pX330a dCas9-LSD1

LOCUS       pX330a dCas9-LSD1              10984 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Modified CAG promoter-containing vector for ubiquitous expression of
            catalytically inactive Cas9 fused to LSD1. For targeted enhancer 
            demethylation in chicken embryos..
ACCESSION   .
VERSION     .
KEYWORDS    pX330a dCas9-LSD1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10984)
  AUTHORS   Williams RM, Senanayake U, Artibani M, Taylor G, Wells D, Ahmed AA, 
            Sauka-Spengler T
  TITLE     Genome and epigenome engineering CRISPR toolkit for in vivo 
            modulation of cis-regulatory interactions and gene expression in the
            chicken embryo.
  JOURNAL   Development. 2018 Feb 23;145(4). pii: dev.160333. doi: 
            10.1242/dev.160333.
  PUBMED    29386245
REFERENCE   2  (bases 1 to 10984)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10984
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      24..612
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     513..532
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        674..914
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     674..694
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     845..864
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        941..1016
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        1113..1398
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1400..1677
                     /label=chicken beta-actin promoter
     intron          1678..1906
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             1927..1947
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1957..6060
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=Cas9m4
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the mutations D10A, D839A, 
                     H840A, and N863A (Mali et al., 2013)"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVAAIVPQSFLKDDSIDNKVLTRSDKARGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             6073..6093
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(8698..8715)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8704..8911
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   8920..9060
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   8920..9049
                     /label=AAV2 ITR
     rep_origin      9135..9590
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9222..9241)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9432..9453
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9607..9626)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9726..9748
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(9786..9804)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        9872..9976
                     /gene="bla"
                     /label=AmpR promoter
     CDS             9977..10837
                     /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(10195..10214)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
       61 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
      121 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
      181 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
      241 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
      301 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
      361 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
      421 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
      481 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
      541 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
      601 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
      661 tttgctcaca tgtgagggcc tatttcccat gattccttca tatttgcata tacgatacaa
      721 ggctgttaga gagataattg gaattaattt gactgtaaac acaaagatat tagtacaaaa
      781 tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa
      841 aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat
      901 cttgtggaaa ggacgaaaca ccgggtcttc gagaagacct gttttagagc tagaaatagc
      961 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
     1021 ttgttttaga gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc
     1081 caattctgca gacaaatggc tctagaggta cccgttacat aacttacggt aaatggcccg
     1141 cctggctgac cgcccaacga cccccgccca ttgacgtcaa tagtaacgcc aatagggact
     1201 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     1261 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     1321 cattgtgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     1381 gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc
     1441 cccccctccc cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg
     1501 ggggcggggg gggggggggg gcgcgcgcca ggcggggcgg ggcggggcga ggggcggggc
     1561 ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct
     1621 tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga
     1681 gtcgctgcga cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc
     1741 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
     1801 gggctgtaat tagctgagca agaggtaagg gtttaaggga tggttggttg gtggggtatt
     1861 aatgtttaat tacctggagc acctgcctga aatcactttt tttcaggttg gaccggtgcc
     1921 accatgccca agaagaagag gaaggtggga agggggatgg acaagaagta ctccattggg
     1981 ctcgctatcg gcacaaacag cgtcggctgg gccgtcatta cggacgagta caaggtgccg
     2041 agcaaaaaat tcaaagttct gggcaatacc gatcgccaca gcataaagaa gaacctcatt
     2101 ggcgccctcc tgttcgactc cggggagacg gccgaagcca cgcggctcaa aagaacagca
     2161 cggcgcagat atacccgcag aaagaatcgg atctgctacc tgcaggagat ctttagtaat
     2221 gagatggcta aggtggatga ctctttcttc cataggctgg aggagtcctt tttggtggag
     2281 gaggataaaa agcacgagcg ccacccaatc tttggcaata tcgtggacga ggtggcgtac
     2341 catgaaaagt acccaaccat atatcatctg aggaagaagc ttgtagacag tactgataag
     2401 gctgacttgc ggttgatcta tctcgcgctg gcgcatatga tcaaatttcg gggacacttc
     2461 ctcatcgagg gggacctgaa cccagacaac agcgatgtcg acaaactctt tatccaactg
     2521 gttcagactt acaatcagct tttcgaagag aacccgatca acgcatccgg agttgacgcc
     2581 aaagcaatcc tgagcgctag gctgtccaaa tcccggcggc tcgaaaacct catcgcacag
     2641 ctccctgggg agaagaagaa cggcctgttt ggtaatctta tcgccctgtc actcgggctg
     2701 acccccaact ttaaatctaa cttcgacctg gccgaagatg ccaagcttca actgagcaaa
     2761 gacacctacg atgatgatct cgacaatctg ctggcccaga tcggcgacca gtacgcagac
     2821 ctttttttgg cggcaaagaa cctgtcagac gccattctgc tgagtgatat tctgcgagtg
     2881 aacacggaga tcaccaaagc tccgctgagc gctagtatga tcaagcgcta tgatgagcac
     2941 caccaagact tgactttgct gaaggccctt gtcagacagc aactgcctga gaagtacaag
     3001 gaaattttct tcgatcagtc taaaaatggc tacgccggat acattgacgg cggagcaagc
     3061 caggaggaat tttacaaatt tattaagccc atcttggaaa aaatggacgg caccgaggag
     3121 ctgctggtaa agcttaacag agaagatctg ttgcgcaaac agcgcacttt cgacaatgga
     3181 agcatccccc accagattca cctgggcgaa ctgcacgcta tcctcaggcg gcaagaggat
     3241 ttctacccct ttttgaaaga taacagggaa aagattgaga aaatcctcac atttcggata
     3301 ccctactatg taggccccct cgcccgggga aattccagat tcgcgtggat gactcgcaaa
     3361 tcagaagaga ccatcactcc ctggaacttc gaggaagtcg tggataaggg ggcctctgcc
     3421 cagtccttca tcgaaaggat gactaacttt gataaaaatc tgcctaacga aaaggtgctt
     3481 cctaaacact ctctgctgta cgagtacttc acagtttata acgagctcac caaggtcaaa
     3541 tacgtcacag aagggatgag aaagccagca ttcctgtctg gagagcagaa gaaagctatc
     3601 gtggacctcc tcttcaagac gaaccggaaa gttaccgtga aacagctcaa agaggactat
     3661 ttcaaaaaga ttgaatgttt cgactctgtt gaaatcagcg gagtggagga tcgcttcaac
     3721 gcatccctgg gaacgtatca cgatctcctg aaaatcatta aagacaagga cttcctggac
     3781 aatgaggaga acgaggacat tcttgaggac attgtcctca cccttacgtt gtttgaagat
     3841 agggagatga ttgaagaacg cttgaaaact tacgctcatc tcttcgacga caaagtcatg
     3901 aaacagctca agaggcgccg atatacagga tgggggcggc tgtcaagaaa actgatcaat
     3961 gggatccgag acaagcagag tggaaagaca atcctggatt ttcttaagtc cgatggattt
     4021 gccaaccgga acttcatgca gttgatccat gatgactctc tcacctttaa ggaggacatc
     4081 cagaaagcac aagtttctgg ccagggggac agtctccacg agcacatcgc taatcttgca
     4141 ggtagcccag ctatcaaaaa gggaatactg cagaccgtta aggtcgtgga tgaactcgtc
     4201 aaagtaatgg gaaggcataa gcccgagaat atcgttatcg agatggcccg agagaaccaa
     4261 actacccaga agggacagaa gaacagtagg gaaaggatga agaggattga agagggtata
     4321 aaagaactgg ggtcccaaat ccttaaggaa cacccagttg aaaacaccca gcttcagaat
     4381 gagaagctct acctgtacta cctgcagaac ggcagggaca tgtacgtgga tcaggaactg
     4441 gacatcaatc ggctctccga ctacgacgtg gctgctatcg tgccccagtc ttttctcaaa
     4501 gatgattcta ttgataataa agtgttgaca agatccgata aagctagagg gaagagtgat
     4561 aacgtcccct cagaagaagt tgtcaagaaa atgaaaaatt attggcggca gctgctgaac
     4621 gccaaactga tcacacaacg gaagttcgat aatctgacta aggctgaacg aggtggcctg
     4681 tctgagttgg ataaagccgg cttcatcaaa aggcagcttg ttgagacacg ccagatcacc
     4741 aagcacgtgg cccaaattct cgattcacgc atgaacacca agtacgatga aaatgacaaa
     4801 ctgattcgag aggtgaaagt tattactctg aagtctaagc tggtctcaga tttcagaaag
     4861 gactttcagt tttataaggt gagagagatc aacaattacc accatgcgca tgatgcctac
     4921 ctgaatgcag tggtaggcac tgcacttatc aaaaaatatc ccaagcttga atctgaattt
     4981 gtttacggag actataaagt gtacgatgtt aggaaaatga tcgcaaagtc tgagcaggaa
     5041 ataggcaagg ccaccgctaa gtacttcttt tacagcaata ttatgaattt tttcaagacc
     5101 gagattacac tggccaatgg agagattcgg aagcgaccac ttatcgaaac aaacggagaa
     5161 acaggagaaa tcgtgtggga caagggtagg gatttcgcga cagtccggaa ggtcctgtcc
     5221 atgccgcagg tgaacatcgt taaaaagacc gaagtacaga ccggaggctt ctccaaggaa
     5281 agtatcctcc cgaaaaggaa cagcgacaag ctgatcgcac gcaaaaaaga ttgggacccc
     5341 aagaaatacg gcggattcga ttctcctaca gtcgcttaca gtgtactggt tgtggccaaa
     5401 gtggagaaag ggaagtctaa aaaactcaaa agcgtcaagg aactgctggg catcacaatc
     5461 atggagcgat caagcttcga aaaaaacccc atcgactttc tcgaggcgaa aggatataaa
     5521 gaggtcaaaa aagacctcat cattaagctt cccaagtact ctctctttga gcttgaaaac
     5581 ggccggaaac gaatgctcgc tagtgcgggc gagctgcaga aaggtaacga gctggcactg
     5641 ccctctaaat acgttaattt cttgtatctg gccagccact atgaaaagct caaagggtct
     5701 cccgaagata atgagcagaa gcagctgttc gtggaacaac acaaacacta ccttgatgag
     5761 atcatcgagc aaataagcga attctccaaa agagtgatcc tcgccgacgc taacctcgat
     5821 aaggtgcttt ctgcttacaa taagcacagg gataagccca tcagggagca ggcagaaaac
     5881 attatccact tgtttactct gaccaacttg ggcgcgcctg cagccttcaa gtacttcgac
     5941 accaccatag acagaaagcg gtacacctct acaaaggagg tcctggacgc cacactgatt
     6001 catcagtcaa ttacggggct ctatgaaaca agaatcgacc tctctcagct cggtggagac
     6061 agcagggctg accccaagaa gaagaggaag gtggaggcca gcggaggagg atctttatct
     6121 gggaagaagg cggcagccgc ggcggcggcg gctgcagcgg cagcaaccgg gacggaggct
     6181 ggccctggga cagcaggcgg ctccgagaac gggtctgagg tggccgcgca gcccgcgggc
     6241 ctgtcgggcc cagccgaggt cgggccgggg gcggtggggg agcgcacacc ccgcaagaaa
     6301 gagcctccgc gggcctcgcc ccccgggggc ctggcggaac cgccggggtc cgcagggcct
     6361 caggccggcc ctactgtcgt gcctgggtct gcgaccccca tggaaactgg aatagcagag
     6421 actccggagg ggcgtcggac cagccggcgc aagcgggcga aggtagagta cagagagatg
     6481 gatgaaagct tggccaacct ctcagaagat gagtattatt cagaagaaga gagaaatgcc
     6541 aaagcagaga aggaaaagaa gcttccccca ccaccccctc aagccccacc tgaggaagaa
     6601 aatgaaagtg agcctgaaga accatcgggt gtggagggcg cagctttcca gagccgactt
     6661 cctcatgacc ggatgacttc tcaagaagca gcctgttttc cagatattat cagtggacca
     6721 caacagaccc agaaggtttt tcttttcatt agaaaccgca cactgcagtt gtggttggat
     6781 aatccaaaga ttcagctgac atttgaggct actctccaac aattagaagc accttataac
     6841 agtgatactg tgcttgtcca ccgagttcac agttatttag agcgtcatgg tcttatcaac
     6901 ttcggcatct ataagaggat aaaaccccta ccaactaaaa agacaggaaa ggtaattatt
     6961 ataggctctg gggtctcagg cttggcagca gctcgacagt tacaaagttt tggaatggat
     7021 gtcacacttt tggaagccag ggatcgtgtg ggtggacgag ttgccacatt tcgcaaagga
     7081 aactatgtag ctgatcttgg agccatggtg gtaacaggtc ttggagggaa tcctatggct
     7141 gtggtcagca aacaagtaaa tatggaactg gccaagatca agcaaaaatg cccactttat
     7201 gaagccaacg gacaagctgt tcctaaagag aaagatgaaa tggtagagca agagtttaac
     7261 cggttgctag aagctacatc ttaccttagt catcaactag acttcaatgt cctcaataat
     7321 aagcatgtgt cccttggcca ggcattggaa gttgtcattc agttacaaga gaagcatgtc
     7381 aaagatgagc agattgaaca ttggaagaag atagtgaaaa ctcaggaaga attgaaagaa
     7441 cttcttaata agatggtaaa tttgaaagag aaaattaaag aactccatca gcaatacaaa
     7501 gaagcatctg aagtaaagcc acccagagat attactgccg agttcttagt gaaaagcaaa
     7561 cacagggatc tgaccgccct atgcaaggaa tatgatgaat tagctgaaac acaaggaaag
     7621 ctagaagaaa aacttcagga gttggaagcg aatcccccaa gtgatgtata tctctcatca
     7681 agagacagac aaatacttga ttggcatttt gcaaatcttg aatttgctaa tgccacacct
     7741 ctctcaactc tctcccttaa gcactgggat caggatgatg actttgagtt cactggcagc
     7801 cacctgacag taaggaatgg ctactcgtgt gtgcctgtgg ctttagcaga aggcctagac
     7861 attaaactga atacagcagt gcgacaggtt cgctacacgg cttcaggatg tgaagtgata
     7921 gctgtgaata cccgctccac gagtcaaacc tttatttata aatgcgacgc agttctctgt
     7981 acccttcccc tgggtgtgct gaagcagcag ccaccagccg ttcagtttgt gccacctctc
     8041 cctgagtgga aaacatctgc agtccaaagg atgggatttg gcaaccttaa caaggtggtg
     8101 ttgtgttttg atcgggtgtt ctgggatcca agtgtcaatt tgttcgggca tgttggcagt
     8161 acgactgcca gcaggggtga gctcttcctc ttctggaacc tctataaagc tccaatactg
     8221 ttggcactag tggcaggaga agctgctggt atcatggaaa acataagtga cgatgtgatt
     8281 gttggccgat gcctggccat tctcaaaggg atttttggta gcagtgcagt acctcagccc
     8341 aaagaaactg tggtgtctcg ttggcgtgct gatccctggg ctcggggctc ttattcctat
     8401 gttgctgcag gatcatctgg aaatgactat gatttaatgg ctcagccaat cactcctggc
     8461 ccctcgattc caggtgcccc acagccgatt ccacgactct tctttgcggg agaacatacg
     8521 atccgtaact acccagccac agtgcatggt gctctgctga gtgggctgcg agaagcggga
     8581 agaattgcag accagttttt gggggccatg tatacgctgc ctcgccaggc cacaccaggt
     8641 gttcctgcac agcagtcccc aagcatgtga taagaattcc tagagctcgc tgatcagcct
     8701 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     8761 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     8821 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     8881 attgggaaga gaatagcagg catgctgggg agcggccgca ggaaccccta gtgatggagt
     8941 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     9001 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     9061 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     9121 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     9181 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     9241 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     9301 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     9361 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     9421 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcgggcta
     9481 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     9541 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     9601 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     9661 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     9721 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     9781 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     9841 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     9901 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     9961 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
    10021 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
    10081 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
    10141 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
    10201 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
    10261 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
    10321 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
    10381 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
    10441 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
    10501 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
    10561 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
    10621 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
    10681 cggtgagcgt ggaagccgcg gtatcattgc agcactgggg ccagatggta agccctcccg
    10741 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
    10801 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
    10861 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
    10921 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
    10981 cccc
//