LOCUS       sequence-335585-       10764 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gal inducible expression of D10A-Cas9 nickase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10764)
  TITLE     Burgess Lab Plasmids
REFERENCE   2  (bases 1 to 10764)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pITI2
FEATURES             Location/Qualifiers
     source          1..10764
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    1943..2446
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     2488..2506
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2544..2566)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2666..2685
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(3179..4273)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(4274..4681)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(4982..5437)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5119..5140)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5331..5350
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5563..5585
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5577..5594
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5578..5594
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5604..5623
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5604..5622
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(5641..5888)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     5854..5872
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(5904..5924)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(join(5937..10010,10011..10013,10014..10040))
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(D10A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the D10A mutation in the RuvC catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     primer_bind     complement(10073..10096)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(10505..10525)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(10505..10523)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(10544..10560)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(10544..10560)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(10557..10579)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    10568..10584
                     /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(join(10592..10598,10599..10616,10617..10622))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10637..10658
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc tgggtccttt tcatcacgtg ctataaaaat aattataatt taaatttttt
     1981 aatataaata tataaattaa aaatagaaag taaaaaaaga aattaaagaa aaaatagttt
     2041 ttgttttccg aagatgtaaa agactctagg gggatcgcca acaaatacta ccttttatct
     2101 tgctcttcct gctctcaggt attaatgccg aattgtttca tcttgtctgt gtagaagacc
     2161 acacacgaaa atcctgtgat tttacatttt acttatcgtt aatcgaatgt atatctattt
     2221 aatctgcttt tcttgtctaa taaatatata tgtaaagtac gctttttgtt gaaatttttt
     2281 aaacctttgt ttattttttt ttcttcattc cgtaactctt ctaccttctt tatttacttt
     2341 ctaaaatcca aatacaaaac ataaaaataa ataaacacag agtaaattcc caaattattc
     2401 catcattaaa agatacgagg cgcgtgtaag ttacaggcaa gcgatccgtc ctaagaaacc
     2461 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     2521 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     2581 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     2641 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     2701 atcgactacg tcgttaaggc cgtttctgac agagtaaaat tcttgaggga actttcacca
     2761 ttatgggaaa tggttcaaga aggtattgac ttaaactcca tcaaatggtc aggtcattga
     2821 gtgtttttta tttgttgtat tttttttttt ttagagaaaa tcctccaata tataaattag
     2881 gaatcatagt ttcatgattt tctgttacac ctaacttttt gtgtggtgcc ctcctccttg
     2941 tcaatattaa tgttaaagtg caattctttt tccttatcac gttgagccat tagtatcaat
     3001 ttgcttacct gtattccttt acatcctcct ttttctcctt cttgataaat gtatgtagat
     3061 tgcgtatata gtttcgtcta ccctatgaac atattccatt ttgtaatttc gtgtcgtttc
     3121 tattatgaat ttcatttata aagtttatgt acaaatatca taaaaaaaga gaatcttttt
     3181 aagcaaggat tttcttaact tcttcggcga cagcatcacc gacttcggtg gtactgttgg
     3241 aaccacctaa atcaccagtt ctgatacctg catccaaaac ctttttaact gcatcttcaa
     3301 tggccttacc ttcttcaggc aagttcaatg acaatttcaa catcattgca gcagacaaga
     3361 tagtggcgat agggttgacc ttattctttg gcaaatctgg agcagaaccg tggcatggtt
     3421 cgtacaaacc aaatgcggtg ttcttgtctg gcaaagaggc caaggacgca gatggcaaca
     3481 aacccaagga acctgggata acggaggctt catcggagat gatatcacca aacatgttgc
     3541 tggtgattat aataccattt aggtgggttg ggttcttaac taggatcatg gcggcagaat
     3601 caatcaattg atgttgaacc ttcaatgtag ggaattcgtt cttgatggtt tcctccacag
     3661 tttttctcca taatcttgaa gaggccaaaa cattagcttt atccaaggac caaataggca
     3721 atggtggctc atgttgtagg gccatgaaag cggccattct tgtgattctt tgcacttctg
     3781 gaacggtgta ttgttcacta tcccaagcga caccatcacc atcgtcttcc tttctcttac
     3841 caaagtaaat acctcccact aattctctga caacaacgaa gtcagtacct ttagcaaatt
     3901 gtggcttgat tggagataag tctaaaagag agtcggatgc aaagttacat ggtcttaagt
     3961 tggcgtacaa ttgaagttct ttacggattt ttagtaaacc ttgttcaggt ctaacactac
     4021 cggtacccca tttaggacca cccacagcac ctaacaaaac ggcatcagcc ttcttggagg
     4081 cttccagcgc ctcatctgga agtggaacac ctgtagcatc gatagcagca ccaccaatta
     4141 aatgattttc gaaatcgaac ttgacattgg aacgaacatc agaaatagct ttaagaacct
     4201 taatggcttc ggctgtgatt tcttgaccaa cgtggtcacc tggcaaaacg acgatcttct
     4261 taggggcaga cattagaatg gtatatcctt gaaatatata tatatatatt gctgaaatgt
     4321 aaaaggtaag aaaagttaga aagtaagacg attgctaacc acctattgga aaaaacaata
     4381 ggtccttaaa taatattgtc aacttcaagt attgtgatgc aagcatttag tcatgaacgc
     4441 ttctctattc tatatgaaaa gccggttccg gcgctctcac ctttcctttt tctcccaatt
     4501 tttcagttga aaaaggtata tgcgtcaggc gacctctgaa attaacaaaa aatttccagt
     4561 catcgaattt gattctgtgc gatagcgccc ctgtgtgttc tcgttatgtt gaggaaaaaa
     4621 ataatggttg ctaagagatt cgaactcttg catcttacga tacctgagta ttcccacagt
     4681 taactgcggt caagatattt cttgaatcag gcgccttaga ccgctcggcc aaacaaccaa
     4741 ttacttgttg agaaatagag tataattatc ctataaatat aacgtttttg aacacacatg
     4801 aacaaggaag tacaggacaa ttgattttga agagaatgtg gattttgatg taattgttgg
     4861 gattccattt ttaataaggc aataatatta ggtatgtaga tatactagaa gttctcctcg
     4921 accggtcgat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     4981 gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     5041 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     5101 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     5161 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     5221 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     5281 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     5341 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     5401 acacccgccg cgcttaatgc gccgctacag ggcgcgtcca ttcgccattc aggctgcgca
     5461 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     5521 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     5581 aaacgacggc cagtgagcgc gcgtaatacg actcactata gggcgaattg ggtaccggcc
     5641 gcaaattaaa gccttcgagc gtcccaaaac cttctcaagc aaggttttca gtataatgtt
     5701 acatgcgtac acgcgtctgt acagaaaaaa aagaaaaatt tgaaatataa ataacgttct
     5761 taatactaac ataactataa aaaaataaat agggacctag acttcaggtt gtctaactcc
     5821 ttccttttcg gttagagcgg atgtgggggg agggcgtgaa tgtaagcgtg acataactaa
     5881 ttacatgact cgagaagaga tcacaccttc ctcttcttct tggggtcagc cctgctgtct
     5941 ccaccgagct gagagaggtc gattcttgtt tcatagagcc ccgtaattga ctgatgaatc
     6001 agtgtggcgt ccaggacctc ctttgtagag gtgtaccgct ttctgtctat ggtggtgtcg
     6061 aagtacttga aggctgcagg cgcgcccaag ttggtcagag taaacaagtg gataatgttt
     6121 tctgcctgct ccctgatggg cttatccctg tgcttattgt aagcagaaag caccttatcg
     6181 aggttagcgt cggcgaggat cactcttttg gagaattcgc ttatttgctc gatgatctca
     6241 tcaaggtagt gtttgtgttg ttccacgaac agctgcttct gctcattatc ttcgggagac
     6301 cctttgagct tttcatagtg gctggccaga tacaagaaat taacgtattt agagggcagt
     6361 gccagctcgt tacctttctg cagctcgccc gcactagcga gcattcgttt ccggccgttt
     6421 tcaagctcaa agagagagta cttgggaagc ttaatgatga ggtctttttt gacctcttta
     6481 tatcctttcg cctcgagaaa gtcgatgggg tttttttcga agcttgatcg ctccatgatt
     6541 gtgatgccca gcagttcctt gacgcttttg agttttttag acttcccttt ctccactttg
     6601 gccacaacca gtacactgta agcgactgta ggagaatcga atccgccgta tttcttgggg
     6661 tcccaatctt ttttgcgtgc gatcagcttg tcgctgttcc ttttcgggag gatactttcc
     6721 ttggagaagc ctccggtctg tacttcggtc tttttaacga tgttcacctg cggcatggac
     6781 aggaccttcc ggactgtcgc gaaatcccta cccttgtccc acacgatttc tcctgtttct
     6841 ccgtttgttt cgataagtgg tcgcttccga atctctccat tggccagtgt aatctcggtc
     6901 ttgaaaaaat tcataatatt gctgtaaaag aagtacttag cggtggcctt gcctatttcc
     6961 tgctcagact ttgcgatcat tttcctaaca tcgtacactt tatagtctcc gtaaacaaat
     7021 tcagattcaa gcttgggata ttttttgata agtgcagtgc ctaccactgc attcaggtag
     7081 gcatcatgcg catggtggta attgttgatc tctctcacct tataaaactg aaagtccttt
     7141 ctgaaatctg agaccagctt agacttcaga gtaataactt tcacctctcg aatcagtttg
     7201 tcattttcat cgtacttggt gttcatgcgt gaatcgagaa tttgggccac gtgcttggtg
     7261 atctggcgtg tctcaacaag ctgccttttg atgaagccgg ctttatccaa ctcagacagg
     7321 ccacctcgtt cagccttagt cagattatcg aacttccgtt gtgtgatcag tttggcgttc
     7381 agcagctgcc gccaataatt tttcattttc ttgacaactt cttctgaggg gacgttatca
     7441 ctcttccctc tatttttatc ggatcttgtc aacactttat tatcaataga atcatctttg
     7501 agaaaagact ggggcacgat atgatccacg tcgtagtcgg agagccgatt gatgtccagt
     7561 tcctgatcca cgtacatgtc cctgccgttc tgcaggtagt acaggtagag cttctcattc
     7621 tgaagctggg tgttttcaac tgggtgttcc ttaaggattt gggaccccag ttcttttata
     7681 ccctcttcaa tcctcttcat cctttcccta ctgttcttct gtcccttctg ggtagtttgg
     7741 ttctctcggg ccatctcgat aacgatattc tcgggcttat gccttcccat tactttgacg
     7801 agttcatcca cgaccttaac ggtctgcagt attccctttt tgatagctgg gctacctgca
     7861 agattagcga tgtgctcgtg aagactgtcc ccctggccag aaacttgtgc tttctggatg
     7921 tcctccttaa aggtgagaga gtcatcatgg atcaactgca tgaagttccg gttggcaaat
     7981 ccatcggact taagaaaatc caggattgtc tttccactct gcttgtctcg gatcccattg
     8041 atcagttttc ttgacagccg cccccatcct gtatatcggc gcctcttgag ctgtttcatg
     8101 actttgtcgt cgaagagatg agcgtaagtt ttcaagcgtt cttcaatcat ctccctatct
     8161 tcaaacaacg taagggtgag gacaatgtcc tcaagaatgt cctcgttctc ctcattgtcc
     8221 aggaagtcct tgtctttaat gattttcagg agatcgtgat acgttcccag ggatgcgttg
     8281 aagcgatcct ccactccgct gatttcaaca gagtcgaaac attcaatctt tttgaaatag
     8341 tcttctttga gctgtttcac ggtaactttc cggttcgtct tgaagaggag gtccacgata
     8401 gctttcttct gctctccaga caggaatgct ggctttctca tcccttctgt gacgtatttg
     8461 accttggtga gctcgttata aactgtgaag tactcgtaca gcagagagtg tttaggaagc
     8521 accttttcgt taggcagatt tttatcaaag ttagtcatcc tttcgatgaa ggactgggca
     8581 gaggccccct tatccacgac ttcctcgaag ttccagggag tgatggtctc ttctgatttg
     8641 cgagtcatcc acgcgaatct ggaatttccc cgggcgaggg ggcctacata gtagggtatc
     8701 cgaaatgtga ggattttctc aatcttttcc ctgttatctt tcaaaaaggg gtagaaatcc
     8761 tcttgccgcc tgaggatagc gtgcagttcg cccaggtgaa tctggtgggg gatgcttcca
     8821 ttgtcgaaag tgcgctgttt gcgcaacaga tcttctctgt taagctttac cagcagctcc
     8881 tcggtgccgt ccattttttc caagatgggc ttaataaatt tgtaaaattc ctcctggctt
     8941 gctccgccgt caatgtatcc ggcgtagcca tttttagact gatcgaagaa aatttccttg
     9001 tacttctcag gcagttgctg tctgacaagg gccttcagca aagtcaagtc ttggtggtgc
     9061 tcatcatagc gcttgatcat actagcgctc agcggagctt tggtgatctc cgtgttcact
     9121 cgcagaatat cactcagcag aatggcgtct gacaggttct ttgccgccaa aaaaaggtct
     9181 gcgtactggt cgccgatctg ggccagcaga ttgtcgagat catcatcgta ggtgtctttg
     9241 ctcagttgaa gcttggcatc ttcggccagg tcgaagttag atttaaagtt gggggtcagc
     9301 ccgagtgaca gggcgataag attaccaaac aggccgttct tcttctcccc agggagctgt
     9361 gcgatgaggt tttcgagccg ccgggatttg gacagcctag cgctcaggat tgctttggcg
     9421 tcaactccgg atgcgttgat cgggttctct tcgaaaagct gattgtaagt ctgaaccagt
     9481 tggataaaga gtttgtcgac atcgctgttg tctgggttca ggtccccctc gatgaggaag
     9541 tgtccccgaa atttgatcat atgcgccagc gcgagataga tcaaccgcaa gtcagcctta
     9601 tcagtactgt ctacaagctt cttcctcaga tgatatatgg ttgggtactt ttcatggtac
     9661 gccacctcgt ccacgatatt gccaaagatt gggtggcgct cgtgcttttt atcctcctcc
     9721 accaaaaagg actcctccag cctatggaag aaagagtcat ccaccttagc catctcatta
     9781 ctaaagatct cctgcaggta gcagattctg ttctttcttc tggtgtatct tcttctagcg
     9841 gttcttttca atctagtagc ttcagcagtt tcaccagaat cgaacaacaa ggcaccaatc
     9901 aagttctttt tgatggagtg tctatcagtg ttacccaaaa ccttgaactt cttggatgga
     9961 accttgtatt catcagtaat aacagcccaa ccaacagagt ttgtaccgat agccaaacca
    10021 atagagtatt tcttatccat actagttcta gaatccgggg ttttttctcc ttgacgttaa
    10081 agtatagagg tatattaaca attttttgtt gatactttta ttacatttga ataagaagta
    10141 atacaaaccg aaaatgttga aagtattagt taaagtggtt atgcagtttt tgcatttata
    10201 tatccgttaa tagatcaaaa atcatcgctt cgctgattaa ttaccccaga aataaggcta
    10261 aaaaactaat cgcattatca tcctatggtt gttaatttga ttcgttcatt tgaaggtttg
    10321 tggggccagg ttactgccaa tttttcctct tcataaccat aaaagctagt attgtagaat
    10381 ctttattgtt cggagcagtg cggcgcgagg cacatctgcg tttcaggaac gcgaccggtg
    10441 aagacgagga cgcacggagg agagtcttcc ttcggagggc tgtcacgagc tccagctttt
    10501 gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg
    10561 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
    10621 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
    10681 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
    10741 gaggcggttt gcgtattggg cgct
//
