LOCUS       sequence-254665-       10678 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CAG promoter expression plasmid for hA3A-XTEN 
            linker-hSpCas9n(D10A)-UGI-NLS-P2A-EGFP..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10678)
  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 10678)
  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: hA3A-BE3 (pRZ215)
FEATURES             Location/Qualifiers
     source          1..10678
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(441..1232,1233..1301))
                     /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     1064..1083
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1302..1406)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        1437..1816
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1818..2095
                     /label=chicken beta-actin promoter
     intron          2096..3112
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3036..3058
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3120..3139
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      3205..3214
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3211..3807
                     /codon_start=1
                     /gene="human APOBEC3A"
                     /gene_synonym="A3A"
                     /product="human cytidine deaminase (C to U editing enzyme) 
                     that efficiently edits methylated cytosines (Wang et al., 
                     2018)"
                     /label=APOBEC3A
                     /note="also known as hA3A"
                     /translation="MEASPASGPRHLMDPHIFTSNFNNGIGRHKTYLCYEVERLDNGTS
                     VKMDQHRGFLHNQAKNLLCGFYGRHAELRFLDLVPSLQLDPAQIYRVTWFISWSPCFSW
                     GCAGEVRAFLQENTHVRLRIFAARIYDYDPLYKEALQMLRDAGAQVSIMTYDEFKHCWD
                     TFVDHQGCPFQPWDGLDEHSQALSGRLRAILQNQGN"
     CDS             join(3856..7254,7255..7257,7258..7854,7855,7856..7858,
                     7859..7860,7861..7863,7864..7956)
                     /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="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             join(3856..3879,3880..3882,3883..7956)
                     /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             7969..8217
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             8230..8250
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             8269..8325
                     /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(8326..8328,8329..8331,8332..9045)
                     /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(8326..8328,8329..8331,8332..9042)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(8371..8392)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(8632..8651)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8979..9000
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(9132..9151)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    9197..9252
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(9251..9270)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(9613..9629)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9613..9629)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9626..9648)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9637..9653
                     /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(9661..9667,9668..9685,9686..9691))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9706..9727
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        9786..9981
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      9832..9967
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    9987..10121
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(10024..10043)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     10078..10097
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(10189..10206)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(10360..10678,1..270))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10440..10459)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
       61 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      121 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
      181 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
      241 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
      301 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
      361 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
      421 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
      481 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
      541 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
      601 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
      661 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
      721 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
      781 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
      841 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
      901 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
      961 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     1021 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     1081 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     1141 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     1201 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     1261 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     1321 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     1381 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgggtcgaca
     1441 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     1501 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     1561 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     1621 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     1681 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     1741 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     1801 catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc
     1861 cccctcccca cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg
     1921 ggcggggggg gggggggggc gcgcgccggg gggggggggg gggggsgggg ggsggggsgg
     1981 ggsgrggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt
     2041 tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt
     2101 cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg
     2161 gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg
     2221 ctgtaattag cgcttggttt aatgacggct tgtttctttt ctgtggctgc gtgaaagcct
     2281 tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg
     2341 tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg
     2401 gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg
     2461 tgccccgcgg tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt
     2521 gggggggtga gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc
     2581 tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc
     2641 ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc
     2701 gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg ccggcggctg
     2761 tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg
     2821 acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc
     2881 tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct
     2941 tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg
     3001 ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg
     3061 cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg
     3121 caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattctgca gtcgacggta
     3181 ccgcgggccc gggatccacc ggtcgccacc atggaagcca gcccagcatc cgggcccaga
     3241 cacttgatgg atccacacat attcacttcc aactttaaca atggcattgg aaggcataag
     3301 acctacctgt gctacgaagt ggagcgcctg gacaatggca cctcggtcaa gatggaccag
     3361 cacaggggct ttctacacaa ccaggctaag aatcttctct gtggctttta cggccgccat
     3421 gcggagctgc gcttcttgga cctggttcct tctttgcagt tggacccggc ccagatctac
     3481 agggtcactt ggttcatctc ctggagcccc tgcttctcct ggggctgtgc cggggaagtg
     3541 cgtgcgttcc ttcaggagaa cacacacgtg agactgcgta tcttcgctgc ccgcatctat
     3601 gattacgacc ccctatataa ggaggcactg caaatgctgc gggatgctgg ggcccaagtc
     3661 tccatcatga cctacgatga atttaagcac tgctgggaca cctttgtgga ccaccaggga
     3721 tgtcccttcc agccctggga tggactagat gagcacagcc aagccctgag tgggaggctg
     3781 cgggccattc tccagaatca gggaaacagc ggcagcgaga ctcccgggac ctcagagtcc
     3841 gccacacccg aaagtgataa aaagtattct attggtttag ccatcggcac taattccgtt
     3901 ggatgggctg tcataaccga tgaatacaaa gtaccttcaa agaaatttaa ggtgttgggg
     3961 aacacagacc gtcattcgat taaaaagaat cttatcggtg ccctcctatt cgatagtggc
     4021 gaaacggcag aggcgactcg cctgaaacga accgctcgga gaaggtatac acgtcgcaag
     4081 aaccgaatat gttacttaca agaaattttt agcaatgaga tggccaaagt tgacgattct
     4141 ttctttcacc gtttggaaga gtccttcctt gtcgaagagg acaagaaaca tgaacggcac
     4201 cccatctttg gaaacatagt agatgaggtg gcatatcatg aaaagtaccc aacgatttat
     4261 cacctcagaa aaaagctagt tgactcaact gataaagcgg acctgaggtt aatctacttg
     4321 gctcttgccc atatgataaa gttccgtggg cactttctca ttgagggtga tctaaatccg
     4381 gacaactcgg atgtcgacaa actgttcatc cagttagtac aaacctataa tcagttgttt
     4441 gaagagaacc ctataaatgc aagtggcgtg gatgcgaagg ctattcttag cgcccgcctc
     4501 tctaaatccc gacggctaga aaacctgatc gcacaattac ccggagagaa gaaaaatggg
     4561 ttgttcggta accttatagc gctctcacta ggcctgacac caaattttaa gtcgaacttc
     4621 gacttagctg aagatgccaa attgcagctt agtaaggaca cgtacgatga cgatctcgac
     4681 aatctactgg cacaaattgg agatcagtat gcggacttat ttttggctgc caaaaacctt
     4741 agcgatgcaa tcctcctatc tgacatactg agagttaata ctgagattac caaggcgccg
     4801 ttatccgctt caatgatcaa aaggtacgat gaacatcacc aagacttgac acttctcaag
     4861 gccctagtcc gtcagcaact gcctgagaaa tataaggaaa tattctttga tcagtcgaaa
     4921 aacgggtacg caggttatat tgacggcgga gcgagtcaag aggaattcta caagtttatc
     4981 aaacccatat tagagaagat ggatgggacg gaagagttgc ttgtaaaact caatcgcgaa
     5041 gatctactgc gaaagcagcg gactttcgac aacggtagca ttccacatca aatccactta
     5101 ggcgaattgc atgctatact tagaaggcag gaggattttt atccgttcct caaagacaat
     5161 cgtgaaaaga ttgagaaaat cctaaccttt cgcatacctt actatgtggg acccctggcc
     5221 cgagggaact ctcggttcgc atggatgaca agaaagtccg aagaaacgat tactccatgg
     5281 aattttgagg aagttgtcga taaaggtgcg tcagctcaat cgttcatcga gaggatgacc
     5341 aactttgaca agaatttacc gaacgaaaaa gtattgccta agcacagttt actttacgag
     5401 tatttcacag tgtacaatga actcacgaaa gttaagtatg tcactgaggg catgcgtaaa
     5461 cccgcctttc taagcggaga acagaagaaa gcaatagtag atctgttatt caagaccaac
     5521 cgcaaagtga cagttaagca attgaaagag gactacttta agaaaattga atgcttcgat
     5581 tctgtcgaga tctccggggt agaagatcga tttaatgcgt cacttggtac gtatcatgac
     5641 ctcctaaaga taattaaaga taaggacttc ctggataacg aagagaatga agatatctta
     5701 gaagatatag tgttgactct taccctcttt gaagatcggg aaatgattga ggaaagacta
     5761 aaaacatacg ctcacctgtt cgacgataag gttatgaaac agttaaagag gcgtcgctat
     5821 acgggctggg gacgattgtc gcggaaactt atcaacggga taagagacaa gcaaagtggt
     5881 aaaactattc tcgattttct aaagagcgac ggcttcgcca ataggaactt tatgcagctg
     5941 atccatgatg actctttaac cttcaaagag gatatacaaa aggcacaggt ttccggacaa
     6001 ggggactcat tgcacgaaca tattgcgaat cttgctggtt cgccagccat caaaaagggc
     6061 atactccaga cagtcaaagt agtggatgag ctagttaagg tcatgggacg tcacaaaccg
     6121 gaaaacattg taatcgagat ggcacgcgaa aatcaaacga ctcagaaggg gcaaaaaaac
     6181 agtcgagagc ggatgaagag aatagaagag ggtattaaag aactgggcag ccagatctta
     6241 aaggagcatc ctgtggaaaa tacccaattg cagaacgaga aactttacct ctattaccta
     6301 caaaatggaa gggacatgta tgttgatcag gaactggaca taaaccgttt atctgattac
     6361 gacgtcgatc acattgtacc ccaatccttt ttgaaggacg attcaatcga caataaagtg
     6421 cttacacgct cggataagaa ccgagggaaa agtgacaatg ttccaagcga ggaagtcgta
     6481 aagaaaatga agaactattg gcggcagctc ctaaatgcga aactgataac gcaaagaaag
     6541 ttcgataact taactaaagc tgagaggggt ggcttgtctg aacttgacaa ggccggattt
     6601 attaaacgtc agctcgtgga aacccgccaa atcacaaagc atgttgcaca gatactagat
     6661 tcccgaatga atacgaaata cgacgagaac gataagctga ttcgggaagt caaagtaatc
     6721 actttaaagt caaaattggt gtcggacttc agaaaggatt ttcaattcta taaagttagg
     6781 gagataaata actaccacca tgcgcacgac gcttatctta atgccgtcgt agggaccgca
     6841 ctcattaaga aatacccgaa gctagaaagt gagtttgtgt atggtgatta caaagtttat
     6901 gacgtccgta agatgatcgc gaaaagcgaa caggagatag gcaaggctac agccaaatac
     6961 ttcttttatt ctaacattat gaatttcttt aagacggaaa tcactctggc aaacggagag
     7021 atacgcaaac gacctttaat tgaaaccaat ggggagacag gtgaaatcgt atgggataag
     7081 ggccgggact tcgcgacggt gagaaaagtt ttgtccatgc cccaagtcaa catagtaaag
     7141 aaaactgagg tgcagaccgg agggttttca aaggaatcga ttcttccaaa aaggaatagt
     7201 gataagctca tcgctcgtaa aaaggactgg gacccgaaaa agtacggtgg cttcgatagc
     7261 cctacagttg cctattctgt cctagtagtg gcaaaagttg agaagggaaa atccaagaaa
     7321 ctgaagtcag tcaaagaatt attggggata acgattatgg agcgctcgtc ttttgaaaag
     7381 aaccccatcg acttccttga ggcgaaaggt tacaaggaag taaaaaagga tctcataatt
     7441 aaactaccaa agtatagtct gtttgagtta gaaaatggcc gaaaacggat gttggctagc
     7501 gccggagagc ttcaaaaggg gaacgaactc gcactaccgt ctaaatacgt gaatttcctg
     7561 tatttagcgt cccattacga gaagttgaaa ggttcacctg aagataacga acagaagcaa
     7621 ctttttgttg agcagcacaa acattatctc gacgaaatca tagagcaaat ttcggaattc
     7681 agtaagagag tcatcctagc tgatgccaat ctggacaaag tattaagcgc atacaacaag
     7741 cacagggata aacccatacg tgagcaggcg gaaaatatta tccatttgtt tactcttacc
     7801 aacctcggcg ctccagccgc attcaagtat tttgacacaa cgatagatcg caaacgatac
     7861 acttctacca aggaggtgct agacgcgaca ctgattcacc aatccatcac gggattatat
     7921 gaaactcgga tagatttgtc acagcttggg ggtgactctg gtggttctac taatctgtca
     7981 gatattattg aaaaggagac cggtaagcaa ctggttatcc aggaatccat cctcatgctc
     8041 ccagaggagg tggaagaagt cattgggaac aagccggaaa gcgatatact cgtgcacacc
     8101 gcctacgacg agagcaccga cgagaatgtc atgcttctga ctagcgacgc ccctgaatac
     8161 aagccttggg ctctggtcat acaggatagc aacggtgaga acaagattaa gatgctctct
     8221 ggtggttctc ccaagaagaa gaggaaagtc ggaggcggtg gaagcggcgc aacaaacttc
     8281 tctctgctga aacaagccgg agatgtcgaa gagaatcctg gaccgatggt gagcaagggc
     8341 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     8401 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     8461 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     8521 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     8581 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     8641 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     8701 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     8761 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     8821 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     8881 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     8941 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     9001 accgccgccg ggatcactct cggcatggac gagctgtaca agtgaagcgg ccgcactcct
     9061 caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa
     9121 taccactgag atctttttcc ctctgccaaa aattatgggg acatcatgaa gccccttgag
     9181 catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt
     9241 ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca tcagaatgag
     9301 tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa caaaggttgg
     9361 ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga
     9421 aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta
     9481 acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta
     9541 ctcccagtca tagctgtccc tcttctctta tggagatccc tcgacctgca gcccaagctt
     9601 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     9661 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     9721 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg
     9781 gatccgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     9841 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     9901 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     9961 ggaggcctag gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat
    10021 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
    10081 gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgctgcatt aatgaatcgg
    10141 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
    10201 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
    10261 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
    10321 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
    10381 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
    10441 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
    10501 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
    10561 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
    10621 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaac
//
