LOCUS       sequence-251693-       10678 bp    DNA     circular SYN 14-APR-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 Apr 14, 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"
     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..2449)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(2462..2710)
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             complement(join(2723..2815,2816..2818,2819..2823,2824,
                     2825..3421,3422..3424,3425..6823))
                     /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             complement(join(2723..6796,6797..6799,6800..6823))
                     /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             complement(6872..7468)
                     /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"
     regulatory      7465..7474
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(7540..7559)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          7567..8583
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(7621..7643)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(8584..8861)
                     /label=chicken beta-actin promoter
     enhancer        8863..9242
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9273..9377
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(9378..9446,9447..10238)
                     /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(9596..9615)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(10409..10678,1..319)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
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 ccgcctccga ctttcctctt cttcttggga gaaccaccag
     2461 agagcatctt aatcttgttc tcaccgttgc tatcctgtat gaccagagcc caaggcttgt
     2521 attcaggggc gtcgctagtc agaagcatga cattctcgtc ggtgctctcg tcgtaggcgg
     2581 tgtgcacgag tatatcgctt tccggcttgt tcccaatgac ttcttccacc tcctctggga
     2641 gcatgaggat ggattcctgg ataaccagtt gcttaccggt ctccttttca ataatatctg
     2701 acagattagt agaaccacca gagtcacccc caagctgtga caaatctatc cgagtttcat
     2761 ataatcccgt gatggattgg tgaatcagtg tcgcgtctag cacctccttg gtagaagtgt
     2821 atcgtttgcg atctatcgtt gtgtcaaaat acttgaatgc ggctggagcg ccgaggttgg
     2881 taagagtaaa caaatggata atattttccg cctgctcacg tatgggttta tccctgtgct
     2941 tgttgtatgc gcttaatact ttgtccagat tggcatcagc taggatgact ctcttactga
     3001 attccgaaat ttgctctatg atttcgtcga gataatgttt gtgctgctca acaaaaagtt
     3061 gcttctgttc gttatcttca ggtgaacctt tcaacttctc gtaatgggac gctaaataca
     3121 ggaaattcac gtatttagac ggtagtgcga gttcgttccc cttttgaagc tctccggcgc
     3181 tagccaacat ccgttttcgg ccattttcta actcaaacag actatacttt ggtagtttaa
     3241 ttatgagatc cttttttact tccttgtaac ctttcgcctc aaggaagtcg atggggttct
     3301 tttcaaaaga cgagcgctcc ataatcgtta tccccaataa ttctttgact gacttcagtt
     3361 tcttggattt tcccttctca acttttgcca ctactaggac agaataggca actgtagggc
     3421 tatcgaagcc accgtacttt ttcgggtccc agtccttttt acgagcgatg agcttatcac
     3481 tattcctttt tggaagaatc gattcctttg aaaaccctcc ggtctgcacc tcagttttct
     3541 ttactatgtt gacttggggc atggacaaaa cttttctcac cgtcgcgaag tcccggccct
     3601 tatcccatac gatttcacct gtctccccat tggtttcaat taaaggtcgt ttgcgtatct
     3661 ctccgtttgc cagagtgatt tccgtcttaa agaaattcat aatgttagaa taaaagaagt
     3721 atttggctgt agccttgcct atctcctgtt cgcttttcgc gatcatctta cggacgtcat
     3781 aaactttgta atcaccatac acaaactcac tttctagctt cgggtatttc ttaatgagtg
     3841 cggtccctac gacggcatta agataagcgt cgtgcgcatg gtggtagtta tttatctccc
     3901 taactttata gaattgaaaa tcctttctga agtccgacac caattttgac tttaaagtga
     3961 ttactttgac ttcccgaatc agcttatcgt tctcgtcgta tttcgtattc attcgggaat
     4021 ctagtatctg tgcaacatgc tttgtgattt ggcgggtttc cacgagctga cgtttaataa
     4081 atccggcctt gtcaagttca gacaagccac ccctctcagc tttagttaag ttatcgaact
     4141 ttctttgcgt tatcagtttc gcatttagga gctgccgcca atagttcttc attttcttta
     4201 cgacttcctc gcttggaaca ttgtcacttt tccctcggtt cttatccgag cgtgtaagca
     4261 ctttattgtc gattgaatcg tccttcaaaa aggattgggg tacaatgtga tcgacgtcgt
     4321 aatcagataa acggtttatg tccagttcct gatcaacata catgtccctt ccattttgta
     4381 ggtaatagag gtaaagtttc tcgttctgca attgggtatt ttccacagga tgctccttta
     4441 agatctggct gcccagttct ttaataccct cttctattct cttcatccgc tctcgactgt
     4501 ttttttgccc cttctgagtc gtttgatttt cgcgtgccat ctcgattaca atgttttccg
     4561 gtttgtgacg tcccatgacc ttaactagct catccactac tttgactgtc tggagtatgc
     4621 cctttttgat ggctggcgaa ccagcaagat tcgcaatatg ttcgtgcaat gagtcccctt
     4681 gtccggaaac ctgtgccttt tgtatatcct ctttgaaggt taaagagtca tcatggatca
     4741 gctgcataaa gttcctattg gcgaagccgt cgctctttag aaaatcgaga atagttttac
     4801 cactttgctt gtctcttatc ccgttgataa gtttccgcga caatcgtccc cagcccgtat
     4861 agcgacgcct ctttaactgt ttcataacct tatcgtcgaa caggtgagcg tatgttttta
     4921 gtctttcctc aatcatttcc cgatcttcaa agagggtaag agtcaacact atatcttcta
     4981 agatatcttc attctcttcg ttatccagga agtccttatc tttaattatc tttaggaggt
     5041 catgatacgt accaagtgac gcattaaatc gatcttctac cccggagatc tcgacagaat
     5101 cgaagcattc aattttctta aagtagtcct ctttcaattg cttaactgtc actttgcggt
     5161 tggtcttgaa taacagatct actattgctt tcttctgttc tccgcttaga aaggcgggtt
     5221 tacgcatgcc ctcagtgaca tacttaactt tcgtgagttc attgtacact gtgaaatact
     5281 cgtaaagtaa actgtgctta ggcaatactt tttcgttcgg taaattcttg tcaaagttgg
     5341 tcatcctctc gatgaacgat tgagctgacg cacctttatc gacaacttcc tcaaaattcc
     5401 atggagtaat cgtttcttcg gactttcttg tcatccatgc gaaccgagag ttccctcggg
     5461 ccaggggtcc cacatagtaa ggtatgcgaa aggttaggat tttctcaatc ttttcacgat
     5521 tgtctttgag gaacggataa aaatcctcct gccttctaag tatagcatgc aattcgccta
     5581 agtggatttg atgtggaatg ctaccgttgt cgaaagtccg ctgctttcgc agtagatctt
     5641 cgcgattgag ttttacaagc aactcttccg tcccatccat cttctctaat atgggtttga
     5701 taaacttgta gaattcctct tgactcgctc cgccgtcaat ataacctgcg tacccgtttt
     5761 tcgactgatc aaagaatatt tccttatatt tctcaggcag ttgctgacgg actagggcct
     5821 tgagaagtgt caagtcttgg tgatgttcat cgtacctttt gatcattgaa gcggataacg
     5881 gcgccttggt aatctcagta ttaactctca gtatgtcaga taggaggatt gcatcgctaa
     5941 ggtttttggc agccaaaaat aagtccgcat actgatctcc aatttgtgcc agtagattgt
     6001 cgagatcgtc atcgtacgtg tccttactaa gctgcaattt ggcatcttca gctaagtcga
     6061 agttcgactt aaaatttggt gtcaggccta gtgagagcgc tataaggtta ccgaacaacc
     6121 catttttctt ctctccgggt aattgtgcga tcaggttttc tagccgtcgg gatttagaga
     6181 ggcgggcgct aagaatagcc ttcgcatcca cgccacttgc atttataggg ttctcttcaa
     6241 acaactgatt ataggtttgt actaactgga tgaacagttt gtcgacatcc gagttgtccg
     6301 gatttagatc accctcaatg agaaagtgcc cacggaactt tatcatatgg gcaagagcca
     6361 agtagattaa cctcaggtcc gctttatcag ttgagtcaac tagctttttt ctgaggtgat
     6421 aaatcgttgg gtacttttca tgatatgcca cctcatctac tatgtttcca aagatggggt
     6481 gccgttcatg tttcttgtcc tcttcgacaa ggaaggactc ttccaaacgg tgaaagaaag
     6541 aatcgtcaac tttggccatc tcattgctaa aaatttcttg taagtaacat attcggttct
     6601 tgcgacgtgt ataccttctc cgagcggttc gtttcaggcg agtcgcctct gccgtttcgc
     6661 cactatcgaa taggagggca ccgataagat tctttttaat cgaatgacgg tctgtgttcc
     6721 ccaacacctt aaatttcttt gaaggtactt tgtattcatc ggttatgaca gcccatccaa
     6781 cggaattagt gccgatggct aaaccaatag aatacttttt atcactttcg ggtgtggcgg
     6841 actctgaggt cccgggagtc tcgctgccgc tgtttccctg attctggaga atggcccgca
     6901 gcctcccact cagggcttgg ctgtgctcat ctagtccatc ccagggctgg aagggacatc
     6961 cctggtggtc cacaaaggtg tcccagcagt gcttaaattc atcgtaggtc atgatggaga
     7021 cttgggcccc agcatcccgc agcatttgca gtgcctcctt atataggggg tcgtaatcat
     7081 agatgcgggc agcgaagata cgcagtctca cgtgtgtgtt ctcctgaagg aacgcacgca
     7141 cttccccggc acagccccag gagaagcagg ggctccagga gatgaaccaa gtgaccctgt
     7201 agatctgggc cgggtccaac tgcaaagaag gaaccaggtc caagaagcgc agctccgcat
     7261 ggcggccgta aaagccacag agaagattct tagcctggtt gtgtagaaag cccctgtgct
     7321 ggtccatctt gaccgaggtg ccattgtcca ggcgctccac ttcgtagcac aggtaggtct
     7381 tatgccttcc aatgccattg ttaaagttgg aagtgaatat gtgtggatcc atcaagtgtc
     7441 tgggcccgga tgctgggctg gcttccatgg tggcgaccgg tggatcccgg gcccgcggta
     7501 ccgtcgactg cagaattctt tgccaaaatg atgagacagc acaataacca gcacgttgcc
     7561 caggagctgt aggaaaaaga agaaggcatg aacatggtta gcagaggctc tagagccgcc
     7621 ggtcacacgc cagaagccga accccgccct gccccgtccc ccccgaaggc agccgtcccc
     7681 ccgcggacag ccccgaggct ggagagggag aaggggacgg cggcgcggcg acgcacgaag
     7741 gccctccccg cccatttcct tcctgccggc gccgcaccgc ttcgccccgc gcccgctaga
     7801 gggggtgcgg cggcgcctcc cagatttcgg ctccgcacag atttgggaca aaggaagtcc
     7861 ctgcgccctc tcgcacgatt accataaaag gcaatggctg cggctcgccg cgcctcgaca
     7921 gccgccggcg ctccgggggc cgccgcgccc ctcccccgag ccctccccgg cccgaggcgg
     7981 ccccgccccg cccggcaccc ccacctgccg ccaccccccg cccggcacgg cgagccccgc
     8041 gccacgcccc gtacggagcc ccgcacccga agccgggccg tgctcagcaa ctcggggagg
     8101 ggggtgcagg gggggttgca gcccgaccga cgcgcccaca ccccctgctc acccccccac
     8161 gcacacaccc cgcacgcagc ctttgttccc ctcgcagccc cccccgcacc gcggggcacc
     8221 gcccccggcc gcgctcccct cgcgcacact gcggagcgca caaagccccg cgccgcgccc
     8281 gcagcgctca cagccgccgg gcagcgcgga gccgcacgcg gcgctcccca cgcacacaca
     8341 cacgcacgca ccccccgagc cgctcccccc gcacaaaggg ccctcccgga gcccctcaag
     8401 gctttcacgc agccacagaa aagaaacaag ccgtcattaa accaagcgct aattacagcc
     8461 cggaggagaa gggccgtccc gcccgctcac ctgtgggagt aacgcggtca gtcagagccg
     8521 gggcgggcgg cgcgaggcgg cggcggagcg gggcacgggg cgaaggcagc gcgcagcgac
     8581 tcccgcccgc cgcgcgcttc gctttttata gggccgccgc cgccgccgcc tcgccataaa
     8641 aggaaacttt cggagcgcgc cgctctgatt ggctgccgcc gcacctctcc gcctcgcccc
     8701 gccccgcccc tcgccccgcc ccgccccgcc tggcgcgcgc cccccccccc cccccgcccc
     8761 catcgctgca caaaataatt aaaaaataaa taaatacaaa attgggggtg gggagggggg
     8821 ggagatgggg agagtgaagc agaacgtggg gctcacctcg accatggtaa tagcgatgac
     8881 taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg
     8941 ccaggcgggc catttaccgt cattgacgtc aatagggggc gtacttggca tatgatacac
     9001 ttgatgtact gccaagtggg cagtttaccg taaatactcc acccattgac gtcaatggaa
     9061 agtccctatt ggcgttacta tgggaacata cgtcattatt gacgtcaatg ggcgggggtc
     9121 gttgggcggt cagccaggcg ggccatttac cgtaagttat gtaacgcgga actccatata
     9181 tgggctatga actaatgacc ccgtaattga ttactattaa taactagtca ataatcaatg
     9241 tcgacccagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     9301 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     9361 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     9421 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     9481 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     9541 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     9601 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     9661 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     9721 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     9781 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     9841 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     9901 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     9961 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
    10021 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
    10081 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
    10141 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
    10201 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
    10261 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
    10321 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
    10381 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
    10441 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
    10501 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
    10561 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
    10621 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgg
//
