LOCUS       sequence-181276-       13333 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPRi vector for Yarrowia lipolytica, repressing KU70 and KU80 for
            enhanced HR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13333)
  AUTHORS   Schwartz C, Frogue K, Ramesh A, Misa J, Wheeldon I
  TITLE     CRISPRi repression of nonhomologous end-joining for enhanced genome 
            engineering via homologous recombination in Yarrowia lipolytica.
  JOURNAL   Biotechnol Bioeng. 2017 Aug 19. doi: 10.1002/bit.26404.
  PUBMED    28832943
REFERENCE   2  (bases 1 to 13333)
  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: "Biotechnol 
            Bioeng. 2017 Aug 19. doi: 10.1002/bit.26404."
COMMENT     Sequence Label: pCRISPRi_Mxi1_yl_NHEJ
FEATURES             Location/Qualifiers
     source          1..13333
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_RNA        1112..1187
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        1798..1873
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             join(3078..3104,3105..3107,3108..5594,5595..5597,
                     5598..7181)
                     /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 H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     CDS             7194..7214
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      7396..7643
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(7412..7430)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(7662..7678)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7662..7678)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7675..7697)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7686..7702
                     /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(7710..7716,7717..7734,7735..7740))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7755..7776
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7893..7910)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8064..8652)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8144..8163)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8823..9614,9615..9683))
                     /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     9446..9465
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9684..9788)
                     /gene="bla"
                     /label=AmpR promoter
     misc_RNA        10282..10357
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acccgctttt
      421 cgtagataat ggaatacaaa tggatatcca gagtatacac atggatagta tacactgaca
      481 cgacaattct gtatctcttt atgttaacta ctgtgaggcg ttaaatagag cttgatatat
      541 aaaatgttac atttcacagt ctgaactttt gcagattacc taatttggta agatattaat
      601 tatgaactga aagttgatgg catccctaaa tttgatgaaa gggggatccc ccagttgcaa
      661 aagttgacac aactctagat ctgcttccaa atatagaatc ataacaaggg ttagggtgtg
      721 attatataat attggtctta attgatgtgc tagggcttta aaagttggtt aaaataacgc
      781 tctaatgcct ttttaatata ttgtcttttt caaaatctca aatcggacac ttcttcgtgt
      841 atgagactcc attttttggc tccgtcacgt gatatgtatt atcagctata gtggtgtaaa
      901 caaagttttt tactagctgt aatggcattt tgtcggagtg gtaaatcgcc ttcttgttgt
      961 gcgttcgagt tctggactct gcactgggct actttgaaaa atacctctaa tgcgccgatg
     1021 gtttagtggt aaaatccatc gttgccatcg atgggccccc ggttcgattc cgggtcggcg
     1081 caggttgacg tctggtatgg cagtgtgctc tgttttagag ctagaaatag caagttaaaa
     1141 taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt tttttacgtc
     1201 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccaga
     1261 tcctctagag tcgaagcggc cgctatgtct gataaaagga tgtaactact tgatgtcttg
     1321 tgacgttctt agagatccag ttgcaaaagt tgacacaact ctagatctgc ttccaaatat
     1381 agaatcataa caagggttag ggtgtgatta tataatattg gtcttaattg atgtgctagg
     1441 gctttaaaag ttggttaaaa taacgctcta atgccttttt aatatattgt ctttttcaaa
     1501 atctcaaatc ggacacttct tcgtgtatga gactccattt tttggctccg tcacgtgata
     1561 tgtattatca gctatagtgg tgtaaacaaa gttttttact agctgtaatg gcattttgtc
     1621 ggagtggtaa atcgccttct tgttgtgcgt tcgagttctg gactctgcac tgggctactt
     1681 tgaaaaatac ctctaatgcg ccgatggttt agtggtaaaa tccatcgttg ccatcgatgg
     1741 gcccccggtt cgattccggg tcggcgcagg ttgacgtggt tgtgtgtgta tgctgttgtt
     1801 ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg aaaaagtggc
     1861 accgagtcgg tgcttttttt tacgtctaag aaaccattat tatcatgaca ttaacctata
     1921 aaaataggcg tatcacgagg cccagatcct ctagagtcga agcggccgct atgtctgata
     1981 aaaggatgta acgttcgaag gtaccaagga agcatgcggt acccgaattc ctgaggtgtc
     2041 tcacaagtgc cgtgcagtcc cgcccccact tgcttctctt tgtgtgtagt gtacgtacat
     2101 tatcgagacc gttgttcccg cccacctcga tccggtctag actgaggtgt ctcacaagtg
     2161 ccgtgcagtc ccgcccccac ttgcttctct ttgtgtgtag tgtacgtaca ttatcgagac
     2221 cgttgttccc gcccacctcg atccggggat ccctgaggtg tctcacaagt gccgtgcagt
     2281 cccgccccca cttgcttctc tttgtgtgta gtgtacgtac attatcgaga ccgttgttcc
     2341 cgcccacctc gatccgggtc gacctgaggt gtctcacaag tgccgtgcag tcccgccccc
     2401 acttgcttct ctttgtgtgt agtgtacgta cattatcgag accgttgttc ccgcccacct
     2461 cgatccggga gctcctgagg tgtctcacaa gtgccgtgca gtcccgcccc cacttgcttc
     2521 tctttgtgtg tagtgtacgt acattatcga gaccgttgtt cccgcccacc tcgatccggt
     2581 ctagactgag gtgtctcaca agtgccgtgc agtcccgccc ccacttgctt ctctttgtgt
     2641 gtagtgtacg tacattatcg agaccgttgt tcccgcccac ctcgatccgg ggatccctga
     2701 ggtgtctcac aagtgccgtg cagtcccgcc cccacttgct tctctttgtg tgtagtgtac
     2761 gtacattatc gagaccgttg ttcccgccca cctcgatccg ggtcgacctg aggtgtctca
     2821 caagtgccgt gcagtcccgc ccccacttgc ttctctttgt gtgtagtgta cgtacattat
     2881 cgagaccgtt gttcccgccc acctcgatcc gggcatgcct gcagaagctt ttgtggttgg
     2941 gactttagcc aagggtataa aagaccaccg tccccgaatt acctttcctc ttcttttctc
     3001 tctctccttg tcaactcaca cccgaaatcg ttaagcattt ccttctgagt ataagaatca
     3061 ttcaaaggcg cgccacaatg gataagaaat actccattgg cctggccatc ggaaccaact
     3121 ccgtgggttg ggccgtgatc accgatgagt acaaggtgcc ctctaagaaa ttcaaggtcc
     3181 tgggcaacac cgaccgacac tccatcaaga agaacctgat cggcgctctg ctcttcgact
     3241 ctggcgagac cgctgaggcc acccgactga agcgaaccgc tcgaagacga tacacccgaa
     3301 gaaagaaccg aatctgttac ctgcaggaga tcttctctaa cgagatggcc aaggtggacg
     3361 actctttctt ccaccgactg gaggagtctt tcctggtgga ggaggacaag aagcacgagc
     3421 gacaccccat cttcggcaac atcgtggacg aggtggccta ccacgagaag taccccacca
     3481 tctaccacct gcgaaagaag ctggtggact ctaccgacaa ggccgacctg cgactgatct
     3541 acctggccct ggcccacatg atcaagttcc gaggccactt cctgatcgag ggcgacctga
     3601 accccgacaa ctctgacgtg gacaagctgt tcatccagct ggtgcagacc tacaaccagc
     3661 tcttcgaaga gaaccccatt aacgcttctg gcgtggatgc taaggccatc ctgtctgccc
     3721 gactgtctaa gtctcgacga ctcgagaacc tgattgctca gctccccgga gagaagaaga
     3781 acggtctgtt cggaaacctg attgctctgt ccctgggtct cacccctaac ttcaagtcca
     3841 acttcgatct ggctgaggac gctaagctgc agctgtctaa ggacacctac gacgatgacc
     3901 tggataacct gctcgcccag attggcgacc agtacgccga cctgttcctg gccgccaaga
     3961 acctgtctga cgccatcctg ctgtctgaca tcctgcgagt gaacaccgag atcaccaagg
     4021 cccccctgtc tgcctccatg attaagcgat acgatgagca ccaccaggat ctgaccctcc
     4081 tcaaggctct ggtccgacag cagctgcccg agaagtacaa ggagattttc ttcgaccagt
     4141 ctaagaacgg ctacgccggc tacatcgacg gcggcgcctc tcaggaggag ttctacaagt
     4201 tcattaagcc catcctggag aagatggacg gaaccgagga actgctcgtg aagctgaacc
     4261 gagaggacct cctgcgaaag cagcgaacct tcgacaacgg ctctatcccc caccagatcc
     4321 acctgggcga gctgcacgcc atcctgcgac gacaggagga cttctacccc ttcctgaagg
     4381 acaaccgaga gaagatcgag aagatcctga ccttccgaat cccctactac gtgggacccc
     4441 tggcccgagg aaactctcga ttcgcttgga tgacccgaaa gtctgaggag accattaccc
     4501 cctggaactt cgaggaggtg gtggataagg gcgcctctgc tcagtctttc atcgagcgaa
     4561 tgaccaactt cgacaagaac ctccccaacg agaaggtcct gcccaagcac tctctgctct
     4621 acgagtactt caccgtctac aacgagctca ccaaggtcaa gtacgtgacc gagggaatgc
     4681 gaaagcccgc tttcctgtct ggagagcaga agaaggctat tgtggatctg ctcttcaaga
     4741 ctaaccgaaa ggtcaccgtc aagcagctga aggaggatta cttcaagaag attgagtgtt
     4801 tcgattctgt cgagatctcc ggcgtcgagg accgattcaa cgcctctctg ggtacctacc
     4861 acgacctgct gaagattatc aaggacaagg atttcctgga taacgaggag aacgaggata
     4921 ttctcgagga cattgtcctg accctcaccc tgttcgagga tcgagagatg attgaggagc
     4981 gactcaagac ctacgctcac ctgttcgacg acaaggtgat gaagcagctg aagcgacgac
     5041 gatacaccgg ctggggccga ctgtctcgaa agctgatcaa cggcatccga gacaagcagt
     5101 ctggcaagac catcctggac ttcctgaagt ctgacggctt cgccaaccga aacttcatgc
     5161 agctgatcca cgacgactct ctgaccttca aggaggacat ccagaaggcc caggtgtctg
     5221 gccagggcga ctctctgcac gagcacatcg ccaacctggc cggctctccc gccattaaga
     5281 aaggtatcct gcagaccgtc aaggtggtcg atgagctcgt caaggtgatg ggccgacaca
     5341 agcccgagaa cattgtcatt gagatggctc gagagaacca gactactcag aagggtcaga
     5401 aaaactcccg agagcgaatg aagcgaattg aggaaggtat taaggagctg ggatcccaga
     5461 ttctcaagga gcatcccgtg gagaacactc agctccagaa cgagaagctg tacctgtact
     5521 atctgcagaa cggtcgagac atgtacgtcg accaggagct ggatatcaac cgactctccg
     5581 actacgatgt ggacgccatt gtgccccagt ccttcctgaa ggacgattct atcgataaca
     5641 aggtgctgac ccgatccgac aagaaccgag gcaagtctga caacgtgccc tccgaggagg
     5701 tggtcaagaa gatgaagaac tactggcgac agctgctgaa cgccaagctg attacccagc
     5761 gaaagttcga caacctgacc aaggccgagc gaggcggcct gtctgagctg gacaaggccg
     5821 gcttcatcaa gcgacagctg gtggagaccc gacagatcac caagcacgtg gcccagatcc
     5881 tggactctcg aatgaacacc aagtacgacg agaacgacaa gctgatccga gaggtgaagg
     5941 tgatcaccct gaagtctaag ctggtgtctg acttccgaaa ggacttccag ttctacaagg
     6001 tgcgagagat taacaactac caccacgccc acgatgccta cctgaacgct gtcgtgggca
     6061 ccgccctcat caagaagtat cccaagctgg agtccgagtt cgtctacggc gactacaagg
     6121 tctacgatgt gcgaaaaatg attgccaagt ccgagcagga gattggcaag gctaccgcca
     6181 agtacttctt ctactccaac attatgaact tcttcaagac cgagattacc ctggctaacg
     6241 gcgagattcg aaagcgaccc ctcattgaga ccaacggaga gaccggtgag atcgtgtggg
     6301 acaagggacg agacttcgcc accgtgcgaa aggtgctgtc tatgccccag gtgaacatcg
     6361 tgaagaagac cgaggtgcag accggaggtt tctctaagga gtccatcctg cccaagcgaa
     6421 actctgacaa gctgatcgcc cgaaagaagg actgggaccc caagaagtac ggaggtttcg
     6481 actctcccac cgtggcttac tctgtgctgg tggtggccaa ggtggagaag ggcaagtcta
     6541 agaagctgaa gtctgtgaag gagctgctgg gcattaccat catggagcga tcttctttcg
     6601 agaagaaccc cattgacttc ctggaggcca agggatacaa ggaggtgaag aaagatctga
     6661 ttatcaagct ccccaagtac tctctgttcg agctggagaa cggacgaaag cgaatgctgg
     6721 cctctgccgg cgagctgcag aagggaaacg agctggccct gccctccaag tacgtcaact
     6781 tcctgtacct cgcctcccat tacgagaagc tgaagggctc tcccgaggat aacgagcaga
     6841 agcagctctt cgtggagcag cataagcact acctggacga gatcatcgag cagatctctg
     6901 agttctctaa gcgagtgatc ctggctgacg ccaacctgga taaggtgctg tctgcttaca
     6961 acaagcaccg agacaagccc attcgagagc aggctgagaa catcattcac ctgttcaccc
     7021 tgaccaacct gggagccccc gctgccttca agtacttcga caccaccatc gaccgaaagc
     7081 gatacacctc taccaaggag gtgctggacg ccaccctgat ccaccagtct atcaccggcc
     7141 tgtacgagac ccgaatcgac ctgtctcagc tgggcggcga ctctcgagcc gaccccaaga
     7201 agaagcgaaa ggtgggatct ggtggaggct cctctaagct gggcggctct ggcggctcta
     7261 tggagcgagt gcgaatgatc aacgtgcagc gactgctgga ggccgccgag ttcctggagc
     7321 gacgagagcg agagtgcgag cacggctacg cctcttcttt cccctctatg ccctctcccc
     7381 gaggctaggc tagcctcatg taattagtta tgtcacgctt acattcacgc cctccctcca
     7441 catccgctct aaccgaaaag gaaggagtta gacaacctga agtctaggtc cctatttatt
     7501 tttttatagt tatgttagta ttaagaacgt tatttatatt tcaaattttt cttttttttc
     7561 tgtacagacg cgtgtacgca tgtaacatta tactgaaaac cttgcttgag aaggttttgg
     7621 gacgctcgaa ggctttaatt tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg
     7681 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     7741 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     7801 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     7861 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     7921 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     7981 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     8041 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     8101 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     8161 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     8221 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     8281 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     8341 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     8401 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     8461 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     8521 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     8581 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     8641 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     8701 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     8761 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     8821 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     8881 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     8941 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     9001 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     9061 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     9121 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     9181 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     9241 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     9301 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     9361 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     9421 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     9481 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     9541 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     9601 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     9661 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     9721 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     9781 gttccgcgca catttccccg aaaagtgcca cctgacgtcc ccagttgcaa aagttgacac
     9841 aactctagat ctgcttccaa atatagaatc ataacaaggg ttagggtgtg attatataat
     9901 attggtctta attgatgtgc tagggcttta aaagttggtt aaaataacgc tctaatgcct
     9961 ttttaatata ttgtcttttt caaaatctca aatcggacac ttcttcgtgt atgagactcc
    10021 attttttggc tccgtcacgt gatatgtatt atcagctata gtggtgtaaa caaagttttt
    10081 tactagctgt aatggcattt tgtcggagtg gtaaatcgcc ttcttgttgt gcgttcgagt
    10141 tctggactct gcactgggct actttgaaaa atacctctaa tgcgccgatg gtttagtggt
    10201 aaaatccatc gttgccatcg atgggccccc ggttcgattc cgggtcggcg caggttgacg
    10261 tcgatgtata gttataattg ggttttagag ctagaaatag caagttaaaa taaggctagt
    10321 ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt tttttacgtc taagaaacca
    10381 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccaga tcctctagag
    10441 tcgaagcggc cgctatgtct gataaaagga tgtaacatag gcaagctgct cgtgagtgtt
    10501 gagtacgaac cttagatcca aatcacccgc acccacggat atacttgctt gaatatacag
    10561 tagtatgcgg ccgcttcgac accatatcat ataaaactaa caatgcattg aattaatacg
    10621 aagactaccc gttgctatct ccacaccgtt atctccacgg tccaaaggct gctcaatgtg
    10681 ctgcatacgt aacgtggggt gcaaccttga gcacatagta ctttccgaaa accggcgata
    10741 attaagtgtg cactccaact tttcacactg agcataaaat gtggagaaga aatcacacta
    10801 aaaagtcagg tagactggaa aaatgcgcca tgaaataata tctcttgcta cagtaatgcc
    10861 cagcatcgag gggtattgtg tcaccaacac tatagtggca gctgaagcgc tcgtgattgt
    10921 agtatgagtc tttattggtg atgggaagag tccactcaat attctcgtta ctgccaaaac
    10981 accacggtaa tcggccagac accatggatg tagatcacca agcctgtgaa tgttattcga
    11041 gctaaaatgc acatggttgg tgaaaggagt agttgctgtc gaattccgtc gtcgcctgag
    11101 tcatcattta tttaccagtt ggccacaaac ccttgacgat ctcgtatgtc ccctccgaca
    11161 tactcccggc cggctggggt acgttcgata gcgctatcgg catcgacaag gtttgggtcc
    11221 ctagccgata ccgcactacc tgagtcacaa tcttcggagg tttagtcttc cacatagcac
    11281 gggcaaaagt gcgtatatat acaagagcgt ttgccagcca cagattttca ctccacacac
    11341 cacatcacac atacaaccac acacatccac aatggaaccc gaaactaaga agaccaagac
    11401 tgactccaag aagattgttc ttctcggcgg cgacttctgt ggccccgagg tgattgccga
    11461 ggccgtcaag gtgctcaagt ctgttgctga ggcctccggc accgagtttg tgtttgagga
    11521 ccgactcatt ggaggagctg ccattgagaa ggagggcgag cccatcaccg acgctactct
    11581 cgacatctgc cgaaaggctg actctattat gctcggtgct gtcggaggcg ctgccaacac
    11641 cgtatggacc actcccgacg gacgaaccga cgtgcgaccc gagcagggtc tcctcaagct
    11701 gcgaaaggac ctgaacctgt acgccaacct gcgaccctgc cagctgctgt cgcccaagct
    11761 cgccgatctc tcccccatcc gaaacgttga gggcaccgac ttcatcattg tccgagagct
    11821 cgtcggaggt atctactttg gagagcgaaa ggaggatgac ggatctggcg tcgcttccga
    11881 caccgagacc tactccgttc ctgaggttga gcgaattgcc cgaatggccg ccttcctggc
    11941 ccttcagcac aacccccctc ttcccgtgtg gtctcttgac aaggccaacg tgctggcctc
    12001 ctctcgactt tggcgaaaga ctgtcactcg agtcctcaag gacgaattcc cccagctcga
    12061 gctcaaccac cagctgatcg actcggccgc catgatcctc atcaagcagc cctccaagat
    12121 gaatggtatc atcatcacca ccaacatgtt tggcgatatc atctccgacg aggcctccgt
    12181 catccccggt tctctgggtc tgctgccctc cgcctctctg gcttctctgc ccgacaccaa
    12241 cgaggcgttc ggtctgtacg agccctgtca cggatctgcc cccgatctcg gcaagcagaa
    12301 ggtcaacccc attgccacca ttctgtctgc cgccatgatg ctcaagttct ctcttaacat
    12361 gaagcccgcc ggtgacgctg ttgaggctgc cgtcaaggag tccgtcgagg ctggtatcac
    12421 taccgccgat atcggaggct cttcctccac ctccgaggtc ggagactttg ttgccaacaa
    12481 ggtcaaggag ctgctcaaga aggagtaagt cgtttctacg acgcattgat ggaaggagca
    12541 aactgacgcg cctgcgggtt ggtctaccgg cagggtccgc tagtgtataa gactctataa
    12601 aaagggccca gccctgctaa tgaaatgatg atttataatt taccggtgta gcaaccttga
    12661 ctagaagaag cagattgggt gtgtttgtag tggaggacag tggtacgttt tggaaacagt
    12721 cttcttgaaa gtgtcttgtc tacagtatat tcactcataa cctcaatagc caagggtgta
    12781 gtcggtttat taaaggaagg gagttgtggc tgatgtggat agatatcttt aaagctggcg
    12841 actgcaccca acgagtgtgg tggtagcttg ttactgtata ttcggtaaga tatattttgt
    12901 ggggttttta gtggtgtttg gtaggttagt gtctggtata tgagttgtag gcatgacaat
    12961 ttggaaaggg gtggactttg ggaatattgt gggatttcaa taccttagtt tgtacagggt
    13021 aattgttaca aatgatacaa agaactgtat ttcttttcat ttgttttaat tggttgtata
    13081 tcaagtccgt tagacgagct cagtgccatg gcttttggca ctgtatttca tttttagagg
    13141 tacactacat ccagtgaggt atggtaaggt tgagggcata atgaaggcac cttgtactga
    13201 cagtcacaga cctctcaccg agaattttat gagatatact cgggttcatt ttaggctccg
    13261 attcgattca aattattact gtcgaaatcg gttgagcatc cgttgatttc cgaacagatc
    13321 tggccctttc gtc
//
