pG3B-U6SC

LOCUS       pG3B-U6SC               12707 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  pGreen3 CRISPR/Cas9 .
ACCESSION   .
VERSION     .
KEYWORDS    pG3B-U6SC
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12707)
  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 12707)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..12707
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_RNA        364..439
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             1415..1480
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1487..1507
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1532..5632
                     /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             5545..5553
                     /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             5633..5680
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(6337..6353)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6337..6353)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6350..6372)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6361..6377
                     /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(6385..6415)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6430..6451
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6642..7318
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     7255..7275
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             7363..7914
                     /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    7921..8095
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    8173..8197
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     complement(8575..8594)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             8586..9395
                     /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(8653..8672)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      9474..9560
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      9652..9679
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      9758..10346
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10247..10266
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    11258..11282
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     primer_bind     11470..11492
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     11484..11501
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     11485..11501
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        11957..12061
                     /gene="bla"
                     /label=AmpR promoter
     CDS             12062..215
                     /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(12280..12299)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
       61 gtgagcgtgg ctctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
      121 tcgtagttat ctacaccacg gggagtcagg caactatgga tgaacgaaat agacagatcg
      181 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
      241 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
      301 ttgataatcc catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcaggtc
      361 tcagttttag agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa
      421 agtggcaccg agtcggtgct tttttttgga tagaatttcc cagctttttt gcgtgtttca
      481 gctctcatga tccttgcatg cactagtatg gagtcaaaga ttcaaataga ggacctaaca
      541 gaactcgccg taaagactgg cgaacagttc atacagagtc tcttacgact caatgacaag
      601 aagaaaatct tcgtcaacat ggtggagcac gacacacttg tctactccaa aaatatcaaa
      661 gatacagtct cagaagacca aagggcaatt gagacttttc aacaaagggt aatatccgga
      721 aacctcctcg gattccattg cccagctatc tgtcacttta ttgtgaagat agtggaaaag
      781 gaaggtggct cctacaaatg ccatcattgc gataaaggaa aggccatcgt tgaagatgcc
      841 tctgccgaca gtggtcccaa agatggaccc ccacccacga ggagcatcgt ggaaaaagaa
      901 gacgttccaa ccacgtcttc aaagcaagtg gattgatgtg attggcagac atactgtccc
      961 acaaatgaag atggaatctg taaaagaaaa cgcgtgaaat aatgcgtctg acaaaggtta
     1021 ggtcggctgc ctttaatcaa taccaaagtg gtccctacca cgatggaaaa actgtgcagt
     1081 cggtttggct ttttctgacg aacaaataag attcgtggcc gacaggtggg ggtccaccat
     1141 gtgaaggcat cttcagactc caataatgga gcaatgacgt aagggcttac gaaataagta
     1201 agggtagttt gggaaatgtc cactcacccg tcagtctata aatacttagc ccctccctca
     1261 ttgttaaggg agcaaaatct cagagagata gtcctagaga gagaaagaga gcaagtagcc
     1321 tagaagtagt caaggcggcg aagtattcag gcacgtggcc aggaagaaga aaagccaaga
     1381 cgacgaaaac aggtaagagc taagcatcta gatggattac aaggaccacg acggggatta
     1441 caaggaccac gacattgatt acaaggatga tgatgacaag atggctccga agaagaagag
     1501 gaaggttggc atccacgggg tgccagctgc tgacaagaag tactcgatcg gcctcgatat
     1561 tgggactaac tctgttggct gggccgtgat caccgacgag tacaaggtgc cctcaaagaa
     1621 gttcaaggtc ctgggcaaca ccgatcggca ttccatcaag aagaatctca ttggcgctct
     1681 cctgttcgac agcggcgaga cggctgaggc tacgcggctc aagcgcaccg cccgcaggcg
     1741 gtacacgcgc aggaagaatc gcatctgcta cctgcaggag attttctcca acgagatggc
     1801 gaaggttgac gattctttct tccacaggct ggaggagtca ttcctcgtgg aggaggataa
     1861 gaagcacgag cggcatccaa tcttcggcaa cattgtcgac gaggttgcct accacgagaa
     1921 gtaccctacg atctaccatc tgcggaagaa gctcgtggac tccacagata aggcggacct
     1981 ccgcctgatc tacctcgctc tggcccacat gattaagttc aggggccatt tcctgatcga
     2041 gggggatctc aacccggaca atagcgatgt tgacaagctg ttcatccagc tcgtgcagac
     2101 gtacaaccag ctcttcgagg agaaccccat taatgcgtca ggcgtcgacg cgaaggctat
     2161 cctgtccgct aggctctcga agtctcggcg cctcgagaac ctgatcgccc agctgccggg
     2221 cgagaagaag aacggcctgt tcgggaatct cattgcgctc agcctggggc tcacgcccaa
     2281 cttcaagtcg aatttcgatc tcgctgagga cgccaagctg cagctctcca aggacacata
     2341 cgacgatgac ctggataacc tcctggccca gatcggcgat cagtacgcgg acctgttcct
     2401 cgctgccaag aatctgtcgg acgccatcct cctgtctgat attctcaggg tgaacaccga
     2461 gattacgaag gctccgctct cagcctccat gatcaagcgc tacgacgagc accatcagga
     2521 tctgaccctc ctgaaggcgc tggtcaggca gcagctcccc gagaagtaca aggagatctt
     2581 cttcgatcag tcgaagaacg gctacgctgg gtacattgac ggcggggcct ctcaggagga
     2641 gttctacaag ttcatcaagc cgattctgga gaagatggac ggcacggagg agctgctggt
     2701 gaagctcaat cgcgaggacc tcctgaggaa gcagcggaca ttcgataacg gcagcatccc
     2761 acaccagatt catctcgggg agctgcacgc tatcctgagg aggcaggagg acttctaccc
     2821 tttcctcaag gataaccgcg agaagatcga gaagattctg actttcagga tcccgtacta
     2881 cgtcggccca ctcgctaggg gcaactcccg cttcgcttgg atgacccgca agtcagagga
     2941 gacgatcacg ccgtggaact tcgaggaggt ggtcgacaag ggcgctagcg ctcagtcgtt
     3001 catcgagagg atgacgaatt tcgacaagaa cctgccaaat gagaaggtgc tccctaagca
     3061 ctcgctcctg tacgagtact tcacagtcta caacgagctg actaaggtga agtatgtgac
     3121 cgagggcatg aggaagccgg ctttcctgtc tggggagcag aagaaggcca tcgtggacct
     3181 cctgttcaag accaaccgga aggtcacggt taagcagctc aaggaggact acttcaagaa
     3241 gattgagtgc ttcgattcgg tcgagatctc tggcgttgag gaccgcttca acgcctccct
     3301 ggggacctac cacgatctcc tgaagatcat taaggataag gacttcctgg acaacgagga
     3361 gaatgaggat atcctcgagg acattgtgct gacactcact ctgttcgagg accgggagat
     3421 gatcgaggag cgcctgaaga cttacgccca tctcttcgat gacaaggtca tgaagcagct
     3481 caagaggagg aggtacaccg gctgggggag gctgagcagg aagctcatca acggcattcg
     3541 ggacaagcag tccgggaaga cgatcctcga cttcctgaag agcgatggct tcgcgaaccg
     3601 caatttcatg cagctgattc acgatgacag cctcacattc aaggaggata tccagaaggc
     3661 tcaggtgagc ggccaggggg actcgctgca cgagcatatc gcgaacctcg ctggctcgcc
     3721 agctatcaag aaggggattc tgcagaccgt gaaggttgtg gacgagctgg tgaaggtcat
     3781 gggcaggcac aagcctgaga acatcgtcat tgagatggcc cgggagaatc agaccacgca
     3841 gaagggccag aagaactcac gcgagaggat gaagaggatc gaggagggca ttaaggagct
     3901 ggggtcccag atcctcaagg agcacccggt ggagaacacg cagctgcaga atgagaagct
     3961 ctacctgtac tacctccaga atggccgcga tatgtatgtg gaccaggagc tggatattaa
     4021 caggctcagc gattacgacg tcgatcatat cgttccacag tcattcctga aggatgactc
     4081 cattgacaac aaggtcctca ccaggtcgga caagaaccgg ggcaagtctg ataatgttcc
     4141 ttcagaggag gtcgttaaga agatgaagaa ctactggcgc cagctcctga atgccaagct
     4201 gatcacgcag cggaagttcg ataacctcac aaaggctgag aggggcgggc tctctgagct
     4261 ggacaaggcg ggcttcatca agaggcagct ggtcgagaca cggcagatca ctaagcacgt
     4321 tgcgcagatt ctcgactcac ggatgaacac taagtacgat gagaatgaca agctgatccg
     4381 cgaggtgaag gtcatcaccc tgaagtcaaa gctcgtctcc gacttcagga aggatttcca
     4441 gttctacaag gttcgggaga tcaacaatta ccaccatgcc catgacgcgt acctgaacgc
     4501 ggtggtcggc acagctctga tcaagaagta cccaaagctc gagagcgagt tcgtgtacgg
     4561 ggactacaag gtttacgatg tgaggaagat gatcgccaag tcggagcagg agattggcaa
     4621 ggctaccgcc aagtacttct tctactctaa cattatgaat ttcttcaaga cagagatcac
     4681 tctggccaat ggcgagatcc ggaagcgccc cctcatcgag acgaacggcg agacggggga
     4741 gatcgtgtgg gacaagggca gggatttcgc gaccgtcagg aaggttctct ccatgccaca
     4801 agtgaatatc gtcaagaaga cagaggtcca gactggcggg ttctctaagg agtcaattct
     4861 gcctaagcgg aacagcgaca agctcatcgc ccgcaagaag gactgggatc cgaagaagta
     4921 cggcgggttc gacagcccca ctgtggccta ctcggtcctg gttgtggcga aggttgagaa
     4981 gggcaagtcc aagaagctca agagcgtgaa ggagctgctg gggatcacga ttatggagcg
     5041 ctccagcttc gagaagaacc cgatcgattt cctggaggcg aagggctaca aggaggtgaa
     5101 gaaggacctg atcattaagc tccccaagta ctcactcttc gagctggaga acggcaggaa
     5161 gcggatgctg gcttccgctg gcgagctgca gaaggggaac gagctggctc tgccgtccaa
     5221 gtatgtgaac ttcctctacc tggcctccca ctacgagaag ctcaagggca gccccgagga
     5281 caacgagcag aagcagctgt tcgtcgagca gcacaagcat tacctcgacg agatcattga
     5341 gcagatttcc gagttctcca agcgcgtgat cctggccgac gcgaatctgg ataaggtcct
     5401 ctccgcgtac aacaagcacc gcgacaagcc aatcagggag caggctgaga atatcattca
     5461 tctcttcacc ctgacgaacc tcggcgcccc tgctgctttc aagtacttcg acacaactat
     5521 cgatcgcaag aggtacacaa gcactaagga ggtcctggac gcgaccctca tccaccagtc
     5581 gattaccggc ctctacgaga cgcgcatcga cctgtctcag ctcgggggcg acaagcggcc
     5641 agcggcgacg aagaaggcgg ggcaggcgaa gaagaagaag tgagctcaga gctttcgttc
     5701 gtatcatcgg tttcgacaac gttcgtcaag ttcaatgcat cagtttcatt gcgcacacac
     5761 cagaatccta ctgagtttga gtattatggc attgggaaaa ctgtttttct tgtaccattt
     5821 gttgtgcttg taatttactg tgttttttat tcggttttcg ctatcgaact gtgaaatgga
     5881 aatggatgga gaagagttaa tgaatgatat ggtccttttg ttcattctca aattaatatt
     5941 atttgttttt tctcttattt gttgtgtgtt gaatttgaaa ttataagaga tatgcaaaca
     6001 ttttgttttg agtaaaaatg tgtcaaatcg tggcctctaa tgaccgaagt taatatgagg
     6061 agtaaaacac ttgtagttgt accattatgc ttattcacta ggcaacaaat atattttcag
     6121 acctagaaaa gctgcaaatg ttactgaata caagtatgtc ctcttgtgtt ttagacattt
     6181 atgaactttc ctttatgtaa ttttccagaa tccttgtcag attctaatca ttgctttata
     6241 attatagtta tactcatgga tttgtagttg agtatgaaaa tattttttaa tgcattttat
     6301 gacttgccaa ttgattgaca acgaattcgt aatcatgtca tagctgtttc ctgtgtgaaa
     6361 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     6421 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     6481 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6541 tttgcgtatt ggctagagca gcttgccaac atggtggagc acgacactct cgtctactcc
     6601 aagaatatca aagatacagt ctcagaagac caaagggcta ttgagacttt tcaacaaagg
     6661 gtaatatcgg gaaacctcct cggattccat tgcccagcta tctgtcactt catcaaaagg
     6721 acagtagaaa aggaaggtgg cacctacaaa tgccatcatt gcgataaagg aaaggctatc
     6781 gttcaagatg cctctgccga cagtggtccc aaagatggac ccccacccac gaggagcatc
     6841 gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag tggattgatg tgaacatggt
     6901 ggagcacgac actctcgtct actccaagaa tatcaaagat acagtctcag aagaccaaag
     6961 ggctattgag acttttcaac aaagggtaat atcgggaaac ctcctcggat tccattgccc
     7021 agctatctgt cacttcatca aaaggacagt agaaaaggaa ggtggcacct acaaatgcca
     7081 tcattgcgat aaaggaaagg ctatcgttca agatgcctct gccgacagtg gtcccaaaga
     7141 tggaccccca cccacgagga gcatcgtgga aaaagaagac gttccaacca cgtcttcaaa
     7201 gcaagtggat tgatgtgata tctccactga cgtaagggat gacgcacaat cccactatcc
     7261 ttcgcaagac ccttcctcta tataaggaag ttcatttcat ttggagagga cacgctgaaa
     7321 tcaccagtct ctctctacaa atctatctct ctcgagtcta ccatgagccc agaacgacgc
     7381 ccggccgaca tccgccgtgc caccgaggcg gacatgccgg cggtctgcac catcgtcaac
     7441 cactacatcg agacaagcac ggtcaacttc cgtaccgagc cgcaggaacc gcaggagtgg
     7501 acggacgacc tcgtccgtct gcgggagcgc tatccctggc tcgtcgccga ggtggacggc
     7561 gaggtcgccg gcatcgccta cgcgggcccc tggaaggcac gcaacgccta cgactggacg
     7621 gccgagtcga ccgtgtacgt ctccccccgc caccagcgga cgggactggg ctccacgctc
     7681 tacacccacc tgctgaagtc cctggaggca cagggcttca agagcgtggt cgctgtcatc
     7741 gggctgccca acgacccgag cgtgcgcatg cacgaggcgc tcggatatgc cccacgcggc
     7801 atgctgcggg cggccggctt caagcacggg aactggcatg acgtgggttt ctggcagctg
     7861 gacttcagcc tgccggtacc gccccgtccg gtcctgcccg tcaccgagat ttgactcgag
     7921 tttctccata ataatgtgtg agtagttccc agataaggga attagggttc ctatagggtt
     7981 tcgctcatgt gttgagcata taagaaaccc ttagtatgta tttgtatttg taaaatactt
     8041 ctatcaataa aatttctaat tcctaaaacc aaaatccagt actaaaatcc agatcccccg
     8101 aattaattcg gcgttaattc agtacattaa aaacgtccgc aatgtgttat taagttgtct
     8161 aagcgtcaat ttgtttacac cacaatatat cctgccacca gccagccaac agctccccga
     8221 ccggcagctc ggcacaaaat caccactcga tacaggcagc ccatcagtcc gggacggcgt
     8281 cagcgggaga gccgttgtaa ggcggcagac tttgctcatg ttaccgatgc tattcggaag
     8341 aacggcaact aagctgccgg gtttgaaaca cggatgatct cgcggagggt agcatgttga
     8401 ttgtaacgat gacagagcgt tgctgcctgt gatcaccgcg gctggcaaac agctattatg
     8461 ggtattatgg gtctagtctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag
     8521 ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg
     8581 gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg
     8641 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
     8701 atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg
     8761 ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc
     8821 ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga
     8881 tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg
     8941 ttgatgcgct ggcagtgtcc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt
     9001 ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg
     9061 ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
     9121 aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
     9181 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
     9241 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
     9301 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
     9361 tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac
     9421 attattcaga ttggcgtcta actactaagc gagagtaggg aactgccagg catcaaataa
     9481 aacgaaaggc tcagtcggaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
     9541 ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgtgaag caacggcccg
     9601 gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaactaag cagaaggcca
     9661 tcctgacgga tggccttttt gcgtttctac aaactcttcc tgttagggcc ccgttccact
     9721 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     9781 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     9841 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     9901 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     9961 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
    10021 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
    10081 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
    10141 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
    10201 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
    10261 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
    10321 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
    10381 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
    10441 accgtattac cgctagatag ctggttgccc tcgccgctgg gctggcggcc gtctatggcc
    10501 ctgcaaacgc gccagaaacg ccgtcgaagc cgtgtgcgag acaccccggc cgccggcgtt
    10561 gtggatacct cgcggaaaac ttggccctca ctgacagatg aggggcggac gttgacactt
    10621 gaggggccga ctcacccggc gcggcgttga cagatgaggg gcaggctcga tttcggccgg
    10681 cgacgtggag ctggccagcc tcgcaaatcg gcgaaaacgc ctgattttac gcgagtttcc
    10741 cacagatgat gtggacaagc ctggggataa gtgccctgcg gtattgacac ttgaggggcg
    10801 cgactactga cagatgaggg gcgcgatcct tgacacttga ggggcagagt gctgacagat
    10861 gaggggcgca cctattgaca tttgaggggc tgtccacagg cagaaaatcc agcatttgca
    10921 agggtttccg cccgtttttc ggccaccgct aacctgtctt ttaacctgct tttaaaccaa
    10981 tatttataaa ccttgttttt aaccagggct gcgccctgtg cgcgtgaccg cgcacgccga
    11041 aggggggtgc ccccccttct cgaaccctcc cggtcgagtc atggtctagc atggatctcg
    11101 gggacgtcac ccataatacc cataatagct gtttgccagt actttgatcc cgaggggaac
    11161 cctgtggttg gcatgcacat acaaatggac gaacggataa accttttcac gcccttttaa
    11221 atatccgatt attctaataa acgctctttt ctcttaggtt tacccgccaa tatatcctgt
    11281 caaacactga tagtttaaac tgaaggcggg aaacgacaat ctgatccaag ctcaagctgc
    11341 tctagcattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
    11401 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
    11461 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctt cgacttgcct
    11521 tccgcacaat acatcatttc ttcttagctt tttttcttct tcttcgttca tacagttttt
    11581 ttttgtttat cagcttacat tttcttgaac cgtagctttc gttttcttct ttttaacttt
    11641 ccattcggag tttttgtatc ttgtttcata gtttgtccca ggattagaat gattaggcat
    11701 cgaaccttca agaatttgat tgaataaaac atcttcattc ttaagatatg aagataatct
    11761 tcaaaaggcc cctgggaatc tgaaagaaga gaagcaggcc catttatatg ggaaagaaca
    11821 atagtatttc ttatataggc ccatttaagt tgaaaacaat cttcaaaagt cccacatcgc
    11881 ttagataaga aaacgaagct gagtttatat acagctagag tcgaagtagt gattgtgaga
    11941 ccttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
    12001 gtatccgctc atgggacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
    12061 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
    12121 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
    12181 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
    12241 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
    12301 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
    12361 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
    12421 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
    12481 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
    12541 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
    12601 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
    12661 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcag
//