pET-PE2-His (IK1822)

LOCUS       pET-PE2-His (IK1822)               11619 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  T7 promoter bacterial expression plasmid for PE2 
            (nSpCas9(H840A)-MMLV-RT) with C-terminal 6xHis-tag and bipartite 
            NLS.
ACCESSION   .
VERSION     .
KEYWORDS    pET-PE2-His (IK1822)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11619)
  TITLE     CRISPR prime editing with ribonucleoprotein complexes in zebrafish 
            and primary human cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 11619)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..11619
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      57..512
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(144..163)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     354..375
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(604..1419)
                     /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     1327..1346
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1411..1430
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1541..2129
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2030..2049
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2283..2300
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2315..2454
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2380..2399)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2540..2562
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2556..2747)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3556..4638)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4600..4619
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4639..4716)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4922..4941
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5025..5044
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5025..5043
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5044..5068
                     /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)."
     RBS             5083..5105
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5148..5168
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5169..9269
                     /codon_start=1
                     /product="nickase mutant of the Cas9 endonuclease from the 
                     Streptococcus pyogenes Type II CRISPR/Cas system"
                     /label=Cas9(H840A)
                     /note="generates RNA-guided single strand nicks in DNA due 
                     to the H840A mutation in the HNH catalytic domain"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             11442..11462
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             11472..11489
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             11508..11525
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(11578..11596)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      11592..20
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc
       61 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
      121 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
      181 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
      241 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
      301 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
      361 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
      421 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
      481 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
      541 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgaattaa
      601 ttcttagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
      661 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
      721 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
      781 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
      841 gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg
      901 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
      961 ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat
     1021 cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg
     1081 atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc
     1141 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
     1201 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
     1261 aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc
     1321 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
     1381 cggcctagag caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt
     1441 tatgtaagca gacagtttta ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc
     1501 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     1561 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     1621 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     1681 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     1741 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     1801 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     1861 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     1921 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     1981 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2041 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     2101 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     2161 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     2221 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     2281 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     2341 atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc
     2401 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     2461 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     2521 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     2581 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
     2641 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
     2701 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
     2761 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
     2821 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
     2881 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     2941 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     3001 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
     3061 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
     3121 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
     3181 ctcaacgaca ggagcacgat catgcgcacc cgtggggccg ccatgccggc gataatggcc
     3241 tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc
     3301 aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc
     3361 tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag
     3421 acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg
     3481 ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca
     3541 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     3601 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt
     3661 tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag
     3721 ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt
     3781 taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc
     3841 cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc
     3901 gttggcaacc agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg
     3961 aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg
     4021 agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc
     4081 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
     4141 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
     4201 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
     4261 atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca
     4321 ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc
     4381 gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc
     4441 aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta
     4501 attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc
     4561 ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta
     4621 taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc
     4681 cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat
     4741 gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg
     4801 caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
     4861 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
     4921 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
     4981 cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg aaattaatac gactcactat
     5041 aggggaattg tgagcggata acaattcccc tctagaaata attttgttta actttaagaa
     5101 ggagatatac catgaaacgg acagccgacg gaagcgagtt cgagtcacca aagaagaagc
     5161 ggaaagtcga taagaaatac tcaataggct tagatatcgg cacaaatagc gtcggatggg
     5221 cggtgatcac tgatgaatat aaggttccgt ctaaaaagtt caaggttctg ggaaatacag
     5281 accgccacag tatcaaaaaa aatcttatag gggctctttt atttgacagt ggagagacag
     5341 cggaagcgac tcgtctcaaa cggacagctc gtagaaggta tacacgtcgg aagaatcgta
     5401 tttgttatct acaggagatt ttttcaaatg agatggcgaa agtagatgat agtttctttc
     5461 atcgacttga agagtctttt ttggtggaag aagacaagaa gcatgaacgt catcctattt
     5521 ttggaaatat agtagatgaa gttgcttatc atgagaaata tccaactatc tatcatctgc
     5581 gaaaaaaatt ggtagattct actgataaag cggatttgcg cttaatctat ttggccttag
     5641 cgcatatgat taagtttcgt ggtcattttt tgattgaggg agatttaaat cctgataata
     5701 gtgatgtgga caaactattt atccagttgg tacaaaccta caatcaatta tttgaagaaa
     5761 accctattaa cgcaagtgga gtagatgcta aagcgattct ttctgcacga ttgagtaaat
     5821 caagacgatt agaaaatctc attgctcagc tccccggtga gaagaaaaat ggcttatttg
     5881 ggaatctcat tgctttgtca ttgggtttga cccctaattt taaatcaaat tttgatttgg
     5941 cagaagatgc taaattacag ctttcaaaag atacttacga tgatgattta gataatttat
     6001 tggcgcaaat tggagatcaa tatgctgatt tgtttttggc agctaagaat ttatcagatg
     6061 ctattttact ttcagatatc ctaagagtaa atactgaaat aactaaggct cccctatcag
     6121 cttcaatgat taaacgctac gatgaacatc atcaagactt gactctttta aaagctttag
     6181 ttcgacaaca acttccagaa aagtataaag aaatcttttt tgatcaatca aaaaacggat
     6241 atgcaggtta tattgatggg ggagctagcc aagaagaatt ttataaattt atcaaaccaa
     6301 ttttagaaaa aatggatggt actgaggaat tattggtgaa actaaatcgt gaagatttgc
     6361 tgcgcaagca acggaccttt gacaacggct ctattcccca tcaaattcac ttgggtgagc
     6421 tgcatgctat tttgagaaga caagaagact tttatccatt tttaaaagac aatcgtgaga
     6481 agattgaaaa aatcttgact tttcgaattc cttattatgt tggtccattg gcgcgtggca
     6541 atagtcgttt tgcatggatg actcggaagt ctgaagaaac aattacccca tggaattttg
     6601 aagaagttgt cgataaaggt gcttcagctc aatcatttat tgaacgcatg acaaactttg
     6661 ataaaaatct tccaaatgaa aaagtactac caaaacatag tttgctttat gagtatttta
     6721 cggtttataa cgaattgaca aaggtcaaat atgttactga aggaatgcga aaaccagcat
     6781 ttctttcagg tgaacagaag aaagccattg ttgatttact cttcaaaaca aatcgaaaag
     6841 taaccgttaa gcaattaaaa gaagattatt tcaaaaaaat agaatgtttt gatagtgttg
     6901 aaatttcagg agttgaagat agatttaatg cttcattagg tacctaccat gatttgctaa
     6961 aaattattaa agataaagat tttttggata atgaagaaaa tgaagatatc ttagaggata
     7021 ttgttttaac attgacctta tttgaagata gggagatgat tgaggaaaga cttaaaacat
     7081 atgctcacct ctttgatgat aaggtgatga aacagcttaa acgtcgccgt tatactggtt
     7141 ggggacgttt gtctcgaaaa ttgattaatg gtattaggga taagcaatct ggcaaaacaa
     7201 tattagattt tttgaaatca gatggttttg ccaatcgcaa ttttatgcag ctgatccatg
     7261 atgatagttt gacatttaaa gaagacattc aaaaagcaca agtgtctgga caaggcgata
     7321 gtttacatga acatattgca aatttagctg gtagccctgc tattaaaaaa ggtattttac
     7381 agactgtaaa agttgttgat gaattggtca aagtaatggg gcggcataag ccagaaaata
     7441 tcgttattga aatggcacgt gaaaatcaga caactcaaaa gggccagaaa aattcgcgag
     7501 agcgtatgaa acgaatcgaa gaaggtatca aagaattagg aagtcagatt cttaaagagc
     7561 atcctgttga aaatactcaa ttgcaaaatg aaaagctcta tctctattat ctccaaaatg
     7621 gaagagacat gtatgtggac caagaattag atattaatcg tttaagtgat tatgatgtcg
     7681 atgcaattgt tccacaaagt ttccttaaag acgattcaat agacaataag gtcttaacgc
     7741 gttctgataa aaatcgtggt aaatcggata acgttccaag tgaagaagta gtcaaaaaga
     7801 tgaaaaacta ttggagacaa cttctaaacg ccaagttaat cactcaacgt aagtttgata
     7861 atttaacgaa agctgaacgt ggaggtttga gtgaacttga taaagctggt tttatcaaac
     7921 gccaattggt tgaaactcgc caaatcacta agcatgtggc acaaattttg gatagtcgca
     7981 tgaatactaa atacgatgaa aatgataaac ttattcgaga ggttaaagtg attaccttaa
     8041 aatctaaatt agtttctgac ttccgaaaag atttccaatt ctataaagta cgtgagatta
     8101 acaattacca tcatgcccat gatgcgtatc taaatgccgt cgttggaact gctttgatta
     8161 agaaatatcc aaaacttgaa tcggagtttg tctatggtga ttataaagtt tatgatgttc
     8221 gtaaaatgat tgctaagtct gagcaagaaa taggcaaagc aaccgcaaaa tatttctttt
     8281 actctaatat catgaacttc ttcaaaacag aaattacact tgcaaatgga gagattcgca
     8341 aacgccctct aatcgaaact aatggggaaa ctggagaaat tgtctgggat aaagggcgag
     8401 attttgccac agtgcgcaaa gtattgtcca tgccccaagt caatattgtc aagaaaacag
     8461 aagtacagac aggcggattc tccaaggagt caattttacc aaaaagaaat tcggacaagc
     8521 ttattgctcg taaaaaagac tgggatccaa aaaaatatgg tggttttgat agtccaacgg
     8581 tagcttattc agtcctagtg gttgctaagg tggaaaaagg gaaatcgaag aagttaaaat
     8641 ccgttaaaga gttactaggg atcacaatta tggaaagaag ttcctttgaa aaaaatccga
     8701 ttgacttttt agaagctaaa ggatataagg aagttaaaaa agacttaatc attaaactac
     8761 ctaaatatag tctttttgag ttagaaaacg gtcgtaaacg gatgctggct agtgccggag
     8821 aattacaaaa aggaaatgag ctggctctgc caagcaaata tgtgaatttt ttatatttag
     8881 ctagtcatta tgaaaagttg aagggtagtc cagaagataa cgaacaaaaa caattgtttg
     8941 tggagcagca taagcattat ttagatgaga ttattgagca aatcagtgaa ttttctaagc
     9001 gtgttatttt agcagatgcc aatttagata aagttcttag tgcatataac aaacatagag
     9061 acaaaccaat acgtgaacaa gcagaaaata ttattcattt atttacgttg acgaatcttg
     9121 gagctccagc cgcattcaag tattttgaca caacgataga tcgcaaacga tacacttcta
     9181 ccaaggaggt gctagacgcg acactgattc accaatccat cacgggatta tatgaaactc
     9241 ggatagattt gtcacagctt gggggtgact ctggaggatc tagcggagga tcctctggca
     9301 gcgagacacc aggaacaagc gagtcagcaa caccagagag cagtggcggc agcagcggcg
     9361 gcagcagcac cctaaatata gaagatgagt atcggctaca tgagacctca aaagagccag
     9421 atgtttctct agggtccaca tggctgtctg attttcctca ggcctgggcg gaaaccgggg
     9481 gcatgggact ggcagttcgc caagctcctc tgatcatacc tctgaaagca acctctaccc
     9541 ccgtgtccat aaaacaatac cccatgtcac aagaagccag actggggatc aagccccaca
     9601 tacagagact gttggaccag ggaatactgg taccctgcca gtccccctgg aacacgcccc
     9661 tgctacccgt taagaaacca gggactaatg attataggcc tgtccaggat ctgagagaag
     9721 tcaacaagcg ggtggaagac atccacccca ccgtgcccaa cccttacaac ctcttgagcg
     9781 ggctcccacc gtcccaccag tggtacactg tgcttgattt aaaggatgcc tttttctgcc
     9841 tgagactcca ccccaccagt cagcctctct tcgcctttga gtggagagat ccagagatgg
     9901 gaatctcagg acaattgacc tggaccagac tcccacaggg tttcaaaaac agtcccaccc
     9961 tgtttaatga ggcactgcac agagacctag cagacttccg gatccagcac ccagacttga
    10021 tcctgctaca gtacgtggat gacttactgc tggccgccac ttctgagcta gactgccaac
    10081 aaggtactcg ggccctgtta caaaccctag ggaacctcgg gtatcgggcc tcggccaaga
    10141 aagcccaaat ttgccagaaa caggtcaagt atctggggta tcttctaaaa gagggtcaga
    10201 gatggctgac tgaggccaga aaagagactg tgatggggca gcctactccg aagacccctc
    10261 gacaactaag ggagttccta gggaaggcag gcttctgtcg cctcttcatc cctgggtttg
    10321 cagaaatggc agcccccctg taccctctca ccaaaccggg gactctgttt aattggggcc
    10381 cagaccaaca aaaggcctat caagaaatca agcaagctct tctaactgcc ccagccctgg
    10441 ggttgccaga tttgactaag ccctttgaac tctttgtcga cgagaagcag ggctacgcca
    10501 aaggtgtcct aacgcaaaaa ctgggacctt ggcgtcggcc ggtggcctac ctgtccaaaa
    10561 agctagaccc agtagcagct gggtggcccc cttgcctacg gatggtagca gccattgccg
    10621 tactgacaaa ggatgcaggc aagctaacca tgggacagcc actagtcatt ctggcccccc
    10681 atgcagtaga ggcactagtc aaacaacccc ccgaccgctg gctttccaac gcccggatga
    10741 ctcactatca ggccttgctt ttggacacgg accgggtcca gttcggaccg gtggtagccc
    10801 tgaacccggc tacgctgctc ccactgcctg aggaagggct gcaacacaac tgccttgata
    10861 tcctggccga agcccacgga acccgacccg acctaacgga ccagccgctc ccagacgccg
    10921 accacacctg gtacacggat ggaagcagtc tcttacaaga gggacagcgt aaggcgggag
    10981 ctgcggtgac caccgagacc gaggtaatct gggctaaagc cctgccagcc gggacatccg
    11041 ctcagcgggc tgaactgata gcactcaccc aggccctaaa gatggcagaa ggtaagaagc
    11101 taaatgttta tactgatagc cgttatgctt ttgctactgc ccatatccat ggagaaatat
    11161 acagaaggcg tgggtggctc acatcagaag gcaaagagat caaaaataaa gacgagatct
    11221 tggccctact aaaagccctc tttctgccca aaagacttag cataatccat tgtccaggac
    11281 atcaaaaggg acacagcgcc gaggctagag gcaaccggat ggctgaccaa gcggcccgaa
    11341 aggcagccat cacagagact ccagacacct ctaccctcct catagaaaat tcatcaccct
    11401 ctggcggctc aaaaagaacc gccgacggca gcgaattcga gcccaagaag aagaggaaag
    11461 tcggcagcag ccatcatcat catcatcact aagcggccgc actcgagcac caccaccacc
    11521 accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
    11581 ctgagcaata actagcataa ccccttgggg cctctaaac
//