pG3B-U6EC1

LOCUS       pG3B-U6EC1               13155 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  pGreen3 CRISPR/Cas9 .
ACCESSION   .
VERSION     .
KEYWORDS    pG3B-U6EC1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13155)
  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 13155)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..13155
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        473..577
                     /gene="bla"
                     /label=AmpR promoter
     CDS             578..1438
                     /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(796..815)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     misc_RNA        1587..1662
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             3086..3151
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3158..3178
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3203..7303
                     /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             7216..7224
                     /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             7304..7351
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(8008..8024)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8008..8024)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8021..8043)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8032..8048
                     /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(8056..8086)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8101..8122
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8313..8989
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     8926..8946
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             9034..9585
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     polyA_signal    9592..9766
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    9844..9868
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     complement(10246..10265)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             10257..11066
                     /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(10324..10343)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      11145..11231
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      11323..11350
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      11429..12017
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11918..11937
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    12929..12953
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     13141..8
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     13155..17
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgcc aagcttcgac ttgccttccg cacaatacat catttcttct
       61 tagctttttt tcttcttctt cgttcataca gttttttttt gtttatcagc ttacattttc
      121 ttgaaccgta gctttcgttt tcttcttttt aactttccat tcggagtttt tgtatcttgt
      181 ttcatagttt gtcccaggat tagaatgatt aggcatcgaa ccttcaagaa tttgattgaa
      241 taaaacatct tcattcttaa gatatgaaga taatcttcaa aaggcccctg ggaatctgaa
      301 agaagagaag caggcccatt tatatgggaa agaacaatag tatttcttat ataggcccat
      361 ttaagttgaa aacaatcttc aaaagtccca catcgcttag ataagaaaac gaagctgagt
      421 ttatatacag ctagagtcga agtagtgatt gtgagacctt cggggaaatg tgcgcggaac
      481 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatgg gacaataacc
      541 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
      601 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
      661 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
      721 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
      781 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
      841 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
      901 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
      961 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     1021 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     1081 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     1141 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     1201 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     1261 tattgctgat aaatctggag ccggtgagcg tggctctcgc ggtatcattg cagcactggg
     1321 gccagatggt aagccctccc gtatcgtagt tatctacacc acggggagtc aggcaactat
     1381 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     1441 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     1501 aaggatctag gtgaagatcc tttttgataa tcccatgacc aaaatccctt aacgtgagtt
     1561 ttcgttccac tgagcgtcag gtctcagttt tagagctaga aatagcaagt taaaataagg
     1621 ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gctttttttt ggatagaatt
     1681 tcccagcttt tttgcgtgtt tcagctctca tgatccttac tagtgaataa aagcatttgc
     1741 gtttggttta tcattgcgtt tatacaagga cagagatcca ctgagctgga atagcttaaa
     1801 accattatca gaacaaaata aaccattttt tgttaagaat cagagcatag taaacaacag
     1861 aaacaaccta agagaggtaa cttgtccaag aagatagcta attatatcta ttttataaaa
     1921 gttatcatag tttgtaagtc acaaaagatg caaataacag agaaactagg agacttgaga
     1981 atatacattc ttgtatattt gtattcgaga ttgtgaaaat ttgaccataa gtttaaattc
     2041 ttaaaaagat atatctgatc taggtgatgg ttatagactg taattttacc acatgtttaa
     2101 tgatggatag tgacacacat gacacatcga caacactata gcatcttatt tagattacaa
     2161 catgaaattt ttctgtaata catgtctttg tacataattt aaaagtaatt cctaagaaat
     2221 atatttatac aaggagttta aagaaaacat agcataaagt tcaatgagta gtaaaaacca
     2281 tatacagtat atagcataaa gttcaatgag tttattacaa aagcattggt tcactttctg
     2341 taacacgacg ttaaaccttc gtctccaata ggagcgctac tgattcaaca tgccaatata
     2401 tactaaatac gtttctacag tcaaatgctt taacgtttca tgattaagtg actatttacc
     2461 gtcaatcctt tcccattcct cccactaatc caacttttta attactctta aatcaccact
     2521 aagctagtaa cgcctatcat gaattagctc tactaaatct agcaaccttt caaatttgca
     2581 gtattgcagg tgtctctgtg tctttaaaat agttgcctta tgatttcttc ggtttcaaga
     2641 tgatcaaata gttatagatt tcatgctcac acatgctcat tagatgtgta catactttac
     2701 ttacccaaat ctattttctc gcaaagattt tgatggtaaa gctgatttgg ttctattgaa
     2761 ctaaatcaaa cgagtttcag actgagtgat tctaatccgg cccattagcc cctaaacaga
     2821 cccactaatt acgcagcttt taatagagta attacaccta gtttacccac taaaccacta
     2881 agcactaatt atctcacaat ctaatgagct tccctcgtaa ttacttgggc tttcactcta
     2941 ccatttattt gtaacagtca agtctctact gtctctatat aaactctcta aagttaacac
     3001 acaattctca tcacaaacaa atcaaccaaa gcaacttcta ctctttcttc tttcgacctt
     3061 atcaatctgt tgagaaatct agatggatta caaggaccac gacggggatt acaaggacca
     3121 cgacattgat tacaaggatg atgatgacaa gatggctccg aagaagaaga ggaaggttgg
     3181 catccacggg gtgccagctg ctgacaagaa gtactcgatc ggcctcgata ttgggactaa
     3241 ctctgttggc tgggccgtga tcaccgacga gtacaaggtg ccctcaaaga agttcaaggt
     3301 cctgggcaac accgatcggc attccatcaa gaagaatctc attggcgctc tcctgttcga
     3361 cagcggcgag acggctgagg ctacgcggct caagcgcacc gcccgcaggc ggtacacgcg
     3421 caggaagaat cgcatctgct acctgcagga gattttctcc aacgagatgg cgaaggttga
     3481 cgattctttc ttccacaggc tggaggagtc attcctcgtg gaggaggata agaagcacga
     3541 gcggcatcca atcttcggca acattgtcga cgaggttgcc taccacgaga agtaccctac
     3601 gatctaccat ctgcggaaga agctcgtgga ctccacagat aaggcggacc tccgcctgat
     3661 ctacctcgct ctggcccaca tgattaagtt caggggccat ttcctgatcg agggggatct
     3721 caacccggac aatagcgatg ttgacaagct gttcatccag ctcgtgcaga cgtacaacca
     3781 gctcttcgag gagaacccca ttaatgcgtc aggcgtcgac gcgaaggcta tcctgtccgc
     3841 taggctctcg aagtctcggc gcctcgagaa cctgatcgcc cagctgccgg gcgagaagaa
     3901 gaacggcctg ttcgggaatc tcattgcgct cagcctgggg ctcacgccca acttcaagtc
     3961 gaatttcgat ctcgctgagg acgccaagct gcagctctcc aaggacacat acgacgatga
     4021 cctggataac ctcctggccc agatcggcga tcagtacgcg gacctgttcc tcgctgccaa
     4081 gaatctgtcg gacgccatcc tcctgtctga tattctcagg gtgaacaccg agattacgaa
     4141 ggctccgctc tcagcctcca tgatcaagcg ctacgacgag caccatcagg atctgaccct
     4201 cctgaaggcg ctggtcaggc agcagctccc cgagaagtac aaggagatct tcttcgatca
     4261 gtcgaagaac ggctacgctg ggtacattga cggcggggcc tctcaggagg agttctacaa
     4321 gttcatcaag ccgattctgg agaagatgga cggcacggag gagctgctgg tgaagctcaa
     4381 tcgcgaggac ctcctgagga agcagcggac attcgataac ggcagcatcc cacaccagat
     4441 tcatctcggg gagctgcacg ctatcctgag gaggcaggag gacttctacc ctttcctcaa
     4501 ggataaccgc gagaagatcg agaagattct gactttcagg atcccgtact acgtcggccc
     4561 actcgctagg ggcaactccc gcttcgcttg gatgacccgc aagtcagagg agacgatcac
     4621 gccgtggaac ttcgaggagg tggtcgacaa gggcgctagc gctcagtcgt tcatcgagag
     4681 gatgacgaat ttcgacaaga acctgccaaa tgagaaggtg ctccctaagc actcgctcct
     4741 gtacgagtac ttcacagtct acaacgagct gactaaggtg aagtatgtga ccgagggcat
     4801 gaggaagccg gctttcctgt ctggggagca gaagaaggcc atcgtggacc tcctgttcaa
     4861 gaccaaccgg aaggtcacgg ttaagcagct caaggaggac tacttcaaga agattgagtg
     4921 cttcgattcg gtcgagatct ctggcgttga ggaccgcttc aacgcctccc tggggaccta
     4981 ccacgatctc ctgaagatca ttaaggataa ggacttcctg gacaacgagg agaatgagga
     5041 tatcctcgag gacattgtgc tgacactcac tctgttcgag gaccgggaga tgatcgagga
     5101 gcgcctgaag acttacgccc atctcttcga tgacaaggtc atgaagcagc tcaagaggag
     5161 gaggtacacc ggctggggga ggctgagcag gaagctcatc aacggcattc gggacaagca
     5221 gtccgggaag acgatcctcg acttcctgaa gagcgatggc ttcgcgaacc gcaatttcat
     5281 gcagctgatt cacgatgaca gcctcacatt caaggaggat atccagaagg ctcaggtgag
     5341 cggccagggg gactcgctgc acgagcatat cgcgaacctc gctggctcgc cagctatcaa
     5401 gaaggggatt ctgcagaccg tgaaggttgt ggacgagctg gtgaaggtca tgggcaggca
     5461 caagcctgag aacatcgtca ttgagatggc ccgggagaat cagaccacgc agaagggcca
     5521 gaagaactca cgcgagagga tgaagaggat cgaggagggc attaaggagc tggggtccca
     5581 gatcctcaag gagcacccgg tggagaacac gcagctgcag aatgagaagc tctacctgta
     5641 ctacctccag aatggccgcg atatgtatgt ggaccaggag ctggatatta acaggctcag
     5701 cgattacgac gtcgatcata tcgttccaca gtcattcctg aaggatgact ccattgacaa
     5761 caaggtcctc accaggtcgg acaagaaccg gggcaagtct gataatgttc cttcagagga
     5821 ggtcgttaag aagatgaaga actactggcg ccagctcctg aatgccaagc tgatcacgca
     5881 gcggaagttc gataacctca caaaggctga gaggggcggg ctctctgagc tggacaaggc
     5941 gggcttcatc aagaggcagc tggtcgagac acggcagatc actaagcacg ttgcgcagat
     6001 tctcgactca cggatgaaca ctaagtacga tgagaatgac aagctgatcc gcgaggtgaa
     6061 ggtcatcacc ctgaagtcaa agctcgtctc cgacttcagg aaggatttcc agttctacaa
     6121 ggttcgggag atcaacaatt accaccatgc ccatgacgcg tacctgaacg cggtggtcgg
     6181 cacagctctg atcaagaagt acccaaagct cgagagcgag ttcgtgtacg gggactacaa
     6241 ggtttacgat gtgaggaaga tgatcgccaa gtcggagcag gagattggca aggctaccgc
     6301 caagtacttc ttctactcta acattatgaa tttcttcaag acagagatca ctctggccaa
     6361 tggcgagatc cggaagcgcc ccctcatcga gacgaacggc gagacggggg agatcgtgtg
     6421 ggacaagggc agggatttcg cgaccgtcag gaaggttctc tccatgccac aagtgaatat
     6481 cgtcaagaag acagaggtcc agactggcgg gttctctaag gagtcaattc tgcctaagcg
     6541 gaacagcgac aagctcatcg cccgcaagaa ggactgggat ccgaagaagt acggcgggtt
     6601 cgacagcccc actgtggcct actcggtcct ggttgtggcg aaggttgaga agggcaagtc
     6661 caagaagctc aagagcgtga aggagctgct ggggatcacg attatggagc gctccagctt
     6721 cgagaagaac ccgatcgatt tcctggaggc gaagggctac aaggaggtga agaaggacct
     6781 gatcattaag ctccccaagt actcactctt cgagctggag aacggcagga agcggatgct
     6841 ggcttccgct ggcgagctgc agaaggggaa cgagctggct ctgccgtcca agtatgtgaa
     6901 cttcctctac ctggcctccc actacgagaa gctcaagggc agccccgagg acaacgagca
     6961 gaagcagctg ttcgtcgagc agcacaagca ttacctcgac gagatcattg agcagatttc
     7021 cgagttctcc aagcgcgtga tcctggccga cgcgaatctg gataaggtcc tctccgcgta
     7081 caacaagcac cgcgacaagc caatcaggga gcaggctgag aatatcattc atctcttcac
     7141 cctgacgaac ctcggcgccc ctgctgcttt caagtacttc gacacaacta tcgatcgcaa
     7201 gaggtacaca agcactaagg aggtcctgga cgcgaccctc atccaccagt cgattaccgg
     7261 cctctacgag acgcgcatcg acctgtctca gctcgggggc gacaagcggc cagcggcgac
     7321 gaagaaggcg gggcaggcga agaagaagaa gtgagctcag agctttcgtt cgtatcatcg
     7381 gtttcgacaa cgttcgtcaa gttcaatgca tcagtttcat tgcgcacaca ccagaatcct
     7441 actgagtttg agtattatgg cattgggaaa actgtttttc ttgtaccatt tgttgtgctt
     7501 gtaatttact gtgtttttta ttcggttttc gctatcgaac tgtgaaatgg aaatggatgg
     7561 agaagagtta atgaatgata tggtcctttt gttcattctc aaattaatat tatttgtttt
     7621 ttctcttatt tgttgtgtgt tgaatttgaa attataagag atatgcaaac attttgtttt
     7681 gagtaaaaat gtgtcaaatc gtggcctcta atgaccgaag ttaatatgag gagtaaaaca
     7741 cttgtagttg taccattatg cttattcact aggcaacaaa tatattttca gacctagaaa
     7801 agctgcaaat gttactgaat acaagtatgt cctcttgtgt tttagacatt tatgaacttt
     7861 cctttatgta attttccaga atccttgtca gattctaatc attgctttat aattatagtt
     7921 atactcatgg atttgtagtt gagtatgaaa atatttttta atgcatttta tgacttgcca
     7981 attgattgac aacgaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc
     8041 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     8101 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     8161 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     8221 tggctagagc agcttgccaa catggtggag cacgacactc tcgtctactc caagaatatc
     8281 aaagatacag tctcagaaga ccaaagggct attgagactt ttcaacaaag ggtaatatcg
     8341 ggaaacctcc tcggattcca ttgcccagct atctgtcact tcatcaaaag gacagtagaa
     8401 aaggaaggtg gcacctacaa atgccatcat tgcgataaag gaaaggctat cgttcaagat
     8461 gcctctgccg acagtggtcc caaagatgga cccccaccca cgaggagcat cgtggaaaaa
     8521 gaagacgttc caaccacgtc ttcaaagcaa gtggattgat gtgaacatgg tggagcacga
     8581 cactctcgtc tactccaaga atatcaaaga tacagtctca gaagaccaaa gggctattga
     8641 gacttttcaa caaagggtaa tatcgggaaa cctcctcgga ttccattgcc cagctatctg
     8701 tcacttcatc aaaaggacag tagaaaagga aggtggcacc tacaaatgcc atcattgcga
     8761 taaaggaaag gctatcgttc aagatgcctc tgccgacagt ggtcccaaag atggaccccc
     8821 acccacgagg agcatcgtgg aaaaagaaga cgttccaacc acgtcttcaa agcaagtgga
     8881 ttgatgtgat atctccactg acgtaaggga tgacgcacaa tcccactatc cttcgcaaga
     8941 cccttcctct atataaggaa gttcatttca tttggagagg acacgctgaa atcaccagtc
     9001 tctctctaca aatctatctc tctcgagtct accatgagcc cagaacgacg cccggccgac
     9061 atccgccgtg ccaccgaggc ggacatgccg gcggtctgca ccatcgtcaa ccactacatc
     9121 gagacaagca cggtcaactt ccgtaccgag ccgcaggaac cgcaggagtg gacggacgac
     9181 ctcgtccgtc tgcgggagcg ctatccctgg ctcgtcgccg aggtggacgg cgaggtcgcc
     9241 ggcatcgcct acgcgggccc ctggaaggca cgcaacgcct acgactggac ggccgagtcg
     9301 accgtgtacg tctccccccg ccaccagcgg acgggactgg gctccacgct ctacacccac
     9361 ctgctgaagt ccctggaggc acagggcttc aagagcgtgg tcgctgtcat cgggctgccc
     9421 aacgacccga gcgtgcgcat gcacgaggcg ctcggatatg ccccacgcgg catgctgcgg
     9481 gcggccggct tcaagcacgg gaactggcat gacgtgggtt tctggcagct ggacttcagc
     9541 ctgccggtac cgccccgtcc ggtcctgccc gtcaccgaga tttgactcga gtttctccat
     9601 aataatgtgt gagtagttcc cagataaggg aattagggtt cctatagggt ttcgctcatg
     9661 tgttgagcat ataagaaacc cttagtatgt atttgtattt gtaaaatact tctatcaata
     9721 aaatttctaa ttcctaaaac caaaatccag tactaaaatc cagatccccc gaattaattc
     9781 ggcgttaatt cagtacatta aaaacgtccg caatgtgtta ttaagttgtc taagcgtcaa
     9841 tttgtttaca ccacaatata tcctgccacc agccagccaa cagctccccg accggcagct
     9901 cggcacaaaa tcaccactcg atacaggcag cccatcagtc cgggacggcg tcagcgggag
     9961 agccgttgta aggcggcaga ctttgctcat gttaccgatg ctattcggaa gaacggcaac
    10021 taagctgccg ggtttgaaac acggatgatc tcgcggaggg tagcatgttg attgtaacga
    10081 tgacagagcg ttgctgcctg tgatcaccgc ggctggcaaa cagctattat gggtattatg
    10141 ggtctagtct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata
    10201 tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga
    10261 gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat
    10321 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt
    10381 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg
    10441 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca
    10501 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atccccggaa
    10561 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc
    10621 tggcagtgtc cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg
    10681 atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga
    10741 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata
    10801 aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
    10861 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
    10921 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
    10981 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc
    11041 atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa cattattcag
    11101 attggcgtct aactactaag cgagagtagg gaactgccag gcatcaaata aaacgaaagg
    11161 ctcagtcgga agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
    11221 gtaggacaaa tccgccggga gcggatttga acgttgtgaa gcaacggccc ggagggtggc
    11281 gggcaggacg cccgccataa actgccaggc atcaaactaa gcagaaggcc atcctgacgg
    11341 atggcctttt tgcgtttcta caaactcttc ctgttagggc cccgttccac tgagcgtcag
    11401 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
    11461 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
    11521 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
    11581 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
    11641 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
    11701 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
    11761 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
    11821 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
    11881 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
    11941 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
    12001 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
    12061 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
    12121 ccgctagata gctggttgcc ctcgccgctg ggctggcggc cgtctatggc cctgcaaacg
    12181 cgccagaaac gccgtcgaag ccgtgtgcga gacaccccgg ccgccggcgt tgtggatacc
    12241 tcgcggaaaa cttggccctc actgacagat gaggggcgga cgttgacact tgaggggccg
    12301 actcacccgg cgcggcgttg acagatgagg ggcaggctcg atttcggccg gcgacgtgga
    12361 gctggccagc ctcgcaaatc ggcgaaaacg cctgatttta cgcgagtttc ccacagatga
    12421 tgtggacaag cctggggata agtgccctgc ggtattgaca cttgaggggc gcgactactg
    12481 acagatgagg ggcgcgatcc ttgacacttg aggggcagag tgctgacaga tgaggggcgc
    12541 acctattgac atttgagggg ctgtccacag gcagaaaatc cagcatttgc aagggtttcc
    12601 gcccgttttt cggccaccgc taacctgtct tttaacctgc ttttaaacca atatttataa
    12661 accttgtttt taaccagggc tgcgccctgt gcgcgtgacc gcgcacgccg aaggggggtg
    12721 cccccccttc tcgaaccctc ccggtcgagt catggtctag catggatctc ggggacgtca
    12781 cccataatac ccataatagc tgtttgccag tactttgatc ccgaggggaa ccctgtggtt
    12841 ggcatgcaca tacaaatgga cgaacggata aaccttttca cgccctttta aatatccgat
    12901 tattctaata aacgctcttt tctcttaggt ttacccgcca atatatcctg tcaaacactg
    12961 atagtttaaa ctgaaggcgg gaaacgacaa tctgatccaa gctcaagctg ctctagcatt
    13021 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
    13081 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
    13141 cccagtcacg acgtt
//