pEvolvR-enCas9-PolI5M

LOCUS       pEvolvR-enCas9-PolI5M              12546 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses enCas9 fused to PolI5M in E. coli. contains a 
            BsmbI-flanked GFP cassette for sgRNA cloning..
ACCESSION   .
VERSION     .
KEYWORDS    pEvolvR-enCas9-PolI5M
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12546)
  AUTHORS   Halperin SO, Tou CJ, Wong EB, Modavi C, Schaffer DV, Dueber JE
  TITLE     CRISPR-guided DNA polymerases enable diversification of all 
            nucleotides in a tunable window.
  JOURNAL   Nature. 2018 Aug 1. pii: 10.1038/s41586-018-0384-8. doi: 
            10.1038/s41586-018-0384-8.
  PUBMED    30069054
REFERENCE   2  (bases 1 to 12546)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..12546
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     RBS             4884..4892
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             complement(7094..7909)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     7817..7836
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7901..7920
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      8138..8726
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8627..8646
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8880..8897
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8912..9051
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8977..8996)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9137..9159
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(9153..9344)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     RBS             9474..9485
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             9492..10151
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKF*PAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(9558..9577)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     RBS             10451..10462
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             10469..11182
                     /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      11194..11265
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      11281..11308
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_RNA        11322..11397
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             11418..11447
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             11463..11480
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(11536..11553)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      11706..11752
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(11811..12434)
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     12342..12361
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        12453..12508
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    12489..12507
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             12526..12537
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 gacaagaagt attctatcgg actggctatc gggactaata gcgtcgggtg ggccgtgatc
       61 actgacgagt acaaggtgcc ctctaagaag ttcaaggtgc tcgggaacac cgaccggcat
      121 tccatcaaga aaaatctgat cggagctctc ctctttgatt caggggaaac cgctgaagca
      181 acccgcctca agcggactgc tagacggcgg tacaccagga ggaagaaccg gatttgttac
      241 cttcaagaga tattctccaa cgaaatggca aaggtcgacg acagcttctt ccataggctg
      301 gaagaatcat tcctcgtgga agaggataag aagcatgaac ggcatcccat cttcggtaat
      361 atcgtcgacg aggtggccta tcacgagaaa tacccaacca tctaccatct tcgcaaaaag
      421 ctggtggact caaccgacaa ggcagacctc cggcttatct acctggccct ggcccacatg
      481 atcaagttca gaggccactt cctgatcgag ggcgacctca atcctgacaa tagcgatgtg
      541 gataaactgt tcatccagct ggtgcagact tacaaccagc tctttgaaga gaaccccatc
      601 aatgcaagcg gagtcgatgc caaggccatt ctgtcagccc ggctgtcaaa gagccgcaga
      661 cttgagaatc ttatcgctca gctgccgggt gaaaagaaaa atggactgtt cgggaacctg
      721 attgctcttt cacttgggct gactcccaat ttcaagtcta atttcgacct ggcagaggat
      781 gccaagctgc aactgtccaa ggacacctat gatgacgatc tcgacaacct cctggcccag
      841 atcggtgacc aatacgccga ccttttcctt gctgctaaga atctttctga cgccatcctg
      901 ctgtctgaca ttctccgcgt gaacactgaa atcaccaagg cccctctttc agcttcaatg
      961 attaagcggt atgatgagca ccaccaggac ctgaccctgc ttaaggcact cgtccggcag
     1021 cagcttccgg agaagtacaa ggaaatcttc tttgaccagt caaagaatgg atacgccggc
     1081 tacatcgacg gaggtgcctc ccaagaggaa ttttataagt ttatcaaacc tatccttgag
     1141 aagatggacg gcaccgaaga gctcctcgtg aaactgaatc gggaggatct gctgcggaag
     1201 cagcgcactt tcgacaatgg gagcattccc caccagatcc atcttgggga gcttcacgcc
     1261 atccttcggc gccaagagga cttctacccc tttcttaagg acaacaggga gaagattgag
     1321 aaaattctca ctttccgcat cccctactac gtgggacccc tcgccagagg aaatagccgg
     1381 tttgcttgga tgaccagaaa gtcagaagaa actatcactc cctggaactt cgaagaggtg
     1441 gtggacaagg gagccagcgc tcagtcattc atcgaacgga tgactaactt cgataagaac
     1501 ctccccaatg agaaggtcct gccgaaacat tccctgctct acgagtactt taccgtgtac
     1561 aacgagctga ccaaggtgaa atatgtcacc gaagggatga ggaagcccgc attcctgtca
     1621 ggcgaacaaa agaaggcaat tgtggacctt ctgttcaaga ccaatagaaa ggtgaccgtg
     1681 aagcagctga aggaggacta tttcaagaaa attgaatgct tcgactctgt ggagattagc
     1741 ggggtcgaag atcggttcaa cgcaagcctg ggtacttacc atgatctgct taagatcatc
     1801 aaggacaagg attttctgga caatgaggag aacgaggaca tccttgagga cattgtcctg
     1861 actctcactc tgttcgagga ccgggaaatg atcgaggaga ggcttaagac ctacgcccat
     1921 ctgttcgacg ataaagtgat gaagcaactt aaacggagaa gatataccgg atggggacgc
     1981 cttagccgca aactcatcaa cggaatccgg gacaaacaga gcggaaagac cattcttgat
     2041 ttccttaaga gcgacggatt cgctaatcgc aacttcatgc aacttatcca tgatgattcc
     2101 ctgaccttta aggaggacat ccagaaggcc caagtgtctg gacaaggtga ctcactgcac
     2161 gagcatatcg caaatctggc tggttcaccc gctattaaga agggtattct ccagaccgtg
     2221 aaagtcgtgg acgagctggt caaggtgatg ggtcgccata aaccagagaa cattgtcatc
     2281 gagatggcca gggaaaacca gactacccag aagggacaga agaacagcag ggagcggatg
     2341 aaaagaattg aggaagggat taaggagctc gggtcacaga tccttaaaga gcacccggtg
     2401 gaaaacaccc agcttcagaa tgagaagctc tatctgtact accttcaaaa tggacgcgat
     2461 atgtatgtgg accaagagct tgatatcaac aggctctcag actacgacgt ggaccatatc
     2521 gtccctcaga gcttcctcgc agacgactca attgacaata aggtgctgac tcgctcagac
     2581 aagaaccggg gaaagtcaga taacgtgccc tcagaggaag tcgtgaaaaa gatgaagaac
     2641 tattggcgcc agcttctgaa cgcaaagctg atcactcagc ggaagttcga caatctcact
     2701 aaggctgaga ggggcggact gagcgaactg gacaaagcag gattcattaa acggcaactt
     2761 gtggagactc ggcagattac taaacatgtc gcccaaatcc ttgactcacg catgaatacc
     2821 aagtacgacg aaaacgacaa acttatccgc gaggtgaagg tgattaccct gaagtccaag
     2881 ctggtcagcg atttcagaaa ggactttcaa ttctacaaag tgcgggagat caataactat
     2941 catcatgctc atgacgcata tctgaatgcc gtggtgggaa ccgccctgat caagaagtac
     3001 ccagcactgg aaagcgagtt cgtgtacgga gactacaagg tctacgacgt gcgcaagatg
     3061 attgccaaat ctgagcagga gatcggaaag gccaccgcaa agtacttctt ctacagcaac
     3121 atcatgaatt tcttcaagac cgaaatcacc cttgcaaacg gtgagatccg gaaggcgccg
     3181 ctcatcgaga ctaatgggga gactggcgaa atcgtgtggg acaagggcag agatttcgct
     3241 accgtgcgca aagtgctttc tatgcctcaa gtgaacatcg tgaagaaaac cgaggtgcaa
     3301 accggaggct tttctaagga atcaatcctc cccaagcgca actccgacaa gctcattgca
     3361 aggaagaagg attgggaccc taagaagtac ggcggattcg attcaccaac tgtggcttat
     3421 tctgtcctgg tcgtggctaa ggtggaaaaa ggaaagtcta agaagctcaa gagcgtgaag
     3481 gaactgctgg gtatcaccat tatggagcgc agctccttcg agaagaaccc aattgacttt
     3541 ctcgaagcca aaggttacaa ggaagtcaag aaggacctta tcatcaagct cccaaagtat
     3601 agcctgttcg aactggagaa tgggcggaag cggatgctcg cctccgctgg cgaacttcag
     3661 aagggtaatg agctggctct cccctccaag tacgtgaatt tcctctacct tgcaagccat
     3721 tacgagaagc tgaaggggag ccccgaggac aacgagcaaa agcaactgtt tgtggagcag
     3781 cataagcatt atctggacga gatcattgag cagatttccg agttttctaa acgcgtcatt
     3841 ctcgctgatg ccaacctcga taaagtcctt agcgcataca ataagcacag agacaaacca
     3901 attcgggagc aggctgagaa tatcatccac ctgttcaccc tcaccaatct tggtgcccct
     3961 gccgcattca agtacttcga caccaccatc gaccggaaac gctatacctc caccaaagaa
     4021 gtgctggacg ccaccctcat ccaccagagc atcaccggac tttacgaaac tcggattgac
     4081 ctctcacagc tcggagggga tggttctagt gaaaccccgg gaacaagtga gtcggccacc
     4141 cctgaaggtg gatcaggggg tagcggatcc gttcagatcc cgcagaaccc gctgattctg
     4201 gttgacggat ctagttacct gtaccgtgct taccatgctt tcccgccttt gaccaattct
     4261 gctggtgaac ctacgggagc tatgtacgga gttctgaata tgttgcgttc tttaattatg
     4321 cagtacaagc ctacccacgc tgctgttgtt ttcgatgcta aaggtaagac gttccgcgac
     4381 gagttattcg agcactataa gtctcaccgt cctccgatgc ctgatgactt acgcgctcag
     4441 attgagccgc tgcatgctat ggtgaaggct atgggtttac ctcttttggc tgtcagcggt
     4501 gttgaggctg atgatgtcat tggcacctta gctcgtgagg ctgagaaggc tggtcgccct
     4561 gttttgattt ctaccggtga caaggacatg gctcaattgg ttaccccgaa catcaccctg
     4621 atcaacacca tgaccaacac gattctgggt cctgaggaag ttgttaacaa atatggtgtt
     4681 cctccggagt tgattattga ctttcttgct cttatgggcg attcttcaga caatatcccg
     4741 ggtgttccag gtgttggaga gaagactgct caagctctgc ttcagggtct gggtggtttg
     4801 gacacccttt acgctgaacc ggagaagatc gccggtctgt cttttcgcgg tgctaagacc
     4861 atggctgcta aactggaaca gaataaggag gtcgcatacc tgtcttatca attggctacc
     4921 atcaagacgg atgtggagtt agaacttacg tgcgagcagc ttgaggttca acagcctgct
     4981 gctgaggaac tgctgggtct ttttaagaaa tacgaattta agcgttggac cgccgacgtt
     5041 gaggctggta agtggctgca agctaagggt gctaagccgg ctgctaaacc gcaagaaacg
     5101 agtgtcgctg atgaggctcc ggaggttacc gctaccgtta tctcttacga taattatgtt
     5161 acgattctgg acgaggaaac cttaaaggct tggatcgcta aattagagaa ggctcctgtt
     5221 ttcgctttcg acacggaaac ggattctctg gacaatatta gtgcgaatct tgttggtctg
     5281 agtttcgcaa ttgaaccggg tgttgctgct tacatccctg tggcacacga ctacctggac
     5341 gctccggacc agatttcacg tgaacgcgct ctggaactgc tgaagccttt attagaggac
     5401 gagaaagctt tgaaagttgg tcagaatttg aagtatgctc gtggaatctt agctaattat
     5461 ggtatcgagt tgcgcggtat cgctttcgac acgatgttgg aatcttatat cctgaactct
     5521 gtcgctggtc gccatgacat ggactctctg gctgagcgct ggctgaaaca taagacgatt
     5581 accttcgagg aaatcgcagg aaagggtaag aaccagctca cgttcaatca aatcgctctg
     5641 gaggaagctg gtcgctatgc tgctgaggac gctgacgtta ctctgcaact gcacttgaag
     5701 atgtggcctg acttgcagaa gcataagggt ccactgaatg tttttgaaaa cattgagatg
     5761 cctttggttc cagttctgtc tcgtatcgag cgcaatggcg ttaaaattga cccaaaggtt
     5821 ttacataacc actcagagga actgacgctg cgcttagccg aattggagaa aaaggctcac
     5881 gagatcgctg gcgaagagtt caatctgtca tctacgaaac aactgcagac tatcctgttc
     5941 gagaagcaag gtatcaagcc attaaaaaag acccctggcg gtgctccgtc tacctctgag
     6001 gaagttttgg aggagttagc tttggattac cctctgccga aggttatctt ggaataccgc
     6061 ggtttggcta aattgaagtc tacttatacg gataaacttc ctttgatgat taatccaaag
     6121 acgggtcgcg ttcacacgtc gtaccatcaa gctgttaccg ctaccggtcg cctgtcttct
     6181 acggatccga atttacagaa tattcctgtg cgcaatgagg agggccgccg cattcgtcaa
     6241 gcttttatcg ctccggaaga ctacgttatc gtttctgctg attattctca aaatgaatta
     6301 cgtatcatgg ctcacctgtc tcgcgataag ggtctgttga cggcctttgc tgagggtaag
     6361 gacattcatc gtgctaccgc tgctgaggtt tacggcctgc cgttggaaac ggttacgtct
     6421 gaacagcgtc gctctgctaa gcgtattaat ttcggcttaa tctacggtat gtctgcgttt
     6481 ggcttagctc gtcagctgaa tatcccgcgc aaggaagctc aaaaatatat ggatctgtat
     6541 tttgagcgtt accacggtgt tttggaatac atggagcgta cgcgcgcgca agctaaggaa
     6601 caaggttatg tggaaacctt ggatggtcgt cgcttgtact tgcctgacat taagtcttct
     6661 aacggcgccc gccgcgctgc tgccgagcgc gctgctatca atgctccgat gcaaggtact
     6721 gctgctgata ttattaagcg tgctatgatc gctgtggacg cttggctgca agctgaacag
     6781 cctcgcgttc gcatgattat gcaagttcat gacgagttgg ttttcgaggt gcataaggac
     6841 gacgtggacg ctgttgctaa acaaatccac cagttgatgg agaattgcac gcgcttagac
     6901 gttccgctgc tggttgaagt tggttctggt gaaaactggg accaggctca ctaataatgg
     6961 cctcggtacc aaagacgaac aataagacgc tgaaaagcgt cttttttcgt tttggtccgc
     7021 tgagcagtta cagagatgtt acgaaccact agtgcactgc agtacagtgt tacaaccaat
     7081 taaccaattc tgattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat
     7141 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac
     7201 cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa
     7261 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac
     7321 catgagtgac gactgaatcc ggtgagaatg gcaaaagctt atgcatttct ttccagactt
     7381 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat
     7441 tcattcgtga ttgcgcctga gcgaggcgaa atacgcgatc gctgttaaaa ggacaattac
     7501 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac
     7561 ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga
     7621 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt
     7681 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc
     7741 catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac
     7801 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg
     7861 aatttaatcg cggcctggag caagacgttt cccgttgaat atggctcata acaccccttg
     7921 tattactgtt tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg
     7981 caatgtaaca tcagagattt tgagacacaa cgtggctttg ttgaataaat cgaacttttg
     8041 ctgagttgaa ggatcaggag ttcatgacca aaatccctta acgtgagttt tcgttccact
     8101 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     8161 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     8221 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     8281 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     8341 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     8401 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     8461 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     8521 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     8581 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     8641 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     8701 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     8761 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     8821 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     8881 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     8941 tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata
     9001 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     9061 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     9121 caagctgtga ccttctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     9181 cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct
     9241 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg
     9301 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa
     9361 gggggatttc tgttcatccc tgaattcgca tctagactga actggccgat aattgcagac
     9421 gaacggattt actttacagc tagctcagtc ctaggtatta tgctagcgct agcaaagagg
     9481 agaaaagatc tatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc
     9541 atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg
     9601 ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttaac
     9661 cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattt cgtatggcaa
     9721 tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg
     9781 agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc
     9841 tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag
     9901 ggtttattga gaatatgttt ttcgtttcag ccaatccctg ggtgagtttc accagttttg
     9961 atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt
    10021 atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg
    10081 atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg
    10141 gcggggcgta aggatcctaa ctcgagctcg gtaccaaatt ccagaaaaga ggcctcccga
    10201 aaggggggcc ttttttcgtt ttggtccgct gccaaaccag atgtcaacac agctacaacg
    10261 tttatagcta gctcagccct tggtacaatg ctagctgaga cggaaagtga aacgtgattt
    10321 catgcgtcat tttgaacatt ttgtaaatct tatttaataa tgtgtgcggc aattcacatt
    10381 taatttatga atgttttctt aacatcgcgg caactcaaga aacggcaggt tcggatctta
    10441 gctactagag aaagaggaga aatactagat gcgtaaaggc gaagagctgt tcactggtgt
    10501 cgtccctatt ctggtggaac tggatggtga tgtcaacggt cataagtttt ccgtgcgtgg
    10561 cgagggtgaa ggtgacgcaa ctaatggtaa actgacgctg aagttcatct gtactactgg
    10621 taaactgccg gttccttggc cgactctggt aacgacgctg acttatggtg ttcagtgctt
    10681 tgctcgttat ccggaccata tgaagcagca tgacttcttc aagtccgcca tgccggaagg
    10741 ctatgtgcag gaacgcacga tttcctttaa ggatgacggc acgtacaaaa cgcgtgcgga
    10801 agtgaaattt gaaggcgata ccctggtaaa ccgcattgag ctgaaaggca ttgactttaa
    10861 agaggacggc aatatcctgg gccataagct ggaatacaat tttaacagcc acaatgttta
    10921 catcaccgcc gataaacaaa aaaatggcat taaagcgaat tttaaaattc gccacaacgt
    10981 ggaggatggc agcgtgcagc tggctgatca ctaccagcaa aacactccaa tcggtgatgg
    11041 tcctgttctg ctgccagaca atcactatct gagcacgcaa agcgttctgt ctaaagatcc
    11101 gaacgagaaa cgcgatcata tggttctgct ggagttcgta accgcagcgg gcatcacgca
    11161 tggtatggat gaactgtaca aatgaccagg catcaaataa aacgaaaggc tcagtcgaaa
    11221 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta gagtcacact
    11281 ggctcacctt cgggtgggcc tttctgcgtt tatacgtctc agttttagag ctagaaatag
    11341 caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt
    11401 ttttgaagct tgggcccgaa caaaaactca tctcagaaga ggatctgaat agcgccgtcg
    11461 accatcatca tcatcatcat tgagtttaaa cggactccag cttggctgtt ttggcggatg
    11521 agagaagatt ttcagcctga tacagattaa atcagaacgc agaagcggtc tgataaaaca
    11581 gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt
    11641 gaaacgccgt agcgccgatg gtagtgtggg gactccccat gcgagagtag ggaactgcca
    11701 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
    11761 tgtcggtgaa cgacagggct gccaaaccag atgtcaacac agctacaacg ttaagaccca
    11821 ctttcacatt taagttgttt ttctaatccg catatgatca attcaaggcc gaataagaag
    11881 gctggctctg caccttggtg atcaaataat tcgatagctt gtcgtaataa tggcggcata
    11941 ctatcagtag taggtgtttc cctttcttct ttagcgactt gatgctcttg atcttccaat
    12001 acgcaaccta aagtaaaatg ccccacagcg ctgagtgcat ataatgcatt ctctagtgaa
    12061 aaaccttgtt ggcataaaaa ggctaattga ttttcgagag tttcatactg tttttctgta
    12121 ggccgtgtac ctaaatgtac ttttgctcca tcgcgatgac ttagtaaagc acatctaaaa
    12181 cttttagcgt tattacgtaa aaaatcttgc cagctttccc cttctaaagg gcaaaagtga
    12241 gtatggtgcc tatctaacat ctcaatggct aaggcgtcga gcaaagcccg cttatttttt
    12301 acatgccaat acaatgtagg ctgctctaca cctagcttct gggcgagttt acgggttgtt
    12361 aaaccttcga ttccgacctc attaagcagc tctaatgcgc tgttaatcac tttactttta
    12421 tctaatctag acatcattaa ttcctaattt ttgttgacac tctatcgttg atagagttat
    12481 tttaccactc cctatcagtg atagagaaaa gaattcaaaa gatctaaaga ggagaaaaga
    12541 tctatg
//