pG3K-U6EC1

LOCUS       pG3K-U6EC1               13469 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  pGreen3 CRISPR/Cas9 .
ACCESSION   .
VERSION     .
KEYWORDS    pG3K-U6EC1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13469)
  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 13469)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..13469
                     /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             9052..9849
                     /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(9109..9128)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     9719..9738
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    9906..10080
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    10158..10182
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     complement(10560..10579)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             10571..11380
                     /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(10638..10657)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      11459..11545
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      11637..11664
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      11743..12331
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12232..12251
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    13243..13267
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     13455..8
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     13469..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 tctcgagctt tcgcagatct gtcgatcgac catggggatt
     9061 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     9121 gactgggcac aacagacaat cggctgctct gatgccgccg tgtttcggct gtcagcgcag
     9181 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actccaggac
     9241 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     9301 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     9361 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     9421 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     9481 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     9541 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     9601 gatctcgtcg tgacacatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     9661 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     9721 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     9781 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     9841 ttcttctgag cgggactctg gggttcggat cgatcctcta gctagagtcg atcgacaagc
     9901 tcgagtttct ccataataat gtgtgagtag ttcccagata agggaattag ggttcctata
     9961 gggtttcgct catgtgttga gcatataaga aacccttagt atgtatttgt atttgtaaaa
    10021 tacttctatc aataaaattt ctaattccta aaaccaaaat ccagtactaa aatccagatc
    10081 ccccgaatta attcggcgtt aattcagtac attaaaaacg tccgcaatgt gttattaagt
    10141 tgtctaagcg tcaatttgtt tacaccacaa tatatcctgc caccagccag ccaacagctc
    10201 cccgaccggc agctcggcac aaaatcacca ctcgatacag gcagcccatc agtccgggac
    10261 ggcgtcagcg ggagagccgt tgtaaggcgg cagactttgc tcatgttacc gatgctattc
    10321 ggaagaacgg caactaagct gccgggtttg aaacacggat gatctcgcgg agggtagcat
    10381 gttgattgta acgatgacag agcgttgctg cctgtgatca ccgcggctgg caaacagcta
    10441 ttatgggtat tatgggtcta gtctctggcc cgtgtctcaa aatctctgat gttacattgc
    10501 acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac
    10561 aaggggtgtt atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat
    10621 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
    10681 aatctatcgc ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
    10741 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
    10801 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
    10861 tgcgatcccc ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
    10921 tattgttgat gcgctggcag tgtccctgcg ccggttgcat tcgattcctg tttgtaattg
    10981 tccttttaac agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
    11041 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
    11101 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
    11161 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
    11221 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
    11281 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
    11341 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
    11401 gtaacattat tcagattggc gtctaactac taagcgagag tagggaactg ccaggcatca
    11461 aataaaacga aaggctcagt cggaagactg ggcctttcgt tttatctgtt gtttgtcggt
    11521 gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg tgaagcaacg
    11581 gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ctaagcagaa
    11641 ggccatcctg acggatggcc tttttgcgtt tctacaaact cttcctgtta gggccccgtt
    11701 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
    11761 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
    11821 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
    11881 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
    11941 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
    12001 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
    12061 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
    12121 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
    12181 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
    12241 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
    12301 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
    12361 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
    12421 ggataaccgt attaccgcta gatagctggt tgccctcgcc gctgggctgg cggccgtcta
    12481 tggccctgca aacgcgccag aaacgccgtc gaagccgtgt gcgagacacc ccggccgccg
    12541 gcgttgtgga tacctcgcgg aaaacttggc cctcactgac agatgagggg cggacgttga
    12601 cacttgaggg gccgactcac ccggcgcggc gttgacagat gaggggcagg ctcgatttcg
    12661 gccggcgacg tggagctggc cagcctcgca aatcggcgaa aacgcctgat tttacgcgag
    12721 tttcccacag atgatgtgga caagcctggg gataagtgcc ctgcggtatt gacacttgag
    12781 gggcgcgact actgacagat gaggggcgcg atccttgaca cttgaggggc agagtgctga
    12841 cagatgaggg gcgcacctat tgacatttga ggggctgtcc acaggcagaa aatccagcat
    12901 ttgcaagggt ttccgcccgt ttttcggcca ccgctaacct gtcttttaac ctgcttttaa
    12961 accaatattt ataaaccttg tttttaacca gggctgcgcc ctgtgcgcgt gaccgcgcac
    13021 gccgaagggg ggtgcccccc cttctcgaac cctcccggtc gagtcatggt ctagcatgga
    13081 tctcggggac gtcacccata atacccataa tagctgtttg ccagtacttt gatcccgagg
    13141 ggaaccctgt ggttggcatg cacatacaaa tggacgaacg gataaacctt ttcacgccct
    13201 tttaaatatc cgattattct aataaacgct cttttctctt aggtttaccc gccaatatat
    13261 cctgtcaaac actgatagtt taaactgaag gcgggaaacg acaatctgat ccaagctcaa
    13321 gctgctctag cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
    13381 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
    13441 aacgccaggg ttttcccagt cacgacgtt
//