LOCUS       dCas9_1-3-5_FINA       16085 bp    DNA     circular SYN 05-NOV-2024
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16085)
  AUTHORS   NC State
  TITLE     Direct Submission
  JOURNAL   Exported Aug 18, 2025 from SnapGene 8.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..16085
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    265..364
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
                     /note="color: #31849b"
     misc_feature    293..334
                     /label=attP
                     /note="color: #ffff00"
     protein_bind    499..517
                     /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    520..538
                     /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    541..559
                     /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    562..580
                     /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    583..601
                     /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    604..622
                     /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    625..643
                     /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    646..664
                     /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    667..685
                     /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    688..706
                     /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    816..834
                     /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    866..884
                     /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    908..926
                     /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    950..968
                     /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    1000..1018
                     /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    1042..1060
                     /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    1092..1110
                     /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    1134..1422
                     /label=Lc Hsp70 promoter
                     /note="color: #a6acb3; direction: RIGHT"
     CDS             1426..1473
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /note="color: #993366"
                     /translation="KRPAATKKAGQAKKKK"
     CDS             1426..1473
                     /codon_start=1
                     /label=NLS
                     /note="color: #cc99ff"
                     /translation="KRPAATKKAGQAKKKK"
     CDS             join(1474..1497,1498..1500,1501..4056,4057..4059,
                     4060..5574)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the N863A mutation in the HNH 
                     catalytic domain"
                     /note="This feature has 5 segments:
                        1: 1474 .. 1497 / #993366
                        2: 1498 .. 1500 / #cc99b2
                        3: 1501 .. 4056 / #993366
                        4: 4057 .. 4059 / #cc99b2
                        5: 4060 .. 5574 / #993366"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKARGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     primer_bind     4212..4231
                     /label=dCas9 F
                     /note="color: black; sequence: CGGCTTCATCAAGAGACAGC; added:
                     2023-06-23"
     primer_bind     complement(4538..4557)
                     /label=dCas9 R
                     /note="color: black; sequence: GATTTCCTGCTCGCTCTTGG; added:
                     2023-06-23"
     CDS             5629..5649
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /note="color: #993366"
                     /translation="PKKKRKV"
     misc_feature    6524..6674
                     /label=Lc U6b terminator
                     /note="color: #a6acb3"
     misc_feature    complement(6675..7215)
                     /label=gRNA array
                     /note="color: #00ffff; direction: LEFT"
     primer_bind     6923..6945
                     /label=gRNA F
                     /note="color: black; sequence: ACATGATAAGCAGTTGTTGTGGT; 
                     added: 2023-06-23"
     primer_bind     complement(7157..7176)
                     /label=gRNA R
                     /note="color: black; sequence: AGTGGTAGAATGCTCGCCTG; added:
                     2023-06-23"
     misc_feature    complement(7216..7362)
                     /label=LcU6b promoter
                     /note="color: #a6acb3; direction: LEFT"
     misc_feature    7216..7362
                     /label=Lc U6b promoter
                     /note="color: #a6acb3"
     misc_feature    7385..7892
                     /label=Tub 3' UTR
                     /note="color: #ffcc00"
     primer_bind     7868..7887
                     /label=RFP F
                     /note="color: black; sequence: ATGATGGGTTTGGCTGCAAG; added:
                     2023-06-23"
     CDS             complement(7891..8568)
                     /codon_start=1
                     /label=RFPex
                     /note="color: #ff0000"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTFIYHVKFIGVNFPSDGLVMQKKTLGWEPSTERLYPRDGVLKGEIHK
                     ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     8325..8344
                     /label=F1
                     /note="color: black; sequence: AATCGGGAATATCAGCGGGA; added:
                     2024-11-05"
     primer_bind     complement(8565..8586)
                     /label=RFP R
                     /note="color: black; sequence: GCTCAGCTTAAAGGATCCATGG; 
                     added: 2023-06-23"
     misc_feature    complement(8569..10783)
                     /label=Lc Hsp83
                     /note="color: #a6acb3; direction: LEFT"
     protein_bind    10865..10964
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
                     /note="color: #31849b"
     misc_feature    10895..10936
                     /label=attP
                     /note="color: #ffff00"
     misc_feature    11823..12581
                     /label=3pBacmut
                     /note="color: #ff99cc"
     primer_bind     complement(12634..12650)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     promoter        13124..13228
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             join(13229..13297,13298..14089)
                     /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: 13229 .. 13297 / #ccffcc / signal sequence
                        2: 13298 .. 14089 / #ccffcc
                     Cleavage site after base 13297"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      14260..14848
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     protein_bind    15136..15157
                     /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(15172..15177,15178..15195,15196..15202)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This forward directional feature has 3 segments:
                        1: 15172 .. 15177 / #ffffff / -35
                        2: 15178 .. 15195 / #ffffff
                        3: 15196 .. 15202 / #ffffff / -10"
     protein_bind    15210..15226
                     /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     15234..15250
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     misc_feature    15306..15964
                     /label=5' pBac mut
                     /note="color: #ff99cc"
ORIGIN
        1 aacattcatg tgtgggtgat tacactcaca caaaaacaca cacattcaca caaacacaca
       61 cacttataaa gatgagaaca tttaacaaaa acaattataa aactgatggc aatggagcaa
      121 tgatgtgatg aagctgatga aagttttgaa tattcattca agagttaaaa acatgaacaa
      181 acatgcacaa aattagtcga gactgacgga cacaccgaag ccccggcggc aaccctcagc
      241 ggatgccccg gggcttcacg ttttcccagg tcagaagcgg ttttcgggag tagtgcccca
      301 actggggtaa cctttgagtt ctctcagttg ggggcgtagg gtcgccgaca tgacacaagg
      361 ggttgtgacc ggggtggaca cgtacgcggg tgcttacgac cgtcagtcgc gcgagcgcga
      421 ctagtacaag ccgaattctg cagatggcgc gccaacctat gcttatccgt ttttatttgt
      481 gtaatggccg cagaattctc tctatcactg atagggaagt ctctatcact gatagggacc
      541 tctctatcac tgatagggaa atctctatca ctgataggga tctctctatc actgataggg
      601 acttctctat cactgatagg gacgtctcta tcactgatag ggaactctct atcactgata
      661 gggacatctc tatcactgat agggacttct ctatcactga tagggaagta tgttctctct
      721 cttctcttct ctctctcttt ctcgaatgtt ctctctcttc tcttctctct ctctttctcg
      781 atggccgggg cgcgccaggt ttcgactttc acttttctct atcactgata gggagatccg
      841 agctcgtaaa ctcgactttc acttttctct atcactgata gggagtggta aactcgactt
      901 tcacttttct ctatcactga tagggagtgg taaactcgac tttcactttt ctctatcact
      961 gatagggaga tccgagctcg taaactcgac tttcactttt ctctatcact gatagggagt
     1021 ggtaaactcg actttcactt ttctctatca ctgataggga gatccgagct cgtaaactcg
     1081 actttcactt ttctctatca ctgataggga gtggtaaact cgaagcgccg gatccaaggt
     1141 ttatgtgatt catataaata cggccgcaca aactctcttc ggcattagta caatacaaac
     1201 aatcaaagtg aaaacattaa aaagaaataa caagcgattt aaattaagaa aaactacgtt
     1261 caagtgaaca agaaattaaa taaatcagaa agtgtacaaa agaaagtaaa ataattctat
     1321 aaaaagtgaa ttttaagcaa caagttctat aaatagaaaa actattacaa atcattcatc
     1381 aagtttatat acacaacaaa gaagaaaaac aaagtcacac aaatgaaaag gccggcggcc
     1441 acgaaaaagg ccggccaggc aaaaaagaaa aaggacaaga agtacagcat cggcctggcc
     1501 atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt gcccagcaag
     1561 aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct gatcggagcc
     1621 ctgctgttcg acagcggcga aacagccgag gccacccggc tgaagagaac cgccagaaga
     1681 agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag caacgagatg
     1741 gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt ggaagaggat
     1801 aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc ctaccacgag
     1861 aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga caaggccgac
     1921 ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca cttcctgatc
     1981 gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca gctggtgcag
     2041 acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc
     2101 atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc ccagctgccc
     2161 ggcgagaaga agaatggcct gttcggcaac ctgattgccc tgagcctggg cctgaccccc
     2221 aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag caaggacacc
     2281 tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc cgacctgttt
     2341 ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag agtgaacacc
     2401 gagatcacca aggcccccct gagcgcctct atgatcaaga gatacgacga gcaccaccag
     2461 gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta caaagagatt
     2521 ttcttcgacc agagcaagaa cggctacgcc ggctacattg acggcggagc cagccaggaa
     2581 gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga ggaactgctc
     2641 gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc
     2701 ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcagga agatttttac
     2761 ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg catcccctac
     2821 tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag aaagagcgag
     2881 gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgcttc cgcccagagc
     2941 ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt gctgcccaag
     3001 cacagcctgc tgtacgagta cttcaccgtg tataacgagc tgaccaaagt gaaatacgtg
     3061 accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaggc catcgtggac
     3121 ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag
     3181 aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc
     3241 ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct ggacaatgag
     3301 gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga ggacagagag
     3361 atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt gatgaagcag
     3421 ctgaagcggc ggagatacac cggctggggc aggctgagcc ggaagctgat caacggcatc
     3481 cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg cttcgccaac
     3541 agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga catccagaaa
     3601 gcccaggtgt ccggccaggg cgatagcctg cacgagcaca ttgccaatct ggccggcagc
     3661 cccgccatta agaagggcat cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg
     3721 atgggccggc acaagcccga gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc
     3781 cagaagggac agaagaacag ccgcgagaga atgaagcgga tcgaagaggg catcaaagag
     3841 ctgggcagcc agatcctgaa agaacacccc gtggaaaaca cccagctgca gaacgagaag
     3901 ctgtacctgt actacctgca gaatgggcgg gatatgtacg tggaccagga actggacatc
     3961 aaccggctgt ccgactacga tgtggaccac atcgtgcctc agagctttct gaaggacgac
     4021 tccatcgaca acaaggtgct gaccagaagc gacaaggccc ggggcaagag cgacaacgtg
     4081 ccctccgaag aggtcgtgaa gaagatgaag aactactggc ggcagctgct gaacgccaag
     4141 ctgattaccc agagaaagtt cgacaatctg accaaggccg agagaggcgg cctgagcgaa
     4201 ctggataagg ccggcttcat caagagacag ctggtggaaa cccggcagat cacaaagcac
     4261 gtggcacaga tcctggactc ccggatgaac actaagtacg acgagaatga caagctgatc
     4321 cgggaagtga aagtgatcac cctgaagtcc aagctggtgt ccgatttccg gaaggatttc
     4381 cagttttaca aagtgcgcga gatcaacaac taccaccacg cccacgacgc ctacctgaac
     4441 gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc tggaaagcga gttcgtgtac
     4501 ggcgactaca aggtgtacga cgtgcggaag atgatcgcca agagcgagca ggaaatcggc
     4561 aaggctaccg ccaagtactt cttctacagc aacatcatga actttttcaa gaccgagatt
     4621 accctggcca acggcgagat ccggaagcgg cctctgatcg agacaaacgg cgaaaccggg
     4681 gagatcgtgt gggataaggg ccgggatttt gccaccgtgc ggaaagtgct gagcatgccc
     4741 caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa agagtctatc
     4801 ctgcccaaga ggaacagcga taagctgatc gccagaaaga aggactggga ccctaagaag
     4861 tacggcggct tcgacagccc caccgtggcc tattctgtgc tggtggtggc caaagtggaa
     4921 aagggcaagt ccaagaaact gaagagtgtg aaagagctgc tggggatcac catcatggaa
     4981 agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta caaagaagtg
     5041 aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga aaacggccgg
     5101 aagagaatgc tggcctctgc cggcgaactg cagaagggaa acgaactggc cctgccctcc
     5161 aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg ctcccccgag
     5221 gataatgagc agaaacagct gtttgtggaa cagcacaagc actacctgga cgagatcatc
     5281 gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct ggacaaagtg
     5341 ctgtccgcct acaacaagca ccgggataag cccatcagag agcaggccga gaatatcatc
     5401 cacctgttta ccctgaccaa tctgggagcc cctgccgcct tcaagtactt tgacaccacc
     5461 atcgaccgga agaggtacac cagcaccaaa gaggtgctgg acgccaccct gatccaccag
     5521 agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg cgacagcgct
     5581 ggaggaggtg gaagcggagg aggaggaagc ggaggaggag gtagcggacc taagaaaaag
     5641 aggaaggtgt aaatagaatg aatcgttttt aaaataacaa atcaattgtt ttataatatt
     5701 cgtacgattc tttgattatg taataaaatg tgatcattag gaagattacg aaaaatataa
     5761 aaaatatgag ttctgtgtgt ataacaaatg ctgtaaacgc cacaattgtg tttgttgcaa
     5821 ataaacccat gattatttga ttaaaattgt tgttttcttt gttcatagac aatagtgtgt
     5881 tttgcctaaa cgtgtactgc ataaactcca tgcgagtgta tagcgagcta gtggctaacg
     5941 cttgccccac caaagtagat tcgtcaaaat cctcaatttc atcaccctcc tccaagttta
     6001 acatttggcc gtcggaatta acttctaaag atgccacata atctaataaa tgaaatagag
     6061 attcaaacgt ggcgtcatcg tccgtttcga ccatttccga aaagaactcg ggcataaact
     6121 ctatgatttc tctggacgtg gtgttgtcga aactctcaaa gtacgcagtc aggaacgtgc
     6181 gcgacatgtc gtcgggaaac tcgcgcggaa acatgttgtt gtaaccgaac gggtcccata
     6241 gcgccaaaac caaatctgcc agcgtcaata gaatgagcac gatgccgaca atggagctgg
     6301 cttggatagc gattcgagtt aactactgtc cgtttttatt tgtgtaataa aatgaacgga
     6361 acgctgaatt ctggaacatt agaaattctt tgttatagaa tctataaatt tatgtcaaag
     6421 ttttgtcgtt tcttatgcga tgagagatct taaaaggcaa gtaaattgag ctgtcctatt
     6481 caaacatttt tctattaaat tgttggtacc ttccgcgtcg acgattcaat atcgagcaaa
     6541 aatacgaacg catatgattc catgaaacaa atgtatttct ttagacgttt ctaaaaccaa
     6601 ttccgacaat tatttgattc tatgacaact ccaaatagtt ttgtttatta aaaaataaaa
     6661 tttcaacaaa aaaagcaccg actcggtgcc actttttcaa gttgataacg gactagcctt
     6721 attttaactt gctatttcta gctctaaaac tcacctaaac atagagataa tgcaccagcc
     6781 gggaatcgaa cccgggtctg taccgtggca gggtactatt ctaccactag accactggtg
     6841 ctttgttagc accgactcgg tgccactttt tcaagttgat aacggactag ccttatttta
     6901 acttgctatt tctagctcta aaacatgata agcagttgtt gtggtgcacc agccgggaat
     6961 cgaacccggg tctgtaccgt ggcagggtac tattctacca ctagaccact ggtgctttgt
     7021 tagcaccgac tcggtgccac tttttcaagt tgataacgga ctagccttat tttaacttgc
     7081 tatttctagc tctaaaacgc taagttttca actacgtatg catcggccgg gaatcgaacc
     7141 cgggccgccc gcgtggcagg cgagcattct accactgaac caccgatgct tgatgtctac
     7201 acacactcaa agcccaaaga tcatcacttt aataaactat ttaaccgttc tttttgagaa
     7261 aaaagaatat gggaattata aacaacaata ctatatgttg aaaacataaa aatatcattt
     7321 acagattagg gtttgttatt atcacattat tactacctga gtcgtcgacg ccggatcctt
     7381 caagttcctc ttgtacccat caaaatgagg ccaaactagt gctgttttta atgcactcac
     7441 tatgcatttt tatagcttcc tctacacctc ggcaagttca ttaaactttt gccaagcatt
     7501 ttaacaatta ctatttttag ttgttttccc tctttttccc cccattaagt aataaaacat
     7561 taaaaaattt tcttttggaa tattttcatt caatgcagct taagcctttc attttattag
     7621 gttcttgttt ttttcgtgtt tttaaagtgc acttctttaa aatttgttct tttcattcat
     7681 gttttttcaa atttgaaaat aaaaaaaatt tacaagtgtt ttgttttgga ttcgttatca
     7741 tttttaaaat aacagtttgt ttaaaaaaaa gtatattgaa tgtttagttg agtagtttca
     7801 gctaataggt cgagaatttt ttcaatacac acttcttctg tctttttttt ggttttgatg
     7861 gttgtcgatg atgggtttgg ctgcaagctt ttattgaaac aagtggtgac gggcttcggc
     7921 acgttcgtat tgttctacta cggtgtaatc ttcattatga ctggtaatat ccaattttga
     7981 atcgacgtag tagtaacctg gcaacttaac gggctttttg gccatgtaga tggacttaaa
     8041 ttcaaccaag tagtgaccac cacccttcaa tttcaaagcc ttatgaattt cacctttcaa
     8101 gacaccatca cggggataca aacgttcagt agaaggttcc caacccaagg ttttcttttg
     8161 cataacaaga ccatctgatg gaaagtttac accgatgaat ttgacgtggt aaataaaagt
     8221 accatcttgc aaagaggaat cttgagtaac ggtgacaaca ccaccatctt cgaaattcat
     8281 tacacgttcc cacttaaaac cttcggggaa ggacaacttt ttataatcgg gaatatcagc
     8341 gggatgctta acgtatacct tagaaccgta ttggaattgt ggggacaaaa tatcccaggc
     8401 gaatggcaag ggaccaccct tggtaacttg caatttagca gtttgagtac cttcgtaggg
     8461 tttaccttca ccttcacctt cgatttcaaa ttcgtgacca ttgacagaac cttccatgtg
     8521 taccttaaaa cgcataaaag gcttgataac attttcagta gaatccatgg atcctttaag
     8581 ctgagcaatt tcagcttgga aagcaaaagt ttcaacttct tcaggcatct gtgtttaata
     8641 aaaaaaaaat agaaaaatac aaattattag taaaataatc aatcaaaata tgaagtataa
     8701 ataattttaa tgacgcacgt ttttcgttaa gtgtagggag tgggattggt actgaaaatt
     8761 tttaataaaa atctagaaat ctctttaagt taagaattac tagtttttta ggggtggatt
     8821 gggtatgctt tttaacgaat gaaaacatga aatatcgatt ataatttcga aaaaattttg
     8881 aaaaattttc tttaaaaaat cgcatgctag aaatttcttt ttattagcat ggtgaaattt
     8941 tctttatata ctagatttat tttctaaaaa tgtcatggcc aaagaaaatt taatctagaa
     9001 caatttacaa ttacttagta acgttattct taaaccatat tgaaggtttt tctttctggc
     9061 aaattacttg agtttttaca tcacactttt ttcaaaagaa caaaaacgtt ttgttactac
     9121 attttatttt ttttaataaa aacttacctt gatgtgtttg ttttatatta atttcacaaa
     9181 aataaatact ttttaaatta tttttctttt tgcaattgaa attaaacaac tacacacttt
     9241 tttaatgttc ttttttcaga cttttcttta ttaagaaaat taaaacactt ttaactttct
     9301 cccgtttgga attcttggtg ttttttgcac aatttttctt tcaagactga tgccggttag
     9361 cagccatgca ctgaatttat aggtaaaaca aaaataaacg aaaatactag aatgttctgg
     9421 aacaaactac ccgtacttcc gtaggctcta aacaacatca tgtggttgtt tatttggtga
     9481 taacaaaaga aattggttac attttagata tataaaaaaa ttaatttatt aaaattgtag
     9541 aaaaccaata ttaattatag gtttttattg ttattttaat ttattagttg caaattatca
     9601 aaaaaatgtg caaaacattc aacatgatta atgcccacac agcgttgtat atatgtagaa
     9661 gcagtggctt ttaaactgta aaattgcatc aatgtcaaaa tttctacaat gttctctctg
     9721 tttttaataa catgcggaat gaatcaacta taatttactt tatttattta tgcaaaaata
     9781 tataaaagtt attaattaaa tgaaaaagga attaatctta ggcattgctg agtaatgtca
     9841 tgacataaac taaatatttt tctagtcaaa caatgaactt atttatttca aaaacgtttt
     9901 attctttaaa ttatagttaa aaagtcttat taattgacaa atctatgtgt aattaagtgt
     9961 tttttgaact aagttacaag cttaatatgt atttgattaa ttgaaatatg tattaactat
    10021 taatttttaa attaaataaa tttaatatta attatatatt aaactttgtg ctttatatgg
    10081 aaatagtatg tatgatttaa tcacattcga tttacatatt agtcatcata aattataaat
    10141 gaaagtaatt taattgtttt attaaagaaa ataataacat tattaaaaca ataatattgc
    10201 tatcaaaatt tataaaaatg cataaaatga aaaaatattg attttctctc atcatggggg
    10261 cattaataat tttttcgcca gtgcagctga tgtgcagtaa aacatcagag ttgccggtct
    10321 tgaataaaaa caatgaaaag tggaataatt taaaaatgtg acaatcatat acttttaaca
    10381 tattttataa aatttgactg taaaaattta aaaattaaaa tttgcaaaca ttatctcata
    10441 catattataa caactctttt gtttttatat aaatttaata aatgaaatca aaaattctct
    10501 cacaaatttg taatttccaa aagtgtggcc ttttattttc ccctttttag gtaaaaattt
    10561 caaatatcta caaccctgta aatgacagct gtttgcgctc ttttatgtat tttttttgtt
    10621 tatcatgtcg gttttttagt tcgcggtgca atttactcga aatttcttaa agtttgttga
    10681 atttttggcg gtataattat aatattttgt tttagtgttc tttttatcaa attataatgt
    10741 ttaagtgaat tattttctga taatggtttc aacacgtggg cccatctgca gaattcggct
    10801 tgtactagtc gcgctcgcgc gactgacggt cgtaagcacc cgcgtacgtg tccaccccgg
    10861 tcacaacccc ttgtgtcatg tcggcgaccc tacgccccca actgagagaa ctcaaaggtt
    10921 accccagttg gggcactact cccgaaaacc gcttctgacc tgggaaaacg tgaagccccg
    10981 gggcatccgc tgagggttgc cgccggggct tcggtgtgtc cgtcagtgag ctcctcaggt
    11041 aataataaca actacaacca aaaaacaaaa gcaacaacaa cgacaacttc tacagggcaa
    11101 gtaacaaaac atagtatttg tcaacgacac actatatgaa tgatgatgat gatgcaacaa
    11161 cagcaacaac agcaacaaca gctcgagtat caacaacaca acaaaaaaaa cttttttaaa
    11221 aagacacagc tgctggcaac aaaataccac aaaagatata aactaacgca gttgccacac
    11281 acacacatac atacataaat taaaatatta tcaaacacac acatatacat atatcacaac
    11341 atacctatat acaaaaccac atttactttg ataaaaggcg attcaacaaa aaagtagtcg
    11401 tcatcctcct tcatttatcc ttgcatttca cgtgtatctg tgtgtgttat aaaacctgct
    11461 ggccaaaagg caggtaggta gttgacaaca caaatatttt acttcttttt catcatcagc
    11521 atcaacaaca gcaggagcaa catattgtag acacacactc tctgtttctt cacttcacac
    11581 acatacaagt aaattatgac agtgaaaaag acgggtattt ccaagcttca gtttcgttta
    11641 ctttaggtca ttaagatgac tgtcctgata gcttgatatc aatggaatca acgggattga
    11701 taaatggaat tactgtgatt gataaatagc ttcgaaagac ctcgttactt cagggtactt
    11761 aacaaaattt actgttggcc gcgaaccgcg ccatggcgcc ggccgggagc cggcgcatgc
    11821 gaaggtcgcc tatggcatta ttgtacggaa tgataaacat tgcctgcata aattctttta
    11881 ttatatacag ccataatgtc agtagcaacg gagaaaaggt tcaaagtcgc aaaaaattta
    11941 tgagaaacct ttacatgagc ctgacgtcat cgtttatgcg taagcgttta gaagctccta
    12001 ctttgaagag atatttgcgc gataatatct ctaatatttt gccaaatgaa gtgcctggta
    12061 catcagatga cagtactgaa gagccagtaa tgaaaaaacg tacttactgt acttactgcc
    12121 cctctaaaat aaggcgaaag gcaaatgcat cgtgcaaaaa atgcaaaaaa gttatttgtc
    12181 gagagcataa tattgatatg tgccaaagtt gtttctgact gactaataag tataatttgt
    12241 ttctattatg tataagttaa gctaattact tattttataa tacaacatga ctgtttttaa
    12301 agtacaaaat aagtttattt ttgtaaaaga gagaatgttt aaaagttttg ttactttata
    12361 gaagaaattt tgagtttttg ttttttttta ataaataaat aaacataaat aaattgtttg
    12421 ttgaatttat tattagtatg taagtgtaaa tataataaaa cttaatatct attcaaatta
    12481 ataaataaac ctcgatatac agaccgataa aacacatgcg tcaattttac gcatgattat
    12541 ctttaacgta cgtcacaata tgattatctt tctagggtta agggcccggg atccgatatc
    12601 tagatgcatt cgcgaggtac cgagctcgaa ttcactggcc gtcgttttac aacgtcgtga
    12661 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
    12721 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
    12781 tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
    12841 catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
    12901 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
    12961 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
    13021 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
    13081 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
    13141 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
    13201 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
    13261 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
    13321 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
    13381 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
    13441 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
    13501 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
    13561 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
    13621 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
    13681 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
    13741 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
    13801 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
    13861 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
    13921 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
    13981 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
    14041 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
    14101 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
    14161 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
    14221 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
    14281 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
    14341 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
    14401 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
    14461 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
    14521 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
    14581 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
    14641 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
    14701 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
    14761 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
    14821 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
    14881 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
    14941 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
    15001 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
    15061 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
    15121 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
    15181 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
    15241 cagctatgac catgattacg ccaagcttgc atgcaggcct ctgcagtcga cgggcccaag
    15301 cttcctttaa ccctagaaag atagtctgcg taaaattgac gcatgcattc ttgaaatatt
    15361 gctctctctt tctaaatagc gcgaatccgt cgctgtgcat ttaggacatc tcagtcgccg
    15421 cttggagctc ccgtgaggcg tgcttgtcaa tgcggtaagt gtcactgatt ttgaactata
    15481 acgaccgcgt gagtcaaaat gacgcatgat tatcttttac gtgactttta agatttaact
    15541 catacgataa ttatattgtt atttcatgtt ctacttacgt gataacttat tatatatata
    15601 ttttcttgtt atagatatcg tgactaatat ataataaaat gggtagttct ttagacgatg
    15661 agcatatcct ctctgctctt ctgcaaagcg atgacgagct tgttggtgag gattctgaca
    15721 gtgaaatatc agatcacgta agtgaagatg acgtccagag cgatacagaa gaagcgttta
    15781 tagatgaggt acatgaagtg cagccaacgt caagcggtag tgaaatatta gacgaacaaa
    15841 atgttattga acaaccaggt tcttcattgg cttctaacag aatcttgacc ttgcgacaga
    15901 ggactattag aggtaagaat aaacattgtt ggtcaacttc aaagtccacg aggcgtagcg
    15961 gagtggtacc aggccctcga gtctgggccc atgcatggga gctctcccaa gatcttatgg
    16021 tcgacctgca ggcaacattc atgtgtgggt gattacactc acacaaaaac acacacattc
    16081 acaca
//
