LOCUS       sequence-496316-       11079 bp    DNA     circular SYN 07-JAN-2026
DEFINITION  Expresses nucleus-localized eNme2.C.NR-nCas9 (D16A) fused to 
            PolI5M-Delta and mCherry using a CMV promoter. Includes a GFP 
            cassette flanked by BsmbI cut sites to knock in and coexpress 
            user-defined gRNAs..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11079)
  AUTHORS   Hurtado JE, Schieferecke AJ, Halperin SO, Guan J, Aidlen D, Schaffer
            DV, Dueber JE
  TITLE     Nickase fidelity drives EvolvR-mediated diversification in mammalian
            cells.
  JOURNAL   Nat Commun. 2025 Apr 19;16(1):3723. doi: 10.1038/s41467-025-58414-0.
  PUBMED    40253348
REFERENCE   2  (bases 1 to 11079)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 7, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2025-04-19"; volume: "16(1):3723. doi"; pages: " 
            10.1038/s41467-025-58414-0"
COMMENT     Sequence Label: pEvolvR-mCherry-eNme2.C.NR-nCas9(D16A)-PolI5M-Delta
FEATURES             Location/Qualifiers
     source          1..11079
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(913..918,919..936,937..943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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)."
     primer_bind     956..978
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     975..991
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1153..1401
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1153..1173
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1324..1343
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     RBS             1557..1568
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1575..2288
                     /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      2300..2371
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2387..2414
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_RNA        2456..2548
                     /label=Nm tracrRNA
                     /note="trans-activating CRISPR RNA for the Neisseria 
                     meningitidis CRISPR/Cas9 system (Hou et al., 2013)"
     enhancer        2578..2957
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2958..3161
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3112..3132
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3180..3189
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3192..3212
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3237..3941
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(3379..3397)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3647..3666
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             7200..7226
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             7206..7232
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             7245..7265
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(9279..9298)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9280..9298)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9305..9322)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9305..9321)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9314..9336)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9463..9918
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9550..9569)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9760..9781
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        9944..10048
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10049..10117,10118..10909)
                     /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     complement(10267..10286)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagctcg aaattaaccc
     1021 tcactaaagg gaacaaaagc tggagctcca ccgcggtggc ggccgctctt aaggggtgca
     1081 gcggcctccg cgccgggttt tggcgcctcg gtgagtgagt gtgtgcgtgt tggatccaag
     1141 gtcgggcagg aagagggcct atttcccatg attccttcat atttgcatat acgatacaag
     1201 gctgttagag agataattag aattaatttg actgtaaaca caaagatatt agtacaaaat
     1261 acgtgacgta gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa
     1321 atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc
     1381 ttgtggaaag gacgaaacac ctgagacgga aagtgaaacg tgatttcatg cgtcattttg
     1441 aacattttgt aaatcttatt taataatgtg tgcggcaatt cacatttaat ttatgaatgt
     1501 tttcttaaca tcgcggcaac tcaagaaacg gcaggttcgg atcttagcta ctagagaaag
     1561 aggagaaata ctagatgcgt aaaggcgaag agctgttcac tggtgtcgtc cctattctgg
     1621 tggaactgga tggtgatgtc aacggtcata agttttccgt gcgtggcgag ggtgaaggtg
     1681 acgcaactaa tggtaaactg acgctgaagt tcatctgtac tactggtaaa ctgccggttc
     1741 cttggccgac tctggtaacg acgctgactt atggtgttca gtgctttgct cgttatccgg
     1801 accatatgaa gcagcatgac ttcttcaagt ccgccatgcc ggaaggctat gtgcaggaac
     1861 gcacgatttc ctttaaggat gacggcacgt acaaaacgcg tgcggaagtg aaatttgaag
     1921 gcgataccct ggtaaaccgc attgagctga aaggcattga ctttaaagag gacggcaata
     1981 tcctgggcca taagctggaa tacaatttta acagccacaa tgtttacatc accgccgata
     2041 aacaaaaaaa tggcattaaa gcgaatttta aaattcgcca caacgtggag gatggcagcg
     2101 tgcagctggc tgatcactac cagcaaaaca ctccaatcgg tgatggtcct gttctgctgc
     2161 cagacaatca ctatctgagc acgcaaagcg ttctgtctaa agatccgaac gagaaacgcg
     2221 atcatatggt tctgctggag ttcgtaaccg cagcgggcat cacgcatggt atggatgaac
     2281 tgtacaaatg accaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     2341 ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg
     2401 tgggcctttc tgcgtttata cgtctcagtt gtagctccct ttctcatttc ggaaacgaaa
     2461 tgagaaccgt tgctacaata aggccgtctg aaaagatgtg ccgcaacgct ctgcccctta
     2521 aagcttctgc tttaaggggc atcgtttatt ttccgcggcc tctagactcg aggcgttgac
     2581 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
     2641 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     2701 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     2761 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     2821 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     2881 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     2941 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     3001 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     3061 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     3121 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta actaccggtg
     3181 ccaccatggc cccaaagaag aagcggaagg tcggtatcca cggagtccca gcagccgtga
     3241 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca
     3301 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct
     3361 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct
     3421 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg
     3481 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga
     3541 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt
     3601 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga
     3661 agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga
     3721 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca
     3781 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca
     3841 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg
     3901 agggccgcca ctccaccggc ggcatggacg agctgtacaa gggatccggt atggccgcct
     3961 tcaagcctaa cccaatcaat tacatcctgg gactggcaat cggaatcgca tccgtgggat
     4021 gggctatggt ggagatcgac gaggaggaga atcctatccg gctgatcgat ctgggcgtga
     4081 gagtgtttga gagggccgag gtgccaaaga ccggcgattc tctggctatg gcccggagac
     4141 tggcacggag cgtgaggcgc ctgacacgga gaagggcaca caggctgctg agggcacgcc
     4201 ggctgctgaa gagagagggc gtgctgcagg cagcagactt cgatgagaat ggcctgatca
     4261 caagcctgcc aaacaccccc tggcagctga gagcagccgc cctggacagg aagctgacac
     4321 cactggagtg gtctgccgtg ctgctgcacc tgatcaagca ccgcggctac ctgagccagc
     4381 ggaagaacga gggagagaca gcagccaagg agctgggcgc cctgctgaag ggagtggcca
     4441 acaatgccca cgccctgcag accggcgatt tcaggacacc tgccgagctg gccctgaata
     4501 agtttgagaa ggagtccggc cacatcagaa accagagggg cgactatagc cacaccttct
     4561 cccgcaagga tctgcaggcc gagctgatcc tgctgttcga gaagcagaag gagtttggca
     4621 atccacacgt gagcggaggc ctgaaggagg gaatcgaaac cctgctgatg acacagaggc
     4681 ctgccctgtc cggcgacgca gtgcagaaga tgctgggaca ctgcaccctg gagcctaccg
     4741 agccaaaggc cgccaagaac acctacacag ccgagcggtt tatctggctg acaaagctga
     4801 acaatctgag aatcctggag cagggatccg agaggccact gaccgacaca gagaggtcca
     4861 ccctgatgga tgagccttac cggaagtcta agctgacata tgcccaggcc agaaagctgc
     4921 tgggcctgga ggacaccgcc ttctttaagg gcctgagata cggcaaggat aatgccgagg
     4981 cctccacact gatggagatg aaggcctatc acgccatctc tcgcgccctg gagaaggagg
     5041 gcctgaagga caagaagtcc cccctgaacc tgagctccga gctgcaggat gagatcggca
     5101 ccgccttctc tctgtttaag accgacgagg atatcacagg ccgcctgaag gacagggtgc
     5161 agcctgagat cctggaggcc ctgctgaagc acatctcttt cgataagttt gtgcagatca
     5221 gcctgaaggc cctgagaagg atcgtgccac tgatggagca gggcaagcgg tacgacgagg
     5281 cctgcgccga gatctacggc gtgcactatg gcaagaagaa cacagaggag aagatctatc
     5341 tgccccctat ccctgccgac gagatcagaa atcctgtggt gctgagggcc ctgtcccagg
     5401 caagaaaagt gatcaacgga gtggtgcgcc ggtacggatc tccagcccgg atccacatcg
     5461 aaaccgccag agaagtgggc aagagcttca aggaccggaa ggagatcgag aagagacagg
     5521 aggagaatcg caaggatcgg gagaaggccg ccgccaagtt tagggagtac ttccctaact
     5581 ttgtgggcga gccaaagtct aaggacatcc tgaagctgcg cctgtacgag cagcagcacg
     5641 gcaagtgtct gtatagcggc aaggagatca atctggtgcg gctgaacgag aagggctatg
     5701 tggagatcga tcacgccctg cctttctcca gaacctggga cgattctttt aacaataagg
     5761 tgctggtgct gggcagcgag aaccagaata agggcaatca gacaccatac gagtatttca
     5821 atggcaagga caactccagg gagtggcagg agttcaaggc ccgcgtggaa acctctagat
     5881 ttcccaggag caagaagcag cggatcctgc tgcagaagtt cgacgaggat ggctttaagg
     5941 agtgcaacct gaatgacacc agatacgtga accggttcct gtgccagttt gtggccgatc
     6001 acatcctgct gaccggcaag ggcaagagaa gggtgttcgc ctctaatggc cagatcacaa
     6061 acctgctgag gggattttgg ggactgagga aggtgcgggc agagaatgac agacaccacg
     6121 cactggatgc agtggtggtg gcatgcagca ccgtggcaat gcagcagaag atcacaagat
     6181 tcgtgaggta taaggagatg aacgcctttg acggcaagac catcgataag gagacaggca
     6241 aggtgctgca ccagaagacc cacttccccc agccttggga gttctttgcc caggaagtga
     6301 tgatccgggt gttcggcaag ccagacggca agcctgagtt tgaggaggcc gataccccag
     6361 agaagctgag gacactgctg gcagagaagc tgtctagcag gccagaggca gtgcacgagt
     6421 acgtgacccc actgttcgtg tccagggcac ccaatcggaa gatgtctggc gcccacaagg
     6481 acacactgag aagcgccaag aggtttgtga agcacaacga gaagatctcc gtgaagagag
     6541 tgtggctgac cgagatcaag ctggccgatc tggagaacat ggtgaattac aagaacggca
     6601 gggagatcga gctgtatgag gccctgaagg caaggctgga ggcctacgga ggaaatgcca
     6661 agcaggcctt cgacccaaag gataacccct tttataagaa gggaggacag ctggtgaagg
     6721 ccgtgcgggt ggagaagacc cagaagagcg gcgtgctgct gaataagaag aacgcctaca
     6781 caatcgccga caatggcgat atggtgagag tggacgtgtt ctgtaaggtg gataagaagg
     6841 gcaagaatca gtactttatc gtgcctatct atgcctggca ggtggccgag aacatcctgc
     6901 cagacatcga ttgcaagggc tacagaatcg acgatagcta tacattctgt ttttccctgc
     6961 acaagtatga cctgatcgcc ttccagaagg atgagaagtc caaggtggag tttgcctact
     7021 atatcaattg cgactcctct agcggcggct tctacctggc ctggcacgat aagggcagcc
     7081 gcgagcagcg ctttcgcatc tccacccaga atctggccct gatccagaag tatcaggtga
     7141 acgagctggg caaggagatc aggccatgtc ggctgaagaa gcgcccaccc gtgcgggcct
     7201 atccctatga cgtgcccgat tatgccagcc tgggcagcgg ctcccccaag aaaaaacgca
     7261 aggtggaaga tcctaagaaa aagcggaaag gttctagtga aaccccggga acaagtgagt
     7321 cggccacccc tgaaggtgga tcagggggta gcggatcctc ttacgataat tatgttacga
     7381 ttctggacga ggaaacctta aaggcttgga tcgctaaatt agagaaggct cctgttttcg
     7441 ctttcgacac ggaaacggat tctctggaca atattagtgc gaatcttgtt ggtctgagtt
     7501 tcgcaattga accgggtgtt gctgcttaca tccctgtggc acacgactac ctggacgctc
     7561 cggaccagat ttcacgtgaa cgcgctctgg aactgctgaa gcctttatta gaggacgaga
     7621 aagctttgaa agttggtcag aatttgaagt atgctcgtgg aatcttagct aattatggta
     7681 tcgagttgcg cggtatcgct ttcgacacga tgttggaatc ttatatcctg aactctgtcg
     7741 ctggtcgcca tgacatggac tctctggctg agcgctggct gaaacataag acgattacct
     7801 tcgaggaaat cgcaggaaag ggtaagaacc agctcacgtt caatcaaatc gctctggagg
     7861 aagctggtcg ctatgctgct gaggacgctg acgttactct gcaactgcac ttgaagatgt
     7921 ggcctgactt gcagaagcat aagggtccac tgaatgtttt tgaaaacatt gagatgcctt
     7981 tggttccagt tctgtctcgt atcgagcgca atggcgttaa aattgaccca aaggttttac
     8041 ataaccactc agaggaactg acgctgcgct tagccgaatt ggagaaaaag gctcacgaga
     8101 tcgctggcga agagttcaat ctgtcatcta cgaaacaact gcagactatc ctgttcgaga
     8161 agcaaggtat caagccatta aaaaagaccc ctggcggtgc tccgtctacc tctgaggaag
     8221 ttttggagga gttagctttg gattaccctc tgccgaaggt tatcttggaa taccgcggtt
     8281 tggctaaatt gaagtctact tatacggata aacttccttt gatgattaat ccaaagacgg
     8341 gtcgcgttca cacgtcgtac catcaagctg ttaccgctac cggtcgcctg tcttctacgg
     8401 atccgaattt acagaatatt cctgtgcgca atgaggaggg ccgccgcatt cgtcaagctt
     8461 ttatcgctcc ggaagactac gttatcgttt ctgctgatta ttctcaaaat gaattacgta
     8521 tcatggctca cctgtctcgc gataagggtc tgttgacggc ctttgctgag ggtaaggaca
     8581 ttcatcgtgc taccgctgct gaggtttacg gcctgccgtt ggaaacggtt acgtctgaac
     8641 agcgtcgctc tgctaagcgt attaatttcg gcttaatcta cggtatgtct gcgtttggct
     8701 tagctcgtca gctgaatatc ccgcgcaagg aagctcaaaa atatatggat ctgtattttg
     8761 agcgttacca cggtgttttg gaatacatgg agcgtacgcg cgcgcaagct aaggaacaag
     8821 gttatgtgga aaccttggat ggtcgtcgct tgtacttgcc tgacattaag tcttctaacg
     8881 gcgcccgccg cgctgctgcc gagcgcgctg ctatcaatgc tccgatgcaa ggtactgctg
     8941 ctgatattat taagcgtgct atgatcgctg tggacgcttg gctgcaagct gaacagcctc
     9001 gcgttcgcat gattatgcaa gttcatgacg agttggtttt cgaggtgcat aaggacgacg
     9061 tggacgctgt tgctaaacaa atccaccagt tgatggagaa ttgcacgcgc ttagacgttc
     9121 cgctgctggt tgaagttggt tctggtgaaa actgggacca ggctcactaa tggctgaaaa
     9181 ctgggaccag gctcactaac tcagatccta ctaggtttaa taaacatctt tattttcatt
     9241 acatctgtgt gttggttttt tgtgtggtac ccaattcgcc ctatagtgag tcgtattaca
     9301 attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     9361 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     9421 atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
     9481 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     9541 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     9601 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     9661 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     9721 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     9781 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     9841 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     9901 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     9961 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
    10021 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
    10081 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
    10141 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
    10201 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
    10261 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
    10321 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
    10381 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
    10441 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
    10501 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
    10561 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
    10621 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
    10681 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
    10741 taaatctgga gccggtgagc gtggttctcg cggtatcatt gcagcactgg ggccagatgg
    10801 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
    10861 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
    10921 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
    10981 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
    11041 ctgagcgtca gaccccgtag aaaagatcaa aggatcttc
//
