LOCUS       sequence-474294-       12007 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Combined sgRNA/Cas9 pLSB vector with hphMX6 (hygromycin) marker. 
            Golden Gate-ready for cloning custom sgRNA sequences..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12007)
  AUTHORS   Torres-Garcia S, Di Pompeo L, Eivers L, Gaborieau B, White SA, 
            Pidoux AL, Kanigowska P, Yaseen I, Cai Y, Allshire RC
  TITLE     SpEDIT: A fast and efficient CRISPR/Cas9 method for fission yeast.
  JOURNAL   Wellcome Open Res. 2020 Nov 24;5:274. doi: 
            10.12688/wellcomeopenres.16405.1. eCollection 2020.
  PUBMED    33313420
REFERENCE   2  (bases 1 to 12007)
  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"; journalName: "Wellcome 
            Open Res."; date: "2020-11-24"; volume: "5:274. doi"; pages: " 
            10.12688/wellcomeopenres.16405.1. eCollection 2020"
COMMENT     Sequence Label: pLSB-HYG
FEATURES             Location/Qualifiers
     source          1..12007
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     175..197
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     190..206
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     210..226
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             644..655
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             662..1375
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1387..1458
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1474..1501
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_RNA        1515..1590
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      1600..1619
                     /label=SUP4 terminator
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     primer_bind     complement(1620..1636)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1640..1656)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1653..1675)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1664..1680
                     /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(1688..1694,1695..1712,1713..1718))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1733..1754
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1871..1888)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2042..2630)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2122..2141)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2801..3592,3593..3661))
                     /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     3424..3443
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3662..3766)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3799..4540
                     /label=adh1 promoter
                     /note="promoter for the Schizosaccharomyces pombe adh1 gene
                     encoding alcohol dehydrogenase"
     CDS             4561..8664
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             4561..8664
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             8677..8697
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      8715..8962
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(8731..8749)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     9009..9027
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(9065..9087)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     9187..9206
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      9577..10358
                     /label=ars1
                     /note="Schizosaccharomyces pombe autonomously replicating 
                     sequence ars1"
     promoter        10432..10775
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             10776..11804
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MGKKPELTATSVEKFLIEKFDSVSDLMRLSEGEESRAFSFDVGGR
                     GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL
                     PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW
                     QTVMDDTVGASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM
                     FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD
                     GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA
                     KE"
     terminator      11810..12007
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
ORIGIN
        1 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat
       61 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
      121 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
      181 cacgacgttg taaaacgacg gccagtgttg taaaacgacg gccagtacca tttgagcatt
      241 ggaaacttcc tatcaaaaat tacatagaaa tttttgattt tggctatata aattacattt
      301 catgataaat gtacagctaa agtcactatg tccgagtggt taaggagtta gacttcaatc
      361 ctgtattcta gtcatctaat gggctttgcc cgcgcaggtt caaatcctgc tggtgacgga
      421 tggccggcat ggtcccagcc tcctcgctgg cgccggctgg gcaacacctt cgggtggcga
      481 atgggactag agaccgaaag tgaaacgtga tttcatgcgt cattttgaac attttgtaaa
      541 tcttatttaa taatgtgtgc ggcaattcac atttaattta tgaatgtttt cttaacatcg
      601 cggcaactca agaaacggca ggttcggatc ttagctacta gagaaagagg agaaatacta
      661 gatgcgtaaa ggcgaagagc tgttcactgg tgtcgtccct attctggtgg aactggatgg
      721 tgatgtcaac ggtcataagt tttccgtgcg tggcgagggt gaaggtgacg caactaatgg
      781 taaactgacg ctgaagttca tctgtactac tggtaaactg ccggttcctt ggccgactct
      841 ggtaacgacg ctgacttatg gtgttcagtg ctttgctcgt tatccggacc atatgaagca
      901 gcatgacttc ttcaagtccg ccatgccgga aggctatgtg caggaacgca cgatttcctt
      961 taaggatgac ggcacgtaca aaacgcgtgc ggaagtgaaa tttgaaggcg ataccctggt
     1021 aaaccgcatt gagctgaaag gcattgactt taaagaggac ggcaatatcc tgggccataa
     1081 gctggaatac aattttaaca gccacaatgt ttacatcacc gccgataaac aaaaaaatgg
     1141 cattaaagcg aattttaaaa ttcgccacaa cgtggaggat ggcagcgtgc agctggctga
     1201 tcactaccag caaaacactc caatcggtga tggtcctgtt ctgctgccag acaatcacta
     1261 tctgagcacg caaagcgttc tgtctaaaga tccgaacgag aaacgcgatc atatggttct
     1321 gctggagttc gtaaccgcag cgggcatcac gcatggtatg gatgaactgt acaaatgacc
     1381 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     1441 ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc
     1501 gtttataggt ctcagtttta gagctagaaa tagcaagtta aaataaggct agtccgttat
     1561 caacttgaaa aagtggcacc gagtcggtgc tttttcattt ttttttgttt tttatgtctg
     1621 tcatagctgt ttcctgtgtg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     1681 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     1741 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     1801 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     1861 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     1921 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     1981 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2041 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2101 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2161 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2221 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2281 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2341 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     2401 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     2461 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     2521 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2581 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     2641 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     2701 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     2761 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     2821 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     2881 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     2941 gagaaccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3001 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     3061 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     3121 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     3181 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     3241 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     3301 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     3361 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     3421 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     3481 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     3541 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     3601 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     3661 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     3721 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     3781 aagtgccacc tgacgcatgc cctacaacaa ctaagaaaat ggctatcatg cggaagctgg
     3841 tgagaagaac agcatcggga caagggaagg aagaacaaag acaaagaaaa caaaagaaag
     3901 caattgaaaa caaaacaaaa caattttcat tccttctctt atcattcctt ttcttttctt
     3961 ttctctcatt caacgcactc catcgtatcc gtattcctct tattttttct ctttctctat
     4021 atccatttct ttctctctag gtgtgtcctc tctctctctt caatttctct actccgcatt
     4081 ccaacgcatc cttcccccaa cctcccattt cctccttacg gcccgatagc gatcgtcttt
     4141 ccctcgctat cactcgctac cggcccctcc tctgcaccgt aacctcctac gtatttacca
     4201 tatcataaag ttttttccga cgcttatcgc tgaccccctg tcgccctcct attggcttcc
     4261 ggattatctt cttgtccata aggtgatcca tgcttcctga agattcccga aatgtgtcca
     4321 ctttggcggg gaatcattcc atccacttct ttctctctcg ctttcctcat tcggcgctcc
     4381 ccttccgcgt ctcattggtc ttccgctccg tttttgcttt gccgatgtta cttggggaga
     4441 ggtgcgataa tcctttcgca aaaactcggt ttgacgcctc ccatggtaaa tatagtgggt
     4501 ggtggacagg tgccttcgct tttctttaag caagagaatt gctgcaggtc gactctagag
     4561 atggataaga aatactctat tggccttgat atcggaacta acagtgtagg ttgggcagtc
     4621 ataacagatg aatacaaagt tccatctaag aaatttaaag tccttggaaa cactgaccga
     4681 catagtataa aaaagaattt aatcggcgca ttgcttttcg actctggtga aacggcagaa
     4741 gccacccgat tgaaacgaac agcaagacgt cgatatacgc gaagaaagaa ccgtatttgc
     4801 tatcttcagg agattttttc taatgaaatg gctaaggtcg acgactcctt tttccacaga
     4861 ttggaggaaa gtttccttgt cgaagaggac aaaaaacacg agcgtcaccc tatcttcgga
     4921 aacattgtag acgaagtggc atatcatgaa aagtatccta cgatatacca tttgcgtaaa
     4981 aagttggtcg actcaacgga caaggcagat ttgcgtttga tatacttggc tcttgcccac
     5041 atgattaaat ttcgtggcca tttcttaatt gaaggtgacc ttaacccaga taacagtgat
     5101 gtggacaagt tgttcattca attggttcag acctacaatc aattatttga ggaaaaccca
     5161 atcaatgcat ctggcgtaga tgcaaaggct attctttccg cacgcctttc aaaatcacgc
     5221 cgattagaga atcttatcgc tcagttacct ggcgagaaga aaaatggctt gttcggtaat
     5281 ttgatcgctt tgagtttggg ccttacacca aattttaagt ccaacttcga tcttgcagaa
     5341 gatgccaaat tacaattaag taaggatacg tacgacgatg atttggacaa tttacttgca
     5401 caaatcggtg accaatatgc cgatcttttc ttggcagcta aaaatttatc agacgcaatt
     5461 ttattgtccg atatccttcg tgtgaacacc gaaattacca aggccccact tagtgcttcc
     5521 atgataaaac gctatgatga gcatcatcag gacttaacat tacttaaagc cttggtgcgc
     5581 caacaattac ccgaaaagta caaagaaata ttcttcgacc agtccaagaa cggttatgcc
     5641 ggctacatcg atggtggtgc atcacaggag gagttctaca aattcattaa accaatctta
     5701 gaaaaaatgg atggtaccga agagttattg gtaaagttaa atcgcgaaga cttgcttcgc
     5761 aaacaacgaa cattcgacaa cggctccatc cctcatcaaa ttcaccttgg cgaattacat
     5821 gccatcttac gcagacaaga agacttttat ccattcttga aggataaccg cgagaaaatt
     5881 gagaagatcc ttacctttag aattccatac tacgtaggcc ctttggctag aggcaactcc
     5941 agatttgcat ggatgaccag aaaatctgag gaaaccatca cgccctggaa cttcgaagaa
     6001 gtcgtagata agggcgcaag tgcacagtcc tttattgaaa gaatgaccaa ttttgataag
     6061 aacttaccta acgaaaaggt cttaccaaag catagtttac tttatgaata cttcacggtg
     6121 tacaacgaat taacaaaagt caaatacgtt actgagggaa tgagaaagcc tgcattcttg
     6181 tccggagaac aaaagaaggc catagttgac cttcttttca aaacgaatcg aaaggtcacg
     6241 gtaaaacaac ttaaggagga ttactttaag aagatagaat gctttgatag tgttgagata
     6301 tcaggtgttg aagaccgatt caatgcatca ttgggaacgt accacgacct tttgaagatc
     6361 ataaaagata aagatttttt agacaatgag gagaacgaag acattttaga agacatcgtt
     6421 ttgacactta cgttatttga ggacagagag atgatcgagg agcgtcttaa aacgtatgca
     6481 catttattcg acgacaaagt gatgaaacaa cttaagcgca gaagatatac cggttgggga
     6541 agactttctc gaaagttgat caacggcatc cgcgataagc aaagtggaaa aaccattttg
     6601 gatttcttaa agtctgatgg ctttgctaat cgcaacttca tgcaattgat tcatgacgat
     6661 tcattaactt tcaaggaaga catacagaag gcccaagttt caggtcaggg agatagtttg
     6721 cacgagcaca tcgctaattt agctggttcc cctgctataa aaaagggtat acttcaaacc
     6781 gtaaaagtcg tagatgagtt ggtaaaagtc atgggcagac ataaaccaga aaatatagta
     6841 attgagatgg cccgagaaaa tcaaacgacg caaaagggcc aaaaaaactc tagagaaaga
     6901 atgaagcgta tcgaggaggg cataaaggag ttgggttctc agatattgaa ggagcacccc
     6961 gttgagaaca ctcagcttca gaatgagaag ctttatttat attatttaca aaacggtcgt
     7021 gatatgtatg tggatcagga gttggacata aacagattgt cagattacga tgtggatcat
     7081 atcgtccccc aatcattctt aaaggacgat tcaattgaca acaaagttct tacgcgtagt
     7141 gacaagaaca gaggcaaatc agacaatgta cctagtgagg aagtagtgaa aaagatgaag
     7201 aactattgga gacaacttct taatgctaag ttaataacgc agcgaaagtt tgataactta
     7261 actaaagccg aacgtggcgg actttcagag ttggataaag caggctttat caaacgtcaa
     7321 ttggtagaga ctcgtcagat cactaagcat gtggcccaaa tacttgattc acgcatgaat
     7381 actaagtacg atgaaaacga taaattgatt cgtgaggtta aggtaatcac gttgaaatct
     7441 aagcttgttt ccgactttcg taaggatttt caattctaca aggttcgaga gattaataat
     7501 taccaccatg cacatgacgc atacttgaac gcagtcgtag gtacagcatt gatcaagaag
     7561 tacccaaaat tagagtcaga atttgtctac ggtgactaca aagtatatga tgtccgaaag
     7621 atgatagcca agtctgagca agagatagga aaggccacag ctaagtattt tttttactcc
     7681 aacattatga atttctttaa gacagagatt actttagcta acggtgaaat acgaaaaaga
     7741 ccccttatcg aaacgaatgg agagacaggt gagattgttt gggacaaagg tcgagatttc
     7801 gccactgtga gaaaagtctt gagtatgcca caggtcaata ttgttaagaa aacagaagta
     7861 cagactggtg gattttctaa agaatccatt ttgcccaaga gaaattccga caaacttatt
     7921 gcacgcaaga aggattggga tcccaagaaa tacggcggtt tcgacagtcc cactgtagca
     7981 tactcagtgt tggtcgtggc taaggtggaa aagggtaagt ctaagaagct taaatctgtg
     8041 aaggaattgc ttggtatcac gatcatggag cgctcatctt ttgagaaaaa tcctatagac
     8101 tttcttgaag ctaaaggcta taaagaggtg aaaaaggacc ttataattaa gttaccaaag
     8161 tactctcttt tcgaacttga gaacggacgc aagagaatgc ttgccagtgc tggagaatta
     8221 cagaagggta atgagttagc attgccatca aagtacgtaa acttcttata tttagcctcc
     8281 cactatgaga aattaaaggg ttcacccgag gacaatgaac aaaagcaatt attcgtagaa
     8341 caacataaac actatttgga cgaaattata gagcaaatct ctgaattttc caaaagagtg
     8401 attttagctg acgcaaatct tgataaggtt ttatctgcct ataacaagca tcgcgacaaa
     8461 cccatcagag aacaagccga aaatatcatt cacttattca cacttacgaa tcttggagcc
     8521 cctgccgctt tcaagtattt tgatactacc atagaccgta aacgctatac gtctactaag
     8581 gaagtcttag acgccactct tattcaccaa agtataaccg gcttgtatga gacaagaatt
     8641 gatttgtctc aattgggcgg cgacagcagg gctgacccca agaagaagag gaaggtgtga
     8701 ttttggacct cgagtcatgt aattagttat gtcacgctta cattcacgcc ctccccccac
     8761 atccgctcta accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt
     8821 ttttatagtt atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct
     8881 gtacagacgc gtgtacgcat gtaacattat actgaaaacc ttgcttgaga aggttttggg
     8941 acgctcgaag gctttaattt gcgatccccg tctaagaaac cattattatc atgacattaa
     9001 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     9061 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     9121 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     9181 actatgcggc atcagagcag attgtactga gagtgcacca taccatcgat ggccaacctt
     9241 ccaattcatt aaatcgagtc ctccatcaga tgatttacaa ggttttgcat agaatcccta
     9301 agtaagagaa gttgttagtg atttcttcaa gtttactaaa atttacatac aaatggatga
     9361 tctcttctcc attgtttcct ttcttcttgc aaacgcgttt tacaaagaga tgacatgcta
     9421 ttttctgatt atttttttct atagttttct attttgtacg tccttgggag tccgaaatgt
     9481 aaaatcgggg tatggttagt agcggcagcg ttcattatgg tatataaaag ataaaaagta
     9541 ggggcaagcg agacagaatg ggatacaagg gcatcgtcta taattatagc taaaaattgt
     9601 attttaattt gtattttttg taattttatt tctcattgtt ttacttaaaa tgaatgcgaa
     9661 ttagagaaaa cttcaacgaa atgtcaaaat aagctcagca aaatatacaa ttttagggaa
     9721 agcgatcagc aacttattct ccgttgtaga ttctttttct tcataaattc caacaatgta
     9781 actgccaatt gatttttctc tagatgctaa taaattgaaa ctaatgaata taataacaca
     9841 ggttttgata aattgtaaac tcttactcaa aaaaagaatg ttaaaactaa tttaaccttt
     9901 ttataaaggc tcatggcgtt aaagaggata gtaaaaattc tacaattttt taatttaatg
     9961 tcacaattgt cttgcgaata taattctatt aattttcgct aactgctgtt ttagtactat
    10021 tactttcaat atgccttata tgcaacttta attcatgaag aatgcaatat ttttgcaatc
    10081 tttcttgtac tgagcgcttt ttatcattta gtactcatta tttaattttt cagtaaaggg
    10141 caataaataa atttgtagaa gatgcaatgt aatctcctct atccttttgc tcatatgttt
    10201 atgagtatac ctagtctaga aaggcttgta tttaaaatat tattcaataa aaattcacaa
    10261 tttttacgac atgtgctacc attcacttaa cttcctgatt ataaaattgg ttcgtttata
    10321 ctaattactt aagtaccttt aactaaacaa aatgcctata tatatattaa tttacaatga
    10381 gtgtcagata agtcactatg tccgagtggt taaggagtta gactcgccat cgaaatggag
    10441 gcccagaata ccctccttga cagtcttgac gtgcgcagct caggggcatg atgtgactgt
    10501 cgcccgtaca tttagcccat acatccccat gtataatcat ttgcatccat acattttgat
    10561 ggccgcacgg cgcgaagcaa aaattacggc tcctcgctgc ggacctgcga gcagggaaac
    10621 gctcccctca cagacgcgtt gaattgtccc cacgccgcgc ccctgtagag aaatataaaa
    10681 ggttaggatt tgccactgag gttcttcttt catatacttc cttttaaaat cttgctagga
    10741 tacagttctc acatcacatc cgaacataaa caaccatggg taaaaagcct gaactcaccg
    10801 cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcggc
    10861 tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc
    10921 tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg
    10981 catcggccgc gctcccgatt ccggaagtgc ttgacattgg ggaattcagc gagagcctga
    11041 cctattgcat ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac
    11101 tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg gccgatctta
    11161 gccagacgag cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc
    11221 gtgatttcat atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg
    11281 acaccgtcgg tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact
    11341 gccccgaagt ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca
    11401 atggccgcat aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg
    11461 aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct
    11521 acttcgagcg gaggcatccg gagcttgcag gatcgccgcg gctccgggcg tatatgctcc
    11581 gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt
    11641 gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac
    11701 aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg tgtagaagta ctcgccgata
    11761 gtggaaaccg acgccccagc actcgtccga gggcaaagga ataatcagta ctgacaataa
    11821 aaagattctt gttttcaaga acttgtcatt tgtatagttt ttttatattg tagttgttct
    11881 attttaatca aatgttagcg tgatttatat tttttttcgc ctcgacatca tctgcccaga
    11941 tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc
    12001 tatactg
//
