LOCUS       Exported               11037 bp ds-DNA     circular SYN 21-MAR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pX461-evoCDA1-nSpCas9-empty
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11037)
  AUTHORS   NIH-NCI
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Mar 25, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..11037
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     1..19
                     /label=A192-px459v2-deltaSpCas9-FP
     primer_bind     1..19
                     /label=A219-px459v2-dSpCas9-XhoI-F
     CDS             16..69
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             70..669
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(70..89)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     566..586
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    668..669
                     /label=Sanger sequencing confirmed silent mutation
     primer_bind     complement(697..714)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    703..910
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   919..1059
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   919..1048
                     /label=AAV2 ITR
     rep_origin      1134..1589
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1221..1240)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1431..1452
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1606..1625)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1725..1747
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1785..1803)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1845..1862)
                     /label=pQE promoter
     promoter        1871..1975
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1976..2836
                     /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(2194..2213)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3007..3595
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3228..3247
                     /label=RepO-R
     primer_bind     complement(3354..3372)
                     /label=RepO-F
     primer_bind     3496..3515
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        3657..3897
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     3657..3677
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3701..3725
                     /label=TRC-F
     primer_bind     3828..3847
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        3924..3999
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        4096..4381
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        4383..4660
                     /label=chicken beta-actin promoter
     intron          4661..4888
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      4900..4909
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             4909..4974
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     4962..4998
                     /label=Primer 3
     primer_bind     4962..4998
                     /label=A175-px461-evoCDA1-SpCas9-FP
     primer_bind     complement(4962..4980)
                     /label=A220-px459v2-dSpCas9-XhoI-R
     primer_bind     complement(4962..4980)
                     /label=A193-px459v2-deltaSpCas9-RP
     CDS             4962..4974
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DDDK"
     primer_bind     4978..4997
                     /label=EDb 1.4-CBE_F
     primer_bind     4978..4997
                     /label=EDb 4.4L-CBE_F
     misc_feature    4981..5601
                     /label=evoCDA1
     primer_bind     5092..5138
                     /label=Fragment 1.FOR
     primer_bind     5495..5516
                     /label=Fragment 2.FOR
     primer_bind     complement(5495..5514)
                     /label=Fragment 1.REV
     CDS             5512..5514
                     /codon_start=1
                     /label=lys codon mutated
                     /translation="K"
     primer_bind     complement(5588..5611)
                     /label=EDb 1.4-CBE_R
     primer_bind     complement(5588..5611)
                     /label=EDb 4.4L-CBE_R
     primer_bind     5592..5620
                     /label=EDb 1.4-DBD_F
     primer_bind     5592..5620
                     /label=EDb 4.4L-DBD_F
     primer_bind     5595..5632
                     /label=EDb 4.0L (not 4.-1L)-DBD_F
     primer_bind     5595..5626
                     /label=EDb 1.0 (not 1.-1)-DBD_F
     CDS             5601
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation=""
     misc_feature    5602..5697
                     /label=Linker
     primer_bind     5602..5620
                     /label=EDb 1.3-DBD_F
     primer_bind     5602..5620
                     /label=EDb 4.3L-DBD_F
     misc_feature    5698..6039
                     /label=ss DNA-binding domain
     primer_bind     5782..5802
                     /label=ssDBD seq F
     CDS             6040..6135
                     /codon_start=1
                     /label=Linker
                     /translation="SGGSSGGSSGSETPGTSESATPESSGGSSGGS"
     primer_bind     complement(6108..6153)
                     /label=EDb 1.0 (not 1.-1)-DBD_R
     primer_bind     complement(6111..6129)
                     /label=EDb 4.0L (not 4.-1L)-DBD_R
     primer_bind     complement(6115..6153)
                     /label=EDb 1.1_R
     primer_bind     complement(6115..6153)
                     /label=EDb 1.2_R
     primer_bind     complement(6115..6141)
                     /label=EDb 4.3L-DBD_R
     primer_bind     complement(6115..6141)
                     /label=EDb 4.4L-DBD_R
     primer_bind     complement(6115..6141)
                     /label=EDb 4.1L_R
     primer_bind     complement(6117..6163)
                     /label=Fragment 2.REV
     primer_bind     complement(6119..6158)
                     /label=EDb 1.3-DBD_R
     primer_bind     complement(6119..6158)
                     /label=EDb 1.4-DBD_R
     primer_bind     complement(6122..6141)
                     /label=EDb 4.2L_R
     primer_bind     6136..6163
                     /label=EDb 1.0/3.0_F
     CDS             6142..10242
                     /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"
     primer_bind     complement(6197..6217)
                     /label=Cas9 rev seq1
     primer_bind     6510..6529
                     /label=Cas9_confirm_2
     primer_bind     7110..7129
                     /label=Cas9_confirm_3
     primer_bind     7710..7729
                     /label=Cas9_confirm_4
     primer_bind     8290..8309
                     /label=Cas9_5_F
     primer_bind     8290..8309
                     /label=Cas9_5_F (1)
     primer_bind     8310..8325
                     /label=Cas9_confirm_5
     primer_bind     8493..8512
                     /label=Cas9 intern seq 3
     primer_bind     8910..8929
                     /label=Cas9_confirm_6
     primer_bind     9510..9532
                     /label=Cas9_confirm_7
     primer_bind     complement(9745..9765)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     10110..10134
                     /label=Cas9_comfirm_8
     primer_bind     complement(10230..10266)
                     /label=EDb 1.-1-Cas9_R
     primer_bind     complement(10238..10250)
                     /label=ED 3.0_R
     primer_bind     10247..10293
                     /label=EDb 1.-1/4.-1-UGI_F
     primer_bind     10273..10293
                     /label=EDb 1.0/4.0-UGI_F
     CDS             10279..10527
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             10558..10806
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     primer_bind     complement(10803..10842)
                     /label=Fragment.REV
     primer_bind     complement(10803..10840)
                     /label=EDb 1.0-UGI_R
     primer_bind     complement(10803..10818)
                     /label=Infusion fragment R
     primer_bind     10824..10848
                     /label=inf 6nls.FOR
     primer_bind     complement(10826..10842)
                     /label=Infusion vec R
     primer_bind     complement(10826..10840)
                     /label=ED 3.0_R
     CDS             10831..10857
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             10867..10893
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             10903..10929
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             10939..10965
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             10975..11001
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             11011..11037
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     primer_bind     complement(join(11015..11037,1..19))
                     /label=Primer 4
     primer_bind     complement(join(11015..11037,1..19))
                     /label=A179-px461-evoCDA1-SpCas9-RP
     primer_bind     complement(11017..11037)
                     /label=EDi 1-3_R
     primer_bind     complement(join(11019..11037,1))
                     /label=inf 6nls.REV
ORIGIN
        1 gaattcggca gtggagaggg cagaggaagt ctgctaacat gcggtgacgt cgaggagaat
       61 cctggcccaa tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
      121 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
      181 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
      241 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
      301 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
      361 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
      421 cccaaggagc ccgcgtggtt cctggccacc gtcggagtct cgcccgacca ccagggcaag
      481 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
      541 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
      601 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
      661 ggtgcctagg aattctaact agagctcgct gatcagcctc gactgtgcct tctagttgcc
      721 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
      781 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
      841 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagag aatagcaggc
      901 atgctgggga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
      961 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     1021 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     1081 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     1141 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     1201 tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     1261 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     1321 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     1381 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     1441 gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat
     1501 tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     1561 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     1621 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     1681 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     1741 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     1801 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     1861 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     1921 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     1981 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     2041 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     2101 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     2161 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     2221 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     2281 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     2341 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     2401 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     2461 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     2521 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     2581 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     2641 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg gaagccgcgg
     2701 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     2761 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     2821 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     2881 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     2941 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3001 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3061 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3121 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3181 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3241 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3301 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3361 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3421 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3481 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3541 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3601 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtgagg
     3661 gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt agagagataa
     3721 ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt
     3781 aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc
     3841 ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa
     3901 acaccgggtc ttcgagaaga cctgttttag agctagaaat agcaagttaa aataaggcta
     3961 gtccgttatc aacttgaaaa agtggcaccg agtcggtgct tttttgtttt agagctagaa
     4021 atagcaagtt aaaataaggc tagtccgttt ttagcgcgtg cgccaattct gcagacaaat
     4081 ggctctagag gtacccgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     4141 cgacccccgc ccattgacgt caatagtaac gccaataggg actttccatt gacgtcaatg
     4201 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     4261 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattgtg cccagtacat
     4321 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     4381 ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc
     4441 aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggggg
     4501 ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg ggcggggcga ggcggagagg
     4561 tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg
     4621 gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct
     4681 tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc
     4741 gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagctgag
     4801 caagaggtaa gggtttaagg gatggttggt tggtggggta ttaatgttta attacctgga
     4861 gcacctgcct gaaatcactt tttttcaggt tggaccggtg ccaccatgga ctataaggac
     4921 cacgacggag actacaagga tcatgatatt gattacaaag acgatgacga taagatggcc
     4981 accgacgccg agtacgtgcg gatccacgag aagctggata tctatacatt caagaagcag
     5041 tttagcaaca ataagaagtc cgtgtctcac agatgctacg tgctgttcga gctgaagcgg
     5101 agaggagaga ggcgcgcctg tttttggggc tatgccgtga acaagccaca gtctggaacc
     5161 gagaggggaa tccacgcaga gatcttcagc atcaggaagg tggaggagta cctgcgcgac
     5221 aaccccggcc agtttacaat caattggtat agctcctgga gcccttgcgc cgattgtgcc
     5281 gagaagatcc tggagtggta caaccaggag ctgaggggca atggccacac cctgaagatc
     5341 tgggtgtgca agctgtacta tgagaagaac gccaggaatc agatcggcct gtggaacctg
     5401 cgcgacaatg gcgtgggcct gaacgtgatg gtgtccgagc actatcagtg ctgtcgcaag
     5461 atctttatcc agtctagcca caatcagctg aacgagaatc ggtggctgga gaaaacactg
     5521 aagagagccg agaagcggag aagcgagctg tccatcatgt ttcaggtgaa gatcctgcac
     5581 accacaaagt ctcccgccgt gagcggagga tctagcggag gatcaagcgg aagcgagact
     5641 cctggaacca gcgaaagcgc aaccccagaa agcagcggag gaagtagcgg aggaagcatg
     5701 gcaatgcaga tgcagcttga agcaaatgca gatacttcag tggaagaaga aagctttggc
     5761 ccacaaccca tttcacggtt agagcagtgt ggcataaatg ccaacgatgt gaagaaattg
     5821 gaagaagctg gattccatac tgtggaggct gttgcctatg cgccaaagaa ggagctaata
     5881 aatattaagg gaattagtga agccaaagct gataaaattc tggctgaggc agctaaatta
     5941 gttccaatgg gtttcaccac tgcaactgaa ttccaccaaa ggcggtcaga gatcatacag
     6001 attactactg gctccaaaga gcttgacaaa ctacttcaat ctggcggatc tagcggtgga
     6061 tctagcggct ctgagacccc tggaacatcc gaatccgcca ctccagagag cagcggaggc
     6121 tcttctggag gatcaagcgc tgacaagaag tacagcatcg gcctggccat cggcaccaac
     6181 tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg
     6241 ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggagccct gctgttcgac
     6301 agcggcgaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga
     6361 cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac
     6421 gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag
     6481 cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc
     6541 atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc
     6601 tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg
     6661 aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag
     6721 ctgttcgagg aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc
     6781 agactgagca agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag
     6841 aatggcctgt tcggaaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc
     6901 aacttcgacc tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac
     6961 ctggacaacc tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag
     7021 aacctgtccg acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag
     7081 gcccccctga gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg
     7141 ctgaaagctc tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag
     7201 agcaagaacg gctacgccgg ctacattgac ggcggagcca gccaggaaga gttctacaag
     7261 ttcatcaagc ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac
     7321 agagaggacc tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc
     7381 cacctgggag agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag
     7441 gacaaccggg aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct
     7501 ctggccaggg gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc
     7561 ccctggaact tcgaggaagt ggtggacaag ggcgcttccg cccagagctt catcgagcgg
     7621 atgaccaact tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg
     7681 tacgagtact tcaccgtgta taacgagctg accaaagtga aatacgtgac cgagggaatg
     7741 agaaagcccg ccttcctgag cggcgagcag aaaaaggcca tcgtggacct gctgttcaag
     7801 accaaccgga aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc
     7861 ttcgactccg tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac
     7921 cacgatctgc tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac
     7981 attctggaag atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa
     8041 cggctgaaaa cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg
     8101 agatacaccg gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag
     8161 tccggcaaga caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg
     8221 cagctgatcc acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc
     8281 ggccagggcg atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag
     8341 aagggcatcc tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac
     8401 aagcccgaga acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag
     8461 aagaacagcc gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag
     8521 atcctgaaag aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac
     8581 tacctgcaga atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc
     8641 gactacgatg tggaccatat cgtgcctcag agctttctga aggacgactc catcgacaac
     8701 aaggtgctga ccagaagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag
     8761 gtcgtgaaga agatgaagaa ctactggcgg cagctgctga acgccaagct gattacccag
     8821 agaaagttcg acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc
     8881 ggcttcatca agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc
     8941 ctggactccc ggatgaacac taagtacgac gagaatgaca agctgatccg ggaagtgaaa
     9001 gtgatcaccc tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa
     9061 gtgcgcgaga tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga
     9121 accgccctga tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag
     9181 gtgtacgacg tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc
     9241 aagtacttct tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac
     9301 ggcgagatcc ggaagcggcc tctgatcgag acaaacggcg aaaccgggga gatcgtgtgg
     9361 gataagggcc gggattttgc caccgtgcgg aaagtgctga gcatgcccca agtgaatatc
     9421 gtgaaaaaga ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg
     9481 aacagcgata agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc
     9541 gacagcccca ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc
     9601 aagaaactga agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc
     9661 gagaagaatc ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg
     9721 atcatcaagc tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg
     9781 gcctctgccg gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac
     9841 ttcctgtacc tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag
     9901 aaacagctgt ttgtggaaca gcacaagcac tacctggacg agatcatcga gcagatcagc
     9961 gagttctcca agagagtgat cctggccgac gctaatctgg acaaagtgct gtccgcctac
    10021 aacaagcacc gggataagcc catcagagag caggccgaga atatcatcca cctgtttacc
    10081 ctgaccaatc tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag
    10141 aggtacacca gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc
    10201 ctgtacgaga cacggatcga cctgtctcag ctgggaggtg acagtggcgg aagtggacta
    10261 gtaggaagtg gcgggagcac taatctgagc gacatcattg agaaggagac tgggaaacag
    10321 ctggtcattc aggagtccat cctgatgctg cctgaggagg tggaggaagt gatcggcaac
    10381 aagccagagt ctgacatcct ggtgcacacc gcctacgacg agtccacaga tgagaatgtg
    10441 atgctgctga cctctgacgc ccccgagtat aagccttggg ccctggtcat ccaggattct
    10501 aacggcgaga ataagatcaa gatgctgagc ggaggatccg gaggatctgg aggcagcacc
    10561 aacctgtctg acatcatcga gaaggagaca ggcaagcagc tggtcatcca ggagagcatc
    10621 ctgatgctgc ccgaagaagt cgaagaagtg atcggaaaca agcctgagag cgatatcctg
    10681 gtccataccg cctacgacga gagtaccgac gaaaatgtga tgctgctgac atccgacgcc
    10741 ccagagtata agccctgggc tctggtcatc caggattcca acggagagaa caaaatcaaa
    10801 atgctgtctg gcggctcaga attcggatcc cctgctgcta aacgtgttaa gcttgatggg
    10861 ggtagcccgg cagccaagag agtcaaactc gacggcggct ccccagcggc aaaaagggtg
    10921 aaactagacg ggggtagccc cgccgcgaag cgtgtaaagc tggatggagg atcgcctgcg
    10981 gctaagcgag tcaaattaga cggtgggtca ccggctgcaa agcgcgtcaa attggat
//
