LOCUS       sequence-235834-       10036 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CAG promoter expression plasmid for 
            hSpCas9n(D10A)-UGI-NLS(SV40)-P2A-EGFP (BE3 control without 
            rAPOBEC1)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10036)
  AUTHORS   Grunewald J, Zhou R, Garcia SP, Iyer S, Lareau CA, Aryee MJ, Joung 
            JK
  TITLE     Transcriptome-wide off-target RNA editing induced by CRISPR-guided 
            DNA base editors.
  JOURNAL   Nature. 2019 Apr 17. pii: 10.1038/s41586-019-1161-z. doi: 
            10.1038/s41586-019-1161-z.
  PUBMED    30995674
REFERENCE   2  (bases 1 to 10036)
  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: "Nature. 
            2019 Apr 17. pii: 10.1038/s41586-019-1161-z. doi: 
            10.1038/s41586-019-1161-z."
COMMENT     Sequence Label: nCas9-UGI-NLS-P2A-EGFP (pJUL1001)
FEATURES             Location/Qualifiers
     source          1..10036
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(87..106)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    152..207
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(206..225)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(568..584)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(568..584)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(581..603)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    592..608
                     /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)."
     promoter        complement(join(616..622,623..640,641..646))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    661..682
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        741..936
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      787..922
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    942..1076
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(979..998)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1033..1052
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1144..1161)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1315..1903)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1395..1414)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2074..2865,2866..2934))
                     /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     2697..2716
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2935..3039)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3070..3449
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3451..3728
                     /label=chicken beta-actin promoter
     intron          3729..4745
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     4669..4691
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     4753..4772
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      4838..4847
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4844..8245,8246..8248,8249..8845,8846,8847..8849,
                     8850..8851,8852..8854,8855..8947)
                     /codon_start=1
                     /product="Streptococcus pyogenes Cas9 endonuclease with the
                     D1135V/R1335Q/T1337R mutations"
                     /label=Cas9 VQR
                     /note="has an altered recognition specificity for the 
                     protospacer adjacent motif (PAM) (Kleinstiver et al., 
                     2015)"
                     /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             join(4844..4870,4871..4873,4874..8947)
                     /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             8960..9208
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             9221..9241
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             9260..9316
                     /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             join(9317..9319,9320..9322,9323..10036)
                     /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             join(9317..9319,9320..9322,9323..10033)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(9362..9383)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(9623..9642)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     9970..9991
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 agcggccgca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat
       61 gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc
      121 atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata
      181 gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta
      241 aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc
      301 atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat
      361 tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg
      421 ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc
      481 ctcctctcct gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac
      541 ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
      601 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
      661 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
      721 cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact
      781 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
      841 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
      901 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc taacttgttt attgcagctt
      961 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     1021 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct
     1081 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     1141 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     1201 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     1261 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     1321 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     1381 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1441 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1501 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     1561 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     1621 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     1681 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     1741 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     1801 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     1861 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     1921 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     1981 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     2041 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     2101 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     2161 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     2221 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     2281 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     2341 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     2401 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     2461 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     2521 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     2581 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     2641 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     2701 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     2761 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     2821 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     2881 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     2941 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     3001 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     3061 acctgggtcg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
     3121 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     3181 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     3241 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     3301 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     3361 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     3421 tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc
     3481 tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt
     3541 gtgcagcgat gggggcgggg gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg
     3601 aggggcgggg cggggcgagg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc
     3661 gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc
     3721 ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc
     3781 gccgcccgcc ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc
     3841 cttctcctcc gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc
     3901 tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggg
     3961 gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct
     4021 gtgagcgctg cgggcgcggc gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg
     4081 cggccggggg cggtgccccg cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc
     4141 ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc
     4201 ccctgcaccc ccctccccga gttgctgagc acggcccggc ttcgggtgcg gggctccgta
     4261 cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg
     4321 gcggggcggg gccgcctcgg gccggggagg gctcggggga ggggcgcggc ggcccccgga
     4381 gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg taatcgtgcg
     4441 agagggcgca gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc
     4501 gccgcacccc ctctagcggg cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg
     4561 gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct tctccctctc cagcctcggg
     4621 gctgtccgcg gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct
     4681 ggcgtgtgac cggcggctct agagcctctg ctaaccatgt tcatgccttc ttctttttcc
     4741 tacagctcct gggcaacgtg ctggttattg tgctgtctca tcattttggc aaagaattct
     4801 gcagtcgacg gtaccgcggg cccgggatcc accggtcgcc accatggata aaaagtattc
     4861 tattggttta gccatcggca ctaattccgt tggatgggct gtcataaccg atgaatacaa
     4921 agtaccttca aagaaattta aggtgttggg gaacacagac cgtcattcga ttaaaaagaa
     4981 tcttatcggt gccctcctat tcgatagtgg cgaaacggca gaggcgactc gcctgaaacg
     5041 aaccgctcgg agaaggtata cacgtcgcaa gaaccgaata tgttacttac aagaaatttt
     5101 tagcaatgag atggccaaag ttgacgattc tttctttcac cgtttggaag agtccttcct
     5161 tgtcgaagag gacaagaaac atgaacggca ccccatcttt ggaaacatag tagatgaggt
     5221 ggcatatcat gaaaagtacc caacgattta tcacctcaga aaaaagctag ttgactcaac
     5281 tgataaagcg gacctgaggt taatctactt ggctcttgcc catatgataa agttccgtgg
     5341 gcactttctc attgagggtg atctaaatcc ggacaactcg gatgtcgaca aactgttcat
     5401 ccagttagta caaacctata atcagttgtt tgaagagaac cctataaatg caagtggcgt
     5461 ggatgcgaag gctattctta gcgcccgcct ctctaaatcc cgacggctag aaaacctgat
     5521 cgcacaatta cccggagaga agaaaaatgg gttgttcggt aaccttatag cgctctcact
     5581 aggcctgaca ccaaatttta agtcgaactt cgacttagct gaagatgcca aattgcagct
     5641 tagtaaggac acgtacgatg acgatctcga caatctactg gcacaaattg gagatcagta
     5701 tgcggactta tttttggctg ccaaaaacct tagcgatgca atcctcctat ctgacatact
     5761 gagagttaat actgagatta ccaaggcgcc gttatccgct tcaatgatca aaaggtacga
     5821 tgaacatcac caagacttga cacttctcaa ggccctagtc cgtcagcaac tgcctgagaa
     5881 atataaggaa atattctttg atcagtcgaa aaacgggtac gcaggttata ttgacggcgg
     5941 agcgagtcaa gaggaattct acaagtttat caaacccata ttagagaaga tggatgggac
     6001 ggaagagttg cttgtaaaac tcaatcgcga agatctactg cgaaagcagc ggactttcga
     6061 caacggtagc attccacatc aaatccactt aggcgaattg catgctatac ttagaaggca
     6121 ggaggatttt tatccgttcc tcaaagacaa tcgtgaaaag attgagaaaa tcctaacctt
     6181 tcgcatacct tactatgtgg gacccctggc ccgagggaac tctcggttcg catggatgac
     6241 aagaaagtcc gaagaaacga ttactccatg gaattttgag gaagttgtcg ataaaggtgc
     6301 gtcagctcaa tcgttcatcg agaggatgac caactttgac aagaatttac cgaacgaaaa
     6361 agtattgcct aagcacagtt tactttacga gtatttcaca gtgtacaatg aactcacgaa
     6421 agttaagtat gtcactgagg gcatgcgtaa acccgccttt ctaagcggag aacagaagaa
     6481 agcaatagta gatctgttat tcaagaccaa ccgcaaagtg acagttaagc aattgaaaga
     6541 ggactacttt aagaaaattg aatgcttcga ttctgtcgag atctccgggg tagaagatcg
     6601 atttaatgcg tcacttggta cgtatcatga cctcctaaag ataattaaag ataaggactt
     6661 cctggataac gaagagaatg aagatatctt agaagatata gtgttgactc ttaccctctt
     6721 tgaagatcgg gaaatgattg aggaaagact aaaaacatac gctcacctgt tcgacgataa
     6781 ggttatgaaa cagttaaaga ggcgtcgcta tacgggctgg ggacgattgt cgcggaaact
     6841 tatcaacggg ataagagaca agcaaagtgg taaaactatt ctcgattttc taaagagcga
     6901 cggcttcgcc aataggaact ttatgcagct gatccatgat gactctttaa ccttcaaaga
     6961 ggatatacaa aaggcacagg tttccggaca aggggactca ttgcacgaac atattgcgaa
     7021 tcttgctggt tcgccagcca tcaaaaaggg catactccag acagtcaaag tagtggatga
     7081 gctagttaag gtcatgggac gtcacaaacc ggaaaacatt gtaatcgaga tggcacgcga
     7141 aaatcaaacg actcagaagg ggcaaaaaaa cagtcgagag cggatgaaga gaatagaaga
     7201 gggtattaaa gaactgggca gccagatctt aaaggagcat cctgtggaaa atacccaatt
     7261 gcagaacgag aaactttacc tctattacct acaaaatgga agggacatgt atgttgatca
     7321 ggaactggac ataaaccgtt tatctgatta cgacgtcgat cacattgtac cccaatcctt
     7381 tttgaaggac gattcaatcg acaataaagt gcttacacgc tcggataaga accgagggaa
     7441 aagtgacaat gttccaagcg aggaagtcgt aaagaaaatg aagaactatt ggcggcagct
     7501 cctaaatgcg aaactgataa cgcaaagaaa gttcgataac ttaactaaag ctgagagggg
     7561 tggcttgtct gaacttgaca aggccggatt tattaaacgt cagctcgtgg aaacccgcca
     7621 aatcacaaag catgttgcac agatactaga ttcccgaatg aatacgaaat acgacgagaa
     7681 cgataagctg attcgggaag tcaaagtaat cactttaaag tcaaaattgg tgtcggactt
     7741 cagaaaggat tttcaattct ataaagttag ggagataaat aactaccacc atgcgcacga
     7801 cgcttatctt aatgccgtcg tagggaccgc actcattaag aaatacccga agctagaaag
     7861 tgagtttgtg tatggtgatt acaaagttta tgacgtccgt aagatgatcg cgaaaagcga
     7921 acaggagata ggcaaggcta cagccaaata cttcttttat tctaacatta tgaatttctt
     7981 taagacggaa atcactctgg caaacggaga gatacgcaaa cgacctttaa ttgaaaccaa
     8041 tggggagaca ggtgaaatcg tatgggataa gggccgggac ttcgcgacgg tgagaaaagt
     8101 tttgtccatg ccccaagtca acatagtaaa gaaaactgag gtgcagaccg gagggttttc
     8161 aaaggaatcg attcttccaa aaaggaatag tgataagctc atcgctcgta aaaaggactg
     8221 ggacccgaaa aagtacggtg gcttcgatag ccctacagtt gcctattctg tcctagtagt
     8281 ggcaaaagtt gagaagggaa aatccaagaa actgaagtca gtcaaagaat tattggggat
     8341 aacgattatg gagcgctcgt cttttgaaaa gaaccccatc gacttccttg aggcgaaagg
     8401 ttacaaggaa gtaaaaaagg atctcataat taaactacca aagtatagtc tgtttgagtt
     8461 agaaaatggc cgaaaacgga tgttggctag cgccggagag cttcaaaagg ggaacgaact
     8521 cgcactaccg tctaaatacg tgaatttcct gtatttagcg tcccattacg agaagttgaa
     8581 aggttcacct gaagataacg aacagaagca actttttgtt gagcagcaca aacattatct
     8641 cgacgaaatc atagagcaaa tttcggaatt cagtaagaga gtcatcctag ctgatgccaa
     8701 tctggacaaa gtattaagcg catacaacaa gcacagggat aaacccatac gtgagcaggc
     8761 ggaaaatatt atccatttgt ttactcttac caacctcggc gctccagccg cattcaagta
     8821 ttttgacaca acgatagatc gcaaacgata cacttctacc aaggaggtgc tagacgcgac
     8881 actgattcac caatccatca cgggattata tgaaactcgg atagatttgt cacagcttgg
     8941 gggtgactct ggtggttcta ctaatctgtc agatattatt gaaaaggaga ccggtaagca
     9001 actggttatc caggaatcca tcctcatgct cccagaggag gtggaagaag tcattgggaa
     9061 caagccggaa agcgatatac tcgtgcacac cgcctacgac gagagcaccg acgagaatgt
     9121 catgcttctg actagcgacg cccctgaata caagccttgg gctctggtca tacaggatag
     9181 caacggtgag aacaagatta agatgctctc tggtggttct cccaagaaga agaggaaagt
     9241 cggaggcggt ggaagcggcg caacaaactt ctctctgctg aaacaagccg gagatgtcga
     9301 agagaatcct ggaccgatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     9361 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     9421 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     9481 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     9541 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     9601 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     9661 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     9721 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     9781 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     9841 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     9901 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     9961 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
    10021 cgagctgtac aagtga
//
