     5461 tcgtcttgaa gaggaggtcc acgatagctt tcttctgctc tccagacagg aatgctggct
     5521 ttctcatccc ttctgtgacg tatttgacct tggtgagctc gttataaact gtgaagtact
     5581 cgtacagcag agagtgttta ggaagcacct tttcgttagg cagattttta tcaaagttag
     5641 tcatcctttc gatgaaggac tgggcagagg cccccttatc cacgacttcc tcgaagttcc
     5701 agggagtgat ggtctcttct gatttgcgag tcatccacgc gaatctggaa tttccccggg
     5761 cgagggggcc tacatagtag ggtatccgaa atgtgaggat tttctcaatc ttttccctgt
     5821 tatctttcaa aaaggggtag aaatcctctt gccgcctgag gatagcgtgc agttcgccca
     5881 ggtgaatctg gtgggggatg cttccattgt cgaaagtgcg ctgtttgcgc aacagatctt
     5941 ctctgttaag ctttaccagc agctcctcgg tgccgtccat tttttccaag atgggcttaa
     6001 taaatttgta aaattcctcc tggcttgctc cgccgtcaat gtatccggcg tagccatttt
     6061 tagactgatc gaagaaaatt tccttgtact tctcaggcag ttgctgtctg acaagggcct
     6121 tcagcaaagt caagtcttgg tggtgctcat catagcgctt gatcatacta gcgctcagcg
     6181 gagctttggt gatctccgtg ttcactcgca gaatatcact cagcagaatg gcgtctgaca
     6241 ggttctttgc cgccaaaaaa aggtctgcgt actggtcgcc gatctgggcc agcagattgt
     6301 cgagatcatc atcgtaggtg tctttgctca gttgaagctt ggcatcttcg gccaggtcga
     6361 agttagattt aaagttgggg gtcagcccga gtgacagggc gataagatta ccaaacaggc
     6421 cgttcttctt ctccccaggg agctgtgcga tgaggttttc gagccgccgg gatttggaca
     6481 gcctagcgct caggattgct ttggcgtcaa ctccggatgc gttgatcggg ttctcttcga
     6541 aaagctgatt gtaagtctga accagttgga taaagagttt gtcgacatcg ctgttgtctg
     6601 ggttcaggtc cccctcgatg aggaagtgtc cccgaaattt gatcatatgc gccagcgcga
     6661 gatagatcaa ccgcaagtca gccttatcag tactgtctac aagcttcttc ctcagatgat
     6721 atatggttgg gtacttttca tggtacgcca cctcgtccac gatattgcca aagattgggt
     6781 ggcgctcgtg ctttttatcc tcctccacca aaaaggactc ctccagccta tggaagaaag
     6841 agtcatccac cttagccatc tcattactaa agatctcctg caggtagcag atccgattct
     6901 ttctgcgggt atatctgcgc cgtgctgttc ttttgagccg cgtggcttcg gccgtctccc
     6961 cggagtcgaa caggagggcg ccaatgaggt tcttctttat gctgtggcga tcggtattgc
     7021 ccagaacttt gaattttttg ctcggcacct tgtactcgtc cgtaatgacg gcccaaccga
     7081 cgctgtttgt gccgatagcg agcccaatgg agtacttctt gtccattgcc gcaggaactc
     7141 cgtggattcc gactttacgc ttcttcttcg gtgccatggt ggcgaccggt ggatcccggg
     7201 cccgcggtac cgtcgactgc agaattcttt gccaaaatga tgagacagca caataaccag
     7261 cacgttgccc aggagctgta ggaaaaagaa gaaggcatga acatggttag cagaggctct
     7321 agagccgccg gtcacacgcc agaagccgaa ccccgccctg ccccgtcccc cccgaaggca
     7381 gccgtccccc cgcggacagc cccgaggctg gagagggaga aggggacggc ggcgcggcga
     7441 cgcacgaagg ccctccccgc ccatttcctt cctgccggcg ccgcaccgct tcgccccgcg
     7501 cccgctagag ggggtgcggc ggcgcctccc agatttcggc tccgcacaga tttgggacaa
     7561 aggaagtccc tgcgccctct cgcacgatta ccataaaagg caatggctgc ggctcgccgc
     7621 gcctcgacag ccgccggcgc tccgggggcc gccgcgcccc tcccccgagc cctccccggc
     7681 ccgaggcggc cccgccccgc ccggcacccc cacctgccgc caccccccgc ccggcacggc
     7741 gagccccgcg ccacgccccg tacggagccc cgcacccgaa gccgggccgt gctcagcaac
     7801 tcggggaggg gggtgcaggg ggggttgcag cccgaccgac gcgcccacac cccctgctca
     7861 cccccccacg cacacacccc gcacgcagcc tttgttcccc tcgcagcccc ccccgcaccg
     7921 cggggcaccg cccccggccg cgctcccctc gcgcacactg cggagcgcac aaagccccgc
     7981 gccgcgcccg cagcgctcac agccgccggg cagcgcggag ccgcacgcgg cgctccccac
     8041 gcacacacac acgcacgcac cccccgagcc gctccccccg cacaaagggc cctcccggag
     8101 cccctcaagg ctttcacgca gccacagaaa agaaacaagc cgtcattaaa ccaagcgcta
     8161 attacagccc ggaggagaag ggccgtcccg cccgctcacc tgtgggagta acgcggtcag
     8221 tcagagccgg ggcgggcggc gcgaggcggc ggcggagcgg ggcacggggc gaaggcagcg
     8281 cgcagcgact cccgcccgcc gcgcgcttcg ctttttatag ggccgccgcc gccgccgcct
     8341 cgccataaaa ggaaactttc ggagcgcgcc gctctgattg gctgccgccg cacctctccg
     8401 cctcgccccg ccccgcccct cgccccgccc cgccccgcct ggcgcgcgcc cccccccccc
     8461 ccccgccccc atcgctgcac aaaataatta aaaaataaat aaatacaaaa ttgggggtgg
     8521 ggaggggggg gagatgggga gagtgaagca gaacgtgggg ctcacctcga ccatggtaat
     8581 agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg
     8641 ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat
     8701 atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg
     8761 tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg
     8821 gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa
     8881 ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat aactagtcaa
     8941 taatcaatgt cgacccaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     9001 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     9061 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     9121 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     9181 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     9241 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     9301 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     9361 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     9421 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     9481 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     9541 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     9601 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     9661 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     9721 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     9781 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     9841 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     9901 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     9961 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
    10021 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
    10081 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
    10141 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
    10201 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
    10261 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
    10321 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
    10381 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacg
//
