LOCUS       Cas9_2-4-6_FINAL       16132 bp    DNA     circular SYN 18-AUG-2025
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16132)
  AUTHORS   NC State
  TITLE     Direct Submission
  JOURNAL   Exported Aug 18, 2025 from SnapGene 8.0.2
            https://www.snapgene.com
COMMENT     Sequence Label: Cas9 tra246
FEATURES             Location/Qualifiers
     source          1..16132
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    201..351
                     /label=Lc U6b terminator
                     /note="color: #a6acb3"
     misc_RNA        complement(352..427)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
                     /note="color: #00ccff; direction: LEFT"
     misc_RNA        complement(526..601)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
                     /note="color: #00ccff; direction: LEFT"
     misc_RNA        complement(700..775)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
                     /note="color: #00ccff; direction: LEFT"
     misc_feature    796..866
                     /label=Dm tRNA Gly
                     /note="color: #33cccc"
     misc_feature    complement(893..1039)
                     /label=LcU6b promoter
                     /note="color: #a6acb3; direction: LEFT"
     misc_feature    893..1039
                     /label=Lc U6b promoter
                     /note="color: #a6acb3"
     misc_feature    1062..1569
                     /label=Tub 3' UTR
                     /note="color: #ffcc00"
     CDS             complement(1568..2245)
                     /codon_start=1
                     /label=RFPex
                     /note="color: #ff0000"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTFIYHVKFIGVNFPSDGLVMQKKTLGWEPSTERLYPRDGVLKGEIHK
                     ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     misc_feature    complement(2246..4460)
                     /label=Lc Hsp83
                     /note="color: #a6acb3; direction: LEFT"
     protein_bind    4542..4641
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
                     /note="color: #31849b"
     misc_feature    4572..4613
                     /label=attP
                     /note="color: #ffff00"
     misc_feature    5500..6258
                     /label=3' pBac
                     /note="color: #ff99cc"
     primer_bind     complement(6311..6327)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     promoter        6801..6905
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             join(6906..6974,6975..7766)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 6906 .. 6974 / #ccffcc / signal sequence
                        2: 6975 .. 7766 / #ccffcc
                     Cleavage site after base 6974"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      7937..8525
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     protein_bind    8813..8834
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     promoter        join(8849..8854,8855..8872,8873..8879)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This forward directional feature has 3 segments:
                        1: 8849 .. 8854 / #ffffff / -35
                        2: 8855 .. 8872 / #ffffff
                        3: 8873 .. 8879 / #ffffff / -10"
     protein_bind    8887..8903
                     /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)."
                     /note="color: #31849b"
     primer_bind     8911..8927
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     misc_feature    8983..9641
                     /label=5' pBac
                     /note="color: #ff99cc"
     protein_bind    9975..10074
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
                     /note="color: #31849b"
     misc_feature    10003..10044
                     /label=attP
                     /note="color: #ffff00"
     protein_bind    10209..10227
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    10230..10248
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    10251..10269
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    10272..10290
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    10293..10311
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    10314..10332
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    10335..10353
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    10356..10374
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    join(10484..10502,10503..10525,10526..10544,10545..10567,
                     10568..10586,10587..10609,10610..10628,10629..10651,
                     10652..10670,10671..10693,10694..10712,10713..10735,
                     10736..10754)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
                     /note="This feature has 13 segments:
                         1: 10484 .. 10502 / #31849b / tetO
                         2: 10503 .. 10525 / #31849b
                         3: 10526 .. 10544 / #31849b / tetO
                         4: 10545 .. 10567 / #31849b
                         5: 10568 .. 10586 / #31849b / tetO
                         6: 10587 .. 10609 / #31849b
                         7: 10610 .. 10628 / #31849b / tetO
                         8: 10629 .. 10651 / #31849b
                         9: 10652 .. 10670 / #31849b / tetO
                        10: 10671 .. 10693 / #31849b
                        11: 10694 .. 10712 / #31849b / tetO
                        12: 10713 .. 10735 / #31849b
                        13: 10736 .. 10754 / #31849b / tetO"
     protein_bind    10786..10804
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    10828..10846
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    10870..10888
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     misc_feature    10912..11200
                     /label=Lc Hsp70 promoter
                     /note="color: #a6acb3; direction: RIGHT"
     CDS             11214..11234
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /note="color: #993366"
                     /translation="PKKKRKV"
     CDS             11259..15359
                     /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"
                     /note="color: #993366"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYEVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             15360..15407
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /note="color: #993366"
                     /translation="KRPAATKKAGQAKKKK"
     CDS             15360..15407
                     /codon_start=1
                     /label=NLS
                     /note="color: #cc99ff"
                     /translation="KRPAATKKAGQAKKKK"
ORIGIN
        1 tactgtccgt ttttatttgt gtaataaaat gaacggaacg ctgaattctg gaacattaga
       61 aattctttgt tatagaatct ataaatttat gtcaaagttt tgtcgtttct tatgcgatga
      121 gagatcttaa aaggcaagta aattgagctg tcctattcaa acatttttct attaaattgt
      181 tggtaccttc cgcgtcgacg attcaatatc gagcaaaaat acgaacgcat atgattccat
      241 gaaacaaatg tatttcttta gacgtttcta aaaccaattc cgacaattat ttgattctat
      301 gacaactcca aatagttttg tttattaaaa aataaaattt caacaaaaaa agcaccgact
      361 cggtgccact ttttcaagtt gataacggac tagccttatt ttaacttgct atttctagct
      421 ctaaaacaac ctttatttga tcgcgattgc accagccggg aatcgaaccc gggtctgtac
      481 cgtggcaggg tactattcta ccactagacc actggtgctt tgttagcacc gactcggtgc
      541 cactttttca agttgataac ggactagcct tattttaact tgctatttct agctctaaaa
      601 cttacaacag gattggcagc atgcaccagc cgggaatcga acccgggtct gtaccgtggc
      661 agggtactat tctaccacta gaccactggt gctttgttag caccgactcg gtgccacttt
      721 ttcaagttga taacggacta gccttatttt aacttgctat ttctagctct aaaacgttta
      781 cagtcgattt taagttgcat cggccgggaa tcgaacccgg gccgcccgcg tggcaggcga
      841 gcattctacc actgaaccac cgatgcttga tgtctacaca cactcaaagc ccaaagatca
      901 tcactttaat aaactattta accgttcttt ttgagaaaaa agaatatggg aattataaac
      961 aacaatacta tatgttgaaa acataaaaat atcatttaca gattagggtt tgttattatc
     1021 acattattac tacctgagtc gtcgacgccg gatccttcaa gttcctcttg tacccatcaa
     1081 aatgaggcca aactagtgct gtttttaatg cactcactat gcatttttat agcttcctct
     1141 acacctcggc aagttcatta aacttttgcc aagcatttta acaattacta tttttagttg
     1201 ttttccctct ttttcccccc attaagtaat aaaacattaa aaaattttct tttggaatat
     1261 tttcattcaa tgcagcttaa gcctttcatt ttattaggtt cttgtttttt tcgtgttttt
     1321 aaagtgcact tctttaaaat ttgttctttt cattcatgtt ttttcaaatt tgaaaataaa
     1381 aaaaatttac aagtgttttg ttttggattc gttatcattt ttaaaataac agtttgttta
     1441 aaaaaaagta tattgaatgt ttagttgagt agtttcagct aataggtcga gaattttttc
     1501 aatacacact tcttctgtct tttttttggt tttgatggtt gtcgatgatg ggtttggctg
     1561 caagctttta ttgaaacaag tggtgacggg cttcggcacg ttcgtattgt tctactacgg
     1621 tgtaatcttc attatgactg gtaatatcca attttgaatc gacgtagtag taacctggca
     1681 acttaacggg ctttttggcc atgtagatgg acttaaattc aaccaagtag tgaccaccac
     1741 ccttcaattt caaagcctta tgaatttcac ctttcaagac accatcacgg ggatacaaac
     1801 gttcagtaga aggttcccaa cccaaggttt tcttttgcat aacaagacca tctgatggaa
     1861 agtttacacc gatgaatttg acgtggtaaa taaaagtacc atcttgcaaa gaggaatctt
     1921 gagtaacggt gacaacacca ccatcttcga aattcattac acgttcccac ttaaaacctt
     1981 cggggaagga caacttttta taatcgggaa tatcagcggg atgcttaacg tataccttag
     2041 aaccgtattg gaattgtggg gacaaaatat cccaggcgaa tggcaaggga ccacccttgg
     2101 taacttgcaa tttagcagtt tgagtacctt cgtagggttt accttcacct tcaccttcga
     2161 tttcaaattc gtgaccattg acagaacctt ccatgtgtac cttaaaacgc ataaaaggct
     2221 tgataacatt ttcagtagaa tccatggatc ctttaagctg agcaatttca gcttggaaag
     2281 caaaagtttc aacttcttca ggcatctgtg tttaataaaa aaaaaataga aaaatacaaa
     2341 ttattagtaa aataatcaat caaaatatga agtataaata attttaatga cgcacgtttt
     2401 tcgttaagtg tagggagtgg gattggtact gaaaattttt aataaaaatc tagaaatctc
     2461 tttaagttaa gaattactag ttttttaggg gtggattggg tatgcttttt aacgaatgaa
     2521 aacatgaaat atcgattata atttcgaaaa aattttgaaa aattttcttt aaaaaatcgc
     2581 atgctagaaa tttcttttta ttagcatggt gaaattttct ttatatacta gatttatttt
     2641 ctaaaaatgt catggccaaa gaaaatttaa tctagaacaa tttacaatta cttagtaacg
     2701 ttattcttaa accatattga aggtttttct ttctggcaaa ttacttgagt ttttacatca
     2761 cacttttttc aaaagaacaa aaacgttttg ttactacatt ttattttttt taataaaaac
     2821 ttaccttgat gtgtttgttt tatattaatt tcacaaaaat aaatactttt taaattattt
     2881 ttctttttgc aattgaaatt aaacaactac acactttttt aatgttcttt tttcagactt
     2941 ttctttatta agaaaattaa aacactttta actttctccc gtttggaatt cttggtgttt
     3001 tttgcacaat ttttctttca agactgatgc cggttagcag ccatgcactg aatttatagg
     3061 taaaacaaaa ataaacgaaa atactagaat gttctggaac aaactacccg tacttccgta
     3121 ggctctaaac aacatcatgt ggttgtttat ttggtgataa caaaagaaat tggttacatt
     3181 ttagatatat aaaaaaatta atttattaaa attgtagaaa accaatatta attataggtt
     3241 tttattgtta ttttaattta ttagttgcaa attatcaaaa aaatgtgcaa aacattcaac
     3301 atgattaatg cccacacagc gttgtatata tgtagaagca gtggctttta aactgtaaaa
     3361 ttgcatcaat gtcaaaattt ctacaatgtt ctctctgttt ttaataacat gcggaatgaa
     3421 tcaactataa tttactttat ttatttatgc aaaaatatat aaaagttatt aattaaatga
     3481 aaaaggaatt aatcttaggc attgctgagt aatgtcatga cataaactaa atatttttct
     3541 agtcaaacaa tgaacttatt tatttcaaaa acgttttatt ctttaaatta tagttaaaaa
     3601 gtcttattaa ttgacaaatc tatgtgtaat taagtgtttt ttgaactaag ttacaagctt
     3661 aatatgtatt tgattaattg aaatatgtat taactattaa tttttaaatt aaataaattt
     3721 aatattaatt atatattaaa ctttgtgctt tatatggaaa tagtatgtat gatttaatca
     3781 cattcgattt acatattagt catcataaat tataaatgaa agtaatttaa ttgttttatt
     3841 aaagaaaata ataacattat taaaacaata atattgctat caaaatttat aaaaatgcat
     3901 aaaatgaaaa aatattgatt ttctctcatc atgggggcat taataatttt ttcgccagtg
     3961 cagctgatgt gcagtaaaac atcagagttg ccggtcttga ataaaaacaa tgaaaagtgg
     4021 aataatttaa aaatgtgaca atcatatact tttaacatat tttataaaat ttgactgtaa
     4081 aaatttaaaa attaaaattt gcaaacatta tctcatacat attataacaa ctcttttgtt
     4141 tttatataaa tttaataaat gaaatcaaaa attctctcac aaatttgtaa tttccaaaag
     4201 tgtggccttt tattttcccc tttttaggta aaaatttcaa atatctacaa ccctgtaaat
     4261 gacagctgtt tgcgctcttt tatgtatttt ttttgtttat catgtcggtt ttttagttcg
     4321 cggtgcaatt tactcgaaat ttcttaaagt ttgttgaatt tttggcggta taattataat
     4381 attttgtttt agtgttcttt ttatcaaatt ataatgttta agtgaattat tttctgataa
     4441 tggtttcaac acgtgggccc atctgcagaa ttcggcttgt actagtcgcg ctcgcgcgac
     4501 tgacggtcgt aagcacccgc gtacgtgtcc accccggtca caaccccttg tgtcatgtcg
     4561 gcgaccctac gcccccaact gagagaactc aaaggttacc ccagttgggg cactactccc
     4621 gaaaaccgct tctgacctgg gaaaacgtga agccccgggg catccgctga gggttgccgc
     4681 cggggcttcg gtgtgtccgt cagtgagctc ctcaggtaat aataacaact acaaccaaaa
     4741 aacaaaagca acaacaacga caacttctac agggcaagta acaaaacata gtatttgtca
     4801 acgacacact atatgaatga tgatgatgat gcaacaacag caacaacagc aacaacagct
     4861 cgagtatcaa caacacaaca aaaaaaactt ttttaaaaag acacagctgc tggcaacaaa
     4921 ataccacaaa agatataaac taacgcagtt gccacacaca cacatacata cataaattaa
     4981 aatattatca aacacacaca tatacatata tcacaacata cctatataca aaaccacatt
     5041 tactttgata aaaggcgatt caacaaaaaa gtagtcgtca tcctccttca tttatccttg
     5101 catttcacgt gtatctgtgt gtgttataaa acctgctggc caaaaggcag gtaggtagtt
     5161 gacaacacaa atattttact tctttttcat catcagcatc aacaacagca ggagcaacat
     5221 attgtagaca cacactctct gtttcttcac ttcacacaca tacaagtaaa ttatgacagt
     5281 gaaaaagacg ggtatttcca agcttcagtt tcgtttactt taggtcatta agatgactgt
     5341 cctgatagct tgatatcaat ggaatcaacg ggattgataa atggaattac tgtgattgat
     5401 aaatagcttc gaaagacctc gttacttcag ggtacttaac aaaatttact gttggccgcg
     5461 aaccgcgcca tggcgccggc cgggagccgg cgcatgcgaa ggtcgcctat ggcattattg
     5521 tacggaatga taaacattgc ctgcataaat tcttttatta tatacagcca taatgtcagt
     5581 agcaacggag aaaaggttca aagtcgcaaa aaatttatga gaaaccttta catgagcctg
     5641 acgtcatcgt ttatgcgtaa gcgtttagaa gctcctactt tgaagagata tttgcgcgat
     5701 aatatctcta atattttgcc aaatgaagtg cctggtacat cagatgacag tactgaagag
     5761 ccagtaatga aaaaacgtac ttactgtact tactgcccct ctaaaataag gcgaaaggca
     5821 aatgcatcgt gcaaaaaatg caaaaaagtt atttgtcgag agcataatat tgatatgtgc
     5881 caaagttgtt tctgactgac taataagtat aatttgtttc tattatgtat aagttaagct
     5941 aattacttat tttataatac aacatgactg tttttaaagt acaaaataag tttatttttg
     6001 taaaagagag aatgtttaaa agttttgtta ctttatagaa gaaattttga gtttttgttt
     6061 ttttttaata aataaataaa cataaataaa ttgtttgttg aatttattat tagtatgtaa
     6121 gtgtaaatat aataaaactt aatatctatt caaattaata aataaacctc gatatacaga
     6181 ccgataaaac acatgcgtca attttacgca tgattatctt taacgtacgt cacaatatga
     6241 ttatctttct agggttaagg gcccgggatc cgatatctag atgcattcgc gaggtaccga
     6301 gctcgaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     6361 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     6421 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt
     6481 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa
     6541 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     6601 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     6661 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     6721 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     6781 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     6841 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     6901 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     6961 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     7021 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     7081 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     7141 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     7201 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     7261 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     7321 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     7381 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     7441 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     7501 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     7561 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     7621 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     7681 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     7741 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     7801 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     7861 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     7921 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     7981 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     8041 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     8101 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     8161 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     8221 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     8281 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     8341 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     8401 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     8461 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     8521 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     8581 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     8641 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     8701 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     8761 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     8821 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     8881 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     8941 agcttgcatg caggcctctg cagtcgacgg gcccaagctt cctttaaccc tagaaagata
     9001 gtctgcgtaa aattgacgca tgcattcttg aaatattgct ctctctttct aaatagcgcg
     9061 aatccgtcgc tgtgcattta ggacatctca gtcgccgctt ggagctcccg tgaggcgtgc
     9121 ttgtcaatgc ggtaagtgtc actgattttg aactataacg accgcgtgag tcaaaatgac
     9181 gcatgattat cttttacgtg acttttaaga tttaactcat acgataatta tattgttatt
     9241 tcatgttcta cttacgtgat aacttattat atatatattt tcttgttata gatatcgtga
     9301 ctaatatata ataaaatggg tagttcttta gacgatgagc atatcctctc tgctcttctg
     9361 caaagcgatg acgagcttgt tggtgaggat tctgacagtg aaatatcaga tcacgtaagt
     9421 gaagatgacg tccagagcga tacagaagaa gcgtttatag atgaggtaca tgaagtgcag
     9481 ccaacgtcaa gcggtagtga aatattagac gaacaaaatg ttattgaaca accaggttct
     9541 tcattggctt ctaacagaat cttgaccttg cgacagagga ctattagagg taagaataaa
     9601 cattgttggt caacttcaaa gtccacgagg cgtagcggag tggtaccagg ccctcgagtc
     9661 tgggcccatg catgggagct ctcccaagat cttatggtcg acctgcaggc aacattcatg
     9721 tgtgggtgat tacactcaca caaaaacaca cacattcaca caaacacaca cacttataaa
     9781 gatgagaaca tttaacaaaa acaattataa aactgatggc aatggagcaa tgatgtgatg
     9841 aagctgatga aagttttgaa tattcattca agagttaaaa acatgaacaa acatgcacaa
     9901 aattagtcga gactgacgga cacaccgaag ccccggcggc aaccctcagc ggatgccccg
     9961 gggcttcacg ttttcccagg tcagaagcgg ttttcgggag tagtgcccca actggggtaa
    10021 cctttgagtt ctctcagttg ggggcgtagg gtcgccgaca tgacacaagg ggttgtgacc
    10081 ggggtggaca cgtacgcggg tgcttacgac cgtcagtcgc gcgagcgcga ctagtacaag
    10141 ccgaattctg cagatggcgc gccaacctat gcttatccgt ttttatttgt gtaatggccg
    10201 cagaattctc tctatcactg atagggaggt ctctatcact gatagggagt tctctatcac
    10261 tgatagggat gtctctatca ctgataggga cgtctctatc actgataggg aactctctat
    10321 cactgatagg gacatctcta tcactgatag ggacttctct atcactgata gggaagtatg
    10381 ttctctctct tctcttctct ctctctttct cgaatgttct ctctcttctc ttctctctct
    10441 ctttctcgat ggccggggcg cgccaggttt cgactttcac ttttctctat cactgatagg
    10501 gagtggtaaa ctcgactttc acttttctct atcactgata gggagtggta aactcgactt
    10561 tcacttttct ctatcactga tagggagtgg taaactcgac tttcactttt ctctatcact
    10621 gatagggagt ggtaaactcg actttcactt ttctctatca ctgataggga gtggtaaact
    10681 cgactttcac ttttctctat cactgatagg gagtggtaaa ctcgactttc acttttctct
    10741 atcactgata gggagatccg agctcgtaaa ctcgactttc acttttctct atcactgata
    10801 gggagtggta aactcgactt tcacttttct ctatcactga tagggagtgg taaactcgac
    10861 tttcactttt ctctatcact gatagggagt ggtaaactcg aagcgccgga tccaaggttt
    10921 atgtgattca tataaatacg gccgcacaaa ctctcttcgg cattagtaca atacaaacaa
    10981 tcaaagtgaa aacattaaaa agaaataaca agcgatttaa attaagaaaa actacgttca
    11041 agtgaacaag aaattaaata aatcagaaag tgtacaaaag aaagtaaaat aattctataa
    11101 aaagtgaatt ttaagcaaca agttctataa atagaaaaac tattacaaat cattcatcaa
    11161 gtttatatac acaacaaaga agaaaaacaa agtcacacaa atgaaaaggc atgccaaaga
    11221 agaagcggaa ggtcggtatc cacggagtcc cagcagccga caagaagtac agcatcggcc
    11281 tggacatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac gaggtgccca
    11341 gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg
    11401 gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca
    11461 gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg
    11521 agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag
    11581 aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc
    11641 acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg
    11701 ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc
    11761 tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg
    11821 tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca
    11881 aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc
    11941 tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc ctgggcctga
    12001 cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg
    12061 acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc
    12121 tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga
    12181 acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc
    12241 accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag
    12301 agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc ggagccagcc
    12361 aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac
    12421 tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca
    12481 gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt
    12541 tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc
    12601 cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga
    12661 gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc gcttccgccc
    12721 agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc
    12781 ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc aaagtgaaat
    12841 acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg
    12901 tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact
    12961 tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg
    13021 cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca
    13081 atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca
    13141 gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga
    13201 agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg
    13261 gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg
    13321 ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc
    13381 agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg
    13441 gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga
    13501 aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga
    13561 ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca
    13621 aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg
    13681 agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg
    13741 acatcaaccg gctgtccgac tacgatgtgg accatatcgt gcctcagagc tttctgaagg
    13801 acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc aagagcgaca
    13861 acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg
    13921 ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga
    13981 gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa
    14041 agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc
    14101 tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg
    14161 atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc
    14221 tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg
    14281 tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa
    14341 tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg
    14401 agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa
    14461 ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca
    14521 tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt
    14581 ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta
    14641 agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag
    14701 tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca
    14761 tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag
    14821 aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg
    14881 gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc
    14941 cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc
    15001 ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga
    15061 tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca
    15121 aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata
    15181 tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca
    15241 ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc
    15301 accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgaca
    15361 aaaggccggc ggccacgaaa aaggccggcc aggcaaaaaa gaaaaagtaa tagaatgaat
    15421 cgtttttaaa ataacaaatc aattgtttta taatattcgt acgattcttt gattatgtaa
    15481 taaaatgtga tcattaggaa gattacgaaa aatataaaaa atatgagttc tgtgtgtata
    15541 acaaatgctg taaacgccac aattgtgttt gttgcaaata aacccatgat tatttgatta
    15601 aaattgttgt tttctttgtt catagacaat agtgtgtttt gcctaaacgt gtactgcata
    15661 aactccatgc gagtgtatag cgagctagtg gctaacgctt gccccaccaa agtagattcg
    15721 tcaaaatcct caatttcatc accctcctcc aagtttaaca tttggccgtc ggaattaact
    15781 tctaaagatg ccacataatc taataaatga aatagagatt caaacgtggc gtcatcgtcc
    15841 gtttcgacca tttccgaaaa gaactcgggc ataaactcta tgatttctct ggacgtggtg
    15901 ttgtcgaaac tctcaaagta cgcagtcagg aacgtgcgcg acatgtcgtc gggaaactcg
    15961 cgcggaaaca tgttgttgta accgaacggg tcccatagcg ccaaaaccaa atctgccagc
    16021 gtcaatagaa tgagcacgat gccgacaatg gagctggctt ggatagcgat tcgagttaac
    16081 tactgtccgt ttttatttgt gtaataaaat gaacggaacg ctgaattctg ga
//
