pG3K-U6SC

LOCUS       pG3K-U6SC              13021 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  pGreen3 CRISPR/Cas9 .
ACCESSION   .
VERSION     .
KEYWORDS    pG3K-U6SC
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13021)
  AUTHORS   Zhang Q, Zhang Y, Lu MH, Chai YP, Jiang YY, Zhou Y, Wang XC, Chen QJ
  TITLE     A Novel Ternary Vector System United with Morphogenic Genes Enhances
            CRISPR/Cas Delivery in Maize.
  JOURNAL   Plant Physiol. 2019 Sep 26. pii: pp.19.00767. doi: 
            10.1104/pp.19.00767.
  PUBMED    31558579
REFERENCE   2  (bases 1 to 13021)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..13021
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..25
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     213..235
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     227..244
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     228..244
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        700..804
                     /gene="bla"
                     /label=AmpR promoter
     CDS             805..1665
                     /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(1023..1042)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     misc_RNA        1814..1889
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             2865..2930
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2937..2957
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2982..7082
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     RBS             6995..7003
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             7083..7130
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(7787..7803)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7787..7803)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7800..7822)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7811..7827
                     /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(7835..7865)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7880..7901
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8092..8768
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     8705..8725
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             8831..9628
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
                     SSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
                     QGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
                     ALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(8888..8907)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9498..9517
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    9685..9859
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    9937..9961
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     complement(10339..10358)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             10350..11159
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVSLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(10417..10436)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      11238..11324
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      11416..11443
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      11522..12110
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12011..12030
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtttacccgc caatatatcc tgtcaaacac tgatagttta aactgaaggc gggaaacgac
       61 aatctgatcc aagctcaagc tgctctagca ttcgccattc aggctgcgca actgttggga
      121 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
      181 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
      241 cagtgccaag cttcgacttg ccttccgcac aatacatcat ttcttcttag ctttttttct
      301 tcttcttcgt tcatacagtt tttttttgtt tatcagctta cattttcttg aaccgtagct
      361 ttcgttttct tctttttaac tttccattcg gagtttttgt atcttgtttc atagtttgtc
      421 ccaggattag aatgattagg catcgaacct tcaagaattt gattgaataa aacatcttca
      481 ttcttaagat atgaagataa tcttcaaaag gcccctggga atctgaaaga agagaagcag
      541 gcccatttat atgggaaaga acaatagtat ttcttatata ggcccattta agttgaaaac
      601 aatcttcaaa agtcccacat cgcttagata agaaaacgaa gctgagttta tatacagcta
      661 gagtcgaagt agtgattgtg agaccttcgg ggaaatgtgc gcggaacccc tatttgttta
      721 tttttctaaa tacattcaaa tatgtatccg ctcatgggac aataaccctg ataaatgctt
      781 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
      841 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
      901 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
      961 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     1021 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     1081 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     1141 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     1201 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     1261 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     1321 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     1381 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     1441 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     1501 tctggagccg gtgagcgtgg ctctcgcggt atcattgcag cactggggcc agatggtaag
     1561 ccctcccgta tcgtagttat ctacaccacg gggagtcagg caactatgga tgaacgaaat
     1621 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     1681 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     1741 aagatccttt ttgataatcc catgaccaaa atcccttaac gtgagttttc gttccactga
     1801 gcgtcaggtc tcagttttag agctagaaat agcaagttaa aataaggcta gtccgttatc
     1861 aacttgaaaa agtggcaccg agtcggtgct tttttttgga tagaatttcc cagctttttt
     1921 gcgtgtttca gctctcatga tccttgcatg cactagtatg gagtcaaaga ttcaaataga
     1981 ggacctaaca gaactcgccg taaagactgg cgaacagttc atacagagtc tcttacgact
     2041 caatgacaag aagaaaatct tcgtcaacat ggtggagcac gacacacttg tctactccaa
     2101 aaatatcaaa gatacagtct cagaagacca aagggcaatt gagacttttc aacaaagggt
     2161 aatatccgga aacctcctcg gattccattg cccagctatc tgtcacttta ttgtgaagat
     2221 agtggaaaag gaaggtggct cctacaaatg ccatcattgc gataaaggaa aggccatcgt
     2281 tgaagatgcc tctgccgaca gtggtcccaa agatggaccc ccacccacga ggagcatcgt
     2341 ggaaaaagaa gacgttccaa ccacgtcttc aaagcaagtg gattgatgtg attggcagac
     2401 atactgtccc acaaatgaag atggaatctg taaaagaaaa cgcgtgaaat aatgcgtctg
     2461 acaaaggtta ggtcggctgc ctttaatcaa taccaaagtg gtccctacca cgatggaaaa
     2521 actgtgcagt cggtttggct ttttctgacg aacaaataag attcgtggcc gacaggtggg
     2581 ggtccaccat gtgaaggcat cttcagactc caataatgga gcaatgacgt aagggcttac
     2641 gaaataagta agggtagttt gggaaatgtc cactcacccg tcagtctata aatacttagc
     2701 ccctccctca ttgttaaggg agcaaaatct cagagagata gtcctagaga gagaaagaga
     2761 gcaagtagcc tagaagtagt caaggcggcg aagtattcag gcacgtggcc aggaagaaga
     2821 aaagccaaga cgacgaaaac aggtaagagc taagcatcta gatggattac aaggaccacg
     2881 acggggatta caaggaccac gacattgatt acaaggatga tgatgacaag atggctccga
     2941 agaagaagag gaaggttggc atccacgggg tgccagctgc tgacaagaag tactcgatcg
     3001 gcctcgatat tgggactaac tctgttggct gggccgtgat caccgacgag tacaaggtgc
     3061 cctcaaagaa gttcaaggtc ctgggcaaca ccgatcggca ttccatcaag aagaatctca
     3121 ttggcgctct cctgttcgac agcggcgaga cggctgaggc tacgcggctc aagcgcaccg
     3181 cccgcaggcg gtacacgcgc aggaagaatc gcatctgcta cctgcaggag attttctcca
     3241 acgagatggc gaaggttgac gattctttct tccacaggct ggaggagtca ttcctcgtgg
     3301 aggaggataa gaagcacgag cggcatccaa tcttcggcaa cattgtcgac gaggttgcct
     3361 accacgagaa gtaccctacg atctaccatc tgcggaagaa gctcgtggac tccacagata
     3421 aggcggacct ccgcctgatc tacctcgctc tggcccacat gattaagttc aggggccatt
     3481 tcctgatcga gggggatctc aacccggaca atagcgatgt tgacaagctg ttcatccagc
     3541 tcgtgcagac gtacaaccag ctcttcgagg agaaccccat taatgcgtca ggcgtcgacg
     3601 cgaaggctat cctgtccgct aggctctcga agtctcggcg cctcgagaac ctgatcgccc
     3661 agctgccggg cgagaagaag aacggcctgt tcgggaatct cattgcgctc agcctggggc
     3721 tcacgcccaa cttcaagtcg aatttcgatc tcgctgagga cgccaagctg cagctctcca
     3781 aggacacata cgacgatgac ctggataacc tcctggccca gatcggcgat cagtacgcgg
     3841 acctgttcct cgctgccaag aatctgtcgg acgccatcct cctgtctgat attctcaggg
     3901 tgaacaccga gattacgaag gctccgctct cagcctccat gatcaagcgc tacgacgagc
     3961 accatcagga tctgaccctc ctgaaggcgc tggtcaggca gcagctcccc gagaagtaca
     4021 aggagatctt cttcgatcag tcgaagaacg gctacgctgg gtacattgac ggcggggcct
     4081 ctcaggagga gttctacaag ttcatcaagc cgattctgga gaagatggac ggcacggagg
     4141 agctgctggt gaagctcaat cgcgaggacc tcctgaggaa gcagcggaca ttcgataacg
     4201 gcagcatccc acaccagatt catctcgggg agctgcacgc tatcctgagg aggcaggagg
     4261 acttctaccc tttcctcaag gataaccgcg agaagatcga gaagattctg actttcagga
     4321 tcccgtacta cgtcggccca ctcgctaggg gcaactcccg cttcgcttgg atgacccgca
     4381 agtcagagga gacgatcacg ccgtggaact tcgaggaggt ggtcgacaag ggcgctagcg
     4441 ctcagtcgtt catcgagagg atgacgaatt tcgacaagaa cctgccaaat gagaaggtgc
     4501 tccctaagca ctcgctcctg tacgagtact tcacagtcta caacgagctg actaaggtga
     4561 agtatgtgac cgagggcatg aggaagccgg ctttcctgtc tggggagcag aagaaggcca
     4621 tcgtggacct cctgttcaag accaaccgga aggtcacggt taagcagctc aaggaggact
     4681 acttcaagaa gattgagtgc ttcgattcgg tcgagatctc tggcgttgag gaccgcttca
     4741 acgcctccct ggggacctac cacgatctcc tgaagatcat taaggataag gacttcctgg
     4801 acaacgagga gaatgaggat atcctcgagg acattgtgct gacactcact ctgttcgagg
     4861 accgggagat gatcgaggag cgcctgaaga cttacgccca tctcttcgat gacaaggtca
     4921 tgaagcagct caagaggagg aggtacaccg gctgggggag gctgagcagg aagctcatca
     4981 acggcattcg ggacaagcag tccgggaaga cgatcctcga cttcctgaag agcgatggct
     5041 tcgcgaaccg caatttcatg cagctgattc acgatgacag cctcacattc aaggaggata
     5101 tccagaaggc tcaggtgagc ggccaggggg actcgctgca cgagcatatc gcgaacctcg
     5161 ctggctcgcc agctatcaag aaggggattc tgcagaccgt gaaggttgtg gacgagctgg
     5221 tgaaggtcat gggcaggcac aagcctgaga acatcgtcat tgagatggcc cgggagaatc
     5281 agaccacgca gaagggccag aagaactcac gcgagaggat gaagaggatc gaggagggca
     5341 ttaaggagct ggggtcccag atcctcaagg agcacccggt ggagaacacg cagctgcaga
     5401 atgagaagct ctacctgtac tacctccaga atggccgcga tatgtatgtg gaccaggagc
     5461 tggatattaa caggctcagc gattacgacg tcgatcatat cgttccacag tcattcctga
     5521 aggatgactc cattgacaac aaggtcctca ccaggtcgga caagaaccgg ggcaagtctg
     5581 ataatgttcc ttcagaggag gtcgttaaga agatgaagaa ctactggcgc cagctcctga
     5641 atgccaagct gatcacgcag cggaagttcg ataacctcac aaaggctgag aggggcgggc
     5701 tctctgagct ggacaaggcg ggcttcatca agaggcagct ggtcgagaca cggcagatca
     5761 ctaagcacgt tgcgcagatt ctcgactcac ggatgaacac taagtacgat gagaatgaca
     5821 agctgatccg cgaggtgaag gtcatcaccc tgaagtcaaa gctcgtctcc gacttcagga
     5881 aggatttcca gttctacaag gttcgggaga tcaacaatta ccaccatgcc catgacgcgt
     5941 acctgaacgc ggtggtcggc acagctctga tcaagaagta cccaaagctc gagagcgagt
     6001 tcgtgtacgg ggactacaag gtttacgatg tgaggaagat gatcgccaag tcggagcagg
     6061 agattggcaa ggctaccgcc aagtacttct tctactctaa cattatgaat ttcttcaaga
     6121 cagagatcac tctggccaat ggcgagatcc ggaagcgccc cctcatcgag acgaacggcg
     6181 agacggggga gatcgtgtgg gacaagggca gggatttcgc gaccgtcagg aaggttctct
     6241 ccatgccaca agtgaatatc gtcaagaaga cagaggtcca gactggcggg ttctctaagg
     6301 agtcaattct gcctaagcgg aacagcgaca agctcatcgc ccgcaagaag gactgggatc
     6361 cgaagaagta cggcgggttc gacagcccca ctgtggccta ctcggtcctg gttgtggcga
     6421 aggttgagaa gggcaagtcc aagaagctca agagcgtgaa ggagctgctg gggatcacga
     6481 ttatggagcg ctccagcttc gagaagaacc cgatcgattt cctggaggcg aagggctaca
     6541 aggaggtgaa gaaggacctg atcattaagc tccccaagta ctcactcttc gagctggaga
     6601 acggcaggaa gcggatgctg gcttccgctg gcgagctgca gaaggggaac gagctggctc
     6661 tgccgtccaa gtatgtgaac ttcctctacc tggcctccca ctacgagaag ctcaagggca
     6721 gccccgagga caacgagcag aagcagctgt tcgtcgagca gcacaagcat tacctcgacg
     6781 agatcattga gcagatttcc gagttctcca agcgcgtgat cctggccgac gcgaatctgg
     6841 ataaggtcct ctccgcgtac aacaagcacc gcgacaagcc aatcagggag caggctgaga
     6901 atatcattca tctcttcacc ctgacgaacc tcggcgcccc tgctgctttc aagtacttcg
     6961 acacaactat cgatcgcaag aggtacacaa gcactaagga ggtcctggac gcgaccctca
     7021 tccaccagtc gattaccggc ctctacgaga cgcgcatcga cctgtctcag ctcgggggcg
     7081 acaagcggcc agcggcgacg aagaaggcgg ggcaggcgaa gaagaagaag tgagctcaga
     7141 gctttcgttc gtatcatcgg tttcgacaac gttcgtcaag ttcaatgcat cagtttcatt
     7201 gcgcacacac cagaatccta ctgagtttga gtattatggc attgggaaaa ctgtttttct
     7261 tgtaccattt gttgtgcttg taatttactg tgttttttat tcggttttcg ctatcgaact
     7321 gtgaaatgga aatggatgga gaagagttaa tgaatgatat ggtccttttg ttcattctca
     7381 aattaatatt atttgttttt tctcttattt gttgtgtgtt gaatttgaaa ttataagaga
     7441 tatgcaaaca ttttgttttg agtaaaaatg tgtcaaatcg tggcctctaa tgaccgaagt
     7501 taatatgagg agtaaaacac ttgtagttgt accattatgc ttattcacta ggcaacaaat
     7561 atattttcag acctagaaaa gctgcaaatg ttactgaata caagtatgtc ctcttgtgtt
     7621 ttagacattt atgaactttc ctttatgtaa ttttccagaa tccttgtcag attctaatca
     7681 ttgctttata attatagtta tactcatgga tttgtagttg agtatgaaaa tattttttaa
     7741 tgcattttat gacttgccaa ttgattgaca acgaattcgt aatcatgtca tagctgtttc
     7801 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     7861 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     7921 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     7981 ggagaggcgg tttgcgtatt ggctagagca gcttgccaac atggtggagc acgacactct
     8041 cgtctactcc aagaatatca aagatacagt ctcagaagac caaagggcta ttgagacttt
     8101 tcaacaaagg gtaatatcgg gaaacctcct cggattccat tgcccagcta tctgtcactt
     8161 catcaaaagg acagtagaaa aggaaggtgg cacctacaaa tgccatcatt gcgataaagg
     8221 aaaggctatc gttcaagatg cctctgccga cagtggtccc aaagatggac ccccacccac
     8281 gaggagcatc gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag tggattgatg
     8341 tgaacatggt ggagcacgac actctcgtct actccaagaa tatcaaagat acagtctcag
     8401 aagaccaaag ggctattgag acttttcaac aaagggtaat atcgggaaac ctcctcggat
     8461 tccattgccc agctatctgt cacttcatca aaaggacagt agaaaaggaa ggtggcacct
     8521 acaaatgcca tcattgcgat aaaggaaagg ctatcgttca agatgcctct gccgacagtg
     8581 gtcccaaaga tggaccccca cccacgagga gcatcgtgga aaaagaagac gttccaacca
     8641 cgtcttcaaa gcaagtggat tgatgtgata tctccactga cgtaagggat gacgcacaat
     8701 cccactatcc ttcgcaagac ccttcctcta tataaggaag ttcatttcat ttggagagga
     8761 cacgctgaaa tcaccagtct ctctctacaa atctatctct ctcgagcttt cgcagatctg
     8821 tcgatcgacc atggggattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     8881 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     8941 gtttcggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     9001 cctgaatgaa ctccaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     9061 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     9121 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     9181 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     9241 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     9301 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     9361 gcgcatgccc gacggcgagg atctcgtcgt gacacatggc gatgcctgct tgccgaatat
     9421 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     9481 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     9541 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     9601 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcggatc gatcctctag
     9661 ctagagtcga tcgacaagct cgagtttctc cataataatg tgtgagtagt tcccagataa
     9721 gggaattagg gttcctatag ggtttcgctc atgtgttgag catataagaa acccttagta
     9781 tgtatttgta tttgtaaaat acttctatca ataaaatttc taattcctaa aaccaaaatc
     9841 cagtactaaa atccagatcc cccgaattaa ttcggcgtta attcagtaca ttaaaaacgt
     9901 ccgcaatgtg ttattaagtt gtctaagcgt caatttgttt acaccacaat atatcctgcc
     9961 accagccagc caacagctcc ccgaccggca gctcggcaca aaatcaccac tcgatacagg
    10021 cagcccatca gtccgggacg gcgtcagcgg gagagccgtt gtaaggcggc agactttgct
    10081 catgttaccg atgctattcg gaagaacggc aactaagctg ccgggtttga aacacggatg
    10141 atctcgcgga gggtagcatg ttgattgtaa cgatgacaga gcgttgctgc ctgtgatcac
    10201 cgcggctggc aaacagctat tatgggtatt atgggtctag tctctggccc gtgtctcaaa
    10261 atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg
    10321 cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc
    10381 cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg
    10441 tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt
    10501 ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa
    10561 actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg
    10621 atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat
    10681 atcctgattc aggtgaaaat attgttgatg cgctggcagt gtccctgcgc cggttgcatt
    10741 cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc
    10801 aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct
    10861 ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag
    10921 tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag
    10981 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat
    11041 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta
    11101 ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat
    11161 cagaattggt taattggttg taacattatt cagattggcg tctaactact aagcgagagt
    11221 agggaactgc caggcatcaa ataaaacgaa aggctcagtc ggaagactgg gcctttcgtt
    11281 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt
    11341 tgaacgttgt gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca
    11401 ggcatcaaac taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc
    11461 ttcctgttag ggccccgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
    11521 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
    11581 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
    11641 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
    11701 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
    11761 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
    11821 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
    11881 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
    11941 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
    12001 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
    12061 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
    12121 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
    12181 cgttatcccc tgattctgtg gataaccgta ttaccgctag atagctggtt gccctcgccg
    12241 ctgggctggc ggccgtctat ggccctgcaa acgcgccaga aacgccgtcg aagccgtgtg
    12301 cgagacaccc cggccgccgg cgttgtggat acctcgcgga aaacttggcc ctcactgaca
    12361 gatgaggggc ggacgttgac acttgagggg ccgactcacc cggcgcggcg ttgacagatg
    12421 aggggcaggc tcgatttcgg ccggcgacgt ggagctggcc agcctcgcaa atcggcgaaa
    12481 acgcctgatt ttacgcgagt ttcccacaga tgatgtggac aagcctgggg ataagtgccc
    12541 tgcggtattg acacttgagg ggcgcgacta ctgacagatg aggggcgcga tccttgacac
    12601 ttgaggggca gagtgctgac agatgagggg cgcacctatt gacatttgag gggctgtcca
    12661 caggcagaaa atccagcatt tgcaagggtt tccgcccgtt tttcggccac cgctaacctg
    12721 tcttttaacc tgcttttaaa ccaatattta taaaccttgt ttttaaccag ggctgcgccc
    12781 tgtgcgcgtg accgcgcacg ccgaaggggg gtgccccccc ttctcgaacc ctcccggtcg
    12841 agtcatggtc tagcatggat ctcggggacg tcacccataa tacccataat agctgtttgc
    12901 cagtactttg atcccgaggg gaaccctgtg gttggcatgc acatacaaat ggacgaacgg
    12961 ataaaccttt tcacgccctt ttaaatatcc gattattcta ataaacgctc ttttctctta
    13021 g
//