LOCUS       His-CL7_MBP_1xNL        8758 bp DNA     circular SYN 16-JAN-2023
DEFINITION  obtaining full Cas9 RNP.
ACCESSION   .
VERSION     .
KEYWORDS    pCold CL7-Cas9
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8758)
  AUTHORS   Qiao J, Li W, Lin S, Sun W, Ma L, Liu Y
  TITLE     Co-expression of Cas9 and single-guided RNAs in Escherichia coli 
            streamlines production of Cas9 ribonucleoproteins.
  JOURNAL   Commun Biol. 2019 May 3;2:161. doi: 10.1038/s42003-019-0402-x. 
            eCollection 2019.
  PUBMED    31069270
REFERENCE   2  (bases 1 to 8758)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 18, 2024 from SnapGene 7.0.2
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Commun 
            Biol."; date: "2019-05-3"; volume: "2:161. doi"; pages: " 
            10.1038/s42003-019-0402-x. eCollection 2019"
FEATURES             Location/Qualifiers
     source          1..8758
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        75..103
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    109..125
                     /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     114..135
                     /label=seq1
     5'UTR           147..280
                     /gene="cspA"
                     /label=cspA 5'UTR
                     /note="5'UTR of the E. coli cold shock protein cspA gene 
                     (Mitta et al., 1997)"
     primer_bind     complement(257..311)
                     /label=His-start-5UTR-rev
     CDS             281..292
                     /codon_start=1
                     /label=start
                     /translation="MKSS"
     primer_bind     288..342
                     /label=His-CL7-fwd
     CDS             293..310
                     /codon_start=1
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             317..721
                     /codon_start=1
                     /label=CL7
                     /translation="SKSNEPGKATGEGKPVNNKWLNNAGKDLGSPVPDRIANKLRDKEF
                     ESFDDFRETFWEEVSKDPELSKQFSRNNNDRMKVGKAPKTRTQDVSGKRTSFELNHQKP
                     IEQNGGVYDMDNISVVTPKRNIDIEGGGGGS"
     primer_bind     complement(693..740)
                     /label=CL7-MBP-rev
     primer_bind     709..757
                     /label=CL7-MBP-fwd
     CDS             731..1831
                     /codon_start=1
                     /label=MBP
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     complement(1860..1908)
                     /label=HRV3C-MBPlin-rev
     primer_bind     complement(1886..1944)
                     /label=Geo-st-SV40-HRV-rev
     CDS             1886..1909
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     1886..1909
                     /label=seq2
     CDS             1916..1936
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     1924..1975
                     /label=SV40-Geo-start-fwd
     CDS             1952..5212
                     /codon_start=1
                     /label=GeoCas9-R1
                     /translation="MRYKIGLDIGITSVGWAVMNLDIPRIEDLGVRIFDRAENPQTGES
                     LALPRRLARSARRRLRRRKHRLERIRRLVIREGILTKEELDKLFEEKHEIDVWQLRVEA
                     LDRKLNNDELARVLLHLAKRRGFKSNRKSERSNKENSTMLKHIEGNRAILSSYRTVGEM
                     IVKDPKFALHKRNKGENYINTIARDDLEREIRLIFSKQREFGDMSCTEEFENEYITIWA
                     SQRPVASKDDIEKKVGFCTFEPKEKRAPKATYTFQSFIAWEHINKLRLISPSGARGLTD
                     EERRLLYEQAFQKNKITYHDIRTLLHLPDDTYFKGIVYDRGESRKQNENIRFLELDAYH
                     QIRKAVDKVYGKGKSSSFLPIDFDTFGYALTLFKDDADIHSYLRNEYEQNGKRMPNLAN
                     KVYDNELIEELLNLSFTKFGHLSLKALRSILPYMEQGEVYSSACERAGYTFTGPKKKQK
                     TMLLPNIQPIANPVVMRALTQARKVVNAIIKKYGSPVSIHIELARDLSQTFDERRKTKK
                     EQDENRKKNETAIRQLMEYGLTLNPTGHDIVKFKLWSEQNGRCAYSLQPIEIERLLEPG
                     YVEVDHVIPYSRSLDDSYTNKVLVLTRENREKGNRIPAEYLGVGTERWQQFETFVLTNK
                     QFSKKKRDRLLRLHYDENEETEFKNRNLNDTRYISRFFANFIREHLKFAESDDKQKVYT
                     VNGRVTAHLRSRWEFNKNREESDLHHAVDAVIVACTTPSDIAKVTAFYQRREQNKELAK
                     KTEPHFPQPWPHFADELRARLSKHPKESIKALNLGNYDDQKLESLQPVFVSRMPKRSVT
                     GAAHRETLRRYVGIDERSGKIQTVVKTKLSKIKLDASGHFPMYGKESDPRTYEAIRQRL
                     LEHNNDPKKAFQGPLYKPKKNGEPGPVIRTVKIIDTRNQVIPLNDGKTVAYNSNIVRVD
                     VFEKDGKYYCVPVYTMDIMKGILPNKAIEPNKPYSEWKEMTEDYTFRFSLYPNDLIRIE
                     LPREKTVKTAAGEEINVKDVFVYYKTIDSANGGLELISHDHRFSLRGVGSRTLKRFEKY
                     QVDVLGNIYKVRGEKRVGLASSAHSKPGKTIRPLQSTRD"
     misc_feature    1952..2095
                     /label=RuvC-I
     primer_bind     1952..1975
                     /label=Geo-start-fwd
     primer_bind     2073..2094
                     /label=Rec-start-fwd
     primer_bind     complement(2073..2094)
                     /label=Rec-start-rev
     misc_feature    2093..2095
                     /label=L48L
     misc_feature    2096..3328
                     /label=Rec lobe
     misc_feature    2396..2398
                     /label=E149G
     misc_feature    2495..2497
                     /label=T182I
     misc_feature    2567..2569
                     /label=N206D
     primer_bind     3326..3347
                     /label=Rec-end-fwd
     primer_bind     complement(3326..3347)
                     /label=Rec-end-rev
     misc_feature    3329..3484
                     /label=RuvC-II
     misc_feature    3347..3349
                     /label=P466Q
     primer_bind     3410..3432
                     /label=seq-GeoHNH-fwd
     primer_bind     complement(3410..3432)
                     /label=seq-GeoRec-rev
     misc_feature    3485..3937
                     /label=HNH
     misc_feature    3938..4441
                     /label=RuvC-III
     primer_bind     4353..4376
                     /label=seq-GeoPI-fwd
     primer_bind     complement(4353..4376)
                     /label=seq-GeoHNH-rev
     misc_feature    4400..4402
                     /label=Q817R
     primer_bind     4415..4437
                     /label=PI-start-fwd-2
     primer_bind     complement(4415..4437)
                     /label=PI-start-rev-2
     primer_bind     4415..4436
                     /label=PI-start-fwd
     primer_bind     complement(4415..4436)
                     /label=PI-start-rev
     misc_feature    4442..5212
                     /label=PAM Interacting
     primer_bind     4457..4498
                     /label=Geo-E843K-fwd
     misc_feature    4478..4480
                     /label=E843K
     misc_feature    4601..4603
                     /label=E884G
     misc_feature    4673..4675
                     /label=K908R
     primer_bind     5075..5096
                     /label=seq-G1
     primer_bind     complement(5188..5212)
                     /label=Geo-end-rev
     CDS             5219..5239
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     5242..5298
                     /label=SV40-3UTR-fwd
     primer_bind     complement(5242..5269)
                     /label=SV40-rev
     CDS             5249..5269
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     3'UTR           5280..5424
                     /gene="cspA"
                     /label=cspA 3'UTR
                     /note="3'UTR of the E. coli cold shock protein cspA gene 
                     (Mitta et al., 1997)"
     terminator      5500..5571
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        5677..5781
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5782..6642
                     /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(6000..6019)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6813..7401
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7302..7321
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7573..8655)
                     /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     8617..8636
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(8656..8733)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc
       61 aaggaatggt gtggttgaca attaatcatc ggctcgtata atgtgtaatt gtgagcggat
      121 aacaatttga tgtgctagcg catatccagt gtagtaaggc aagtcccttc aagagttatc
      181 gttgataccc ctcgtagtgc acattccttt aacgcttcaa aatctgtaaa gcacgccata
      241 tcgccgaaag gcacacttaa ttattaagag gtaatacacc atgaaatctt ctcaccatca
      301 ccatcaccat ggttctagca aaagcaatga accgggtaaa gcaaccggtg aaggtaaacc
      361 ggttaataac aaatggctga acaatgccgg taaagatctg ggtagtccgg ttccggatcg
      421 tattgcaaat aaactgcgtg ataaagaatt cgagagcttc gatgattttc gtgaaacctt
      481 ttgggaagaa gttagcaaag atcctgaact gagcaaacag tttagccgca ataacaatga
      541 tcgtatgaaa gttggtaaag caccgaaaac acgtacccag gatgttagcg gtaaacgtac
      601 ctcatttgaa ctgaatcatc agaaaccgat tgaacagaat ggtggcgttt atgatatgga
      661 taacattagc gttgttaccc cgaaacgcaa cattgatatt gaaggcggtg gtggtggttc
      721 tggttcttct atgaaaatcg aagaaggtaa actggtaatc tggattaacg gcgataaagg
      781 ctataacggt ctcgctgaag tcggtaagaa attcgagaaa gataccggaa ttaaagtcac
      841 cgttgagcat ccggataaac tggaagagaa attcccacag gttgcggcaa ctggcgatgg
      901 ccctgacatt atcttctggg cacacgaccg ctttggtggc tacgctcaat ctggcctgtt
      961 ggctgaaatc accccggaca aagcgttcca ggacaagctg tatccgttta cctgggatgc
     1021 cgtacgttac aacggcaagc tgattgctta cccgatcgct gttgaagcgt tatcgctgat
     1081 ttataacaaa gatctgctgc cgaacccgcc aaaaacctgg gaagagatcc cggcgctgga
     1141 taaagaactg aaagcgaaag gtaagagcgc gctgatgttc aacctgcaag aaccgtactt
     1201 cacctggccg ctgattgctg ctgacggggg ttatgcgttc aagtatgaaa acggcaagta
     1261 cgacattaaa gacgtgggcg tggataacgc tggcgcgaaa gcgggtctga ccttcctggt
     1321 tgacctgatt aaaaacaaac acatgaatgc agacaccgat tactccatcg cagaagctgc
     1381 ctttaataaa ggcgaaacag cgatgaccat caacggcccg tgggcatggt ccaacatcga
     1441 caccagcaaa gtgaattatg gtgtaacggt actgccgacc ttcaagggtc aaccatccaa
     1501 accgttcgtt ggcgtgctga gcgcaggtat taacgccgcc agtccgaaca aagagctggc
     1561 aaaagagttc ctcgaaaact atctgctgac tgatgaaggt ctggaagcgg ttaataaaga
     1621 caaaccgctg ggtgccgtag cgctgaagtc ttacgaggaa gagttggcga aagatccacg
     1681 tattgccgcc actatggaaa acgcccagaa aggtgaaatc atgccgaaca tcccgcagat
     1741 gtccgctttc tggtatgccg tgcgtactgc ggtgatcaac gccgccagcg gtcgtcagac
     1801 tgtcgatgaa gccctgaaag acgcgcagac taattcgagc tcgaacaaca acaacaataa
     1861 caataacaac aacctcggga tcgagttgga ggttttgttc cagggtccag gatctccgaa
     1921 aaagaaacgg aaagtaggcg gttcgaatgc aatgcgttat aagattggcc tggacatcgg
     1981 tattacctct gttggttggg cagtcatgaa cctggatatc cctcgtatcg aagatctggg
     2041 cgtgcgcatt tttgaccgtg cggagaaccc gcagaccggt gaatctctgg ctctaccgcg
     2101 tcgtctggca cgtagcgcac gccgccgcct gcgtcgtcgt aaacaccgtc tggagcgtat
     2161 tcgtcgcctg gttattcgtg aaggcatcct gacgaaagaa gaactggata aactgttcga
     2221 agaaaaacac gagatcgacg tatggcagct gcgtgtagaa gccctggacc gtaagctgaa
     2281 caacgacgaa ctggcgcgtg tcctgctgca tctggcaaag cgtcgtggct tcaaatctaa
     2341 ccgtaaatct gaacgctcca ataaagagaa ctccactatg ctgaaacata ttgaggggaa
     2401 ccgtgcaatt ctgtctagct accgtaccgt gggcgaaatg attgttaaag acccgaaatt
     2461 cgcactgcat aagcgtaaca aaggcgaaaa ctacatcaac accattgcac gcgatgacct
     2521 ggaacgtgaa atccgtctga ttttctccaa acagcgcgaa ttcggcgaca tgtcttgcac
     2581 cgaagaattc gaaaacgaat atattaccat ttgggcatct cagcgtccgg tggcgtctaa
     2641 agatgatatc gaaaaaaaag taggcttttg tactttcgaa ccgaaggaaa aacgtgcgcc
     2701 gaaagccacc tataccttcc agtcttttat cgcgtgggaa catatcaaca aactgcgtct
     2761 gatttctccg tctggcgccc gcggcctgac cgacgaagaa cgtcgtctgc tgtatgaaca
     2821 agcattccag aaaaacaaaa ttacctacca cgatattcgt accctgctgc atctgccgga
     2881 cgacacctac ttcaagggca tcgtttacga tcgcggtgaa tctcgtaagc agaacgaaaa
     2941 cattcgtttc ctggaactgg atgcatacca ccagatccgt aaagctgtag ataaagttta
     3001 cggcaagggt aaatccagca gcttcctgcc gatcgacttt gataccttcg gttacgcgct
     3061 gaccctgttt aaagacgatg cggatatcca ctcttacctg cgcaacgagt acgaacagaa
     3121 cggcaaacgt atgcctaacc tggctaacaa agtttacgat aacgagctga ttgaagaact
     3181 gctgaacctg tccttcacta aattcggtca cctgtctctg aaagctctgc gttccatcct
     3241 gccgtatatg gaacagggtg aagtctactc ctccgcttgt gaacgtgcag gctacacctt
     3301 caccggtccg aaaaagaagc aaaaaactat gctgctgccg aacatccagc cgattgcgaa
     3361 ccctgtagta atgcgtgcac tgacccaggc gcgcaaagta gtcaacgcga tcatcaaaaa
     3421 gtacggcagc ccggtttcca tccatatcga actggcgcgc gacctgagcc agacttttga
     3481 cgagcgtcgt aaaactaaaa aggaacagga tgaaaaccgt aaaaaaaacg aaaccgcgat
     3541 ccgccagctg atggaatacg gtctgactct gaaccctact ggtcacgata ttgtgaagtt
     3601 caagctgtgg tctgaacaga acggtcgctg tgcttactct ctgcagccga tcgagatcga
     3661 acgtctgctg gagccaggtt acgttgaagt agatcatgtg atcccgtact cccgctctct
     3721 ggatgattct tataccaaca aagttctggt tctgactcgc gaaaaccgtg agaaaggcaa
     3781 ccgcatccca gctgaatatc tgggtgttgg cactgagcgt tggcaacagt tcgaaacctt
     3841 cgtcctgacc aataaacagt tctctaaaaa gaaacgtgac cgtctgctgc gtctgcacta
     3901 cgatgaaaac gaagagactg aattcaaaaa ccgtaacctg aacgatactc gctacatcag
     3961 ccgcttcttc gcaaacttca ttcgtgaaca cctgaaattt gcggaatccg acgataaaca
     4021 gaaagtttat accgtaaacg gccgtgttac cgcccacctg cgttctcgct gggagttcaa
     4081 caagaaccgt gaggaaagcg atctgcacca cgctgttgac gccgttattg tggcgtgcac
     4141 caccccaagc gatatcgcta aggtgaccgc attctaccag cgtcgtgagc agaacaagga
     4201 actggccaaa aaaaccgaac cgcattttcc gcagccgtgg ccgcacttcg cggacgaact
     4261 gcgtgctcgt ctgtccaaac atcctaaaga aagcatcaaa gctctgaacc tgggtaacta
     4321 cgatgaccaa aaactggaat ctctgcagcc ggtgtttgtc agccgtatgc cgaaacgttc
     4381 tgttactggc gctgcgcacc gtgaaacgct gcgccgttac gtgggcatcg acgaacgctc
     4441 cggtaaaatc cagaccgtag taaaaaccaa actgtccaag attaaactgg atgcatccgg
     4501 ccacttccca atgtacggta aagaatccga tccacgcact tatgaagcca tccgccagcg
     4561 tctgctggag cataacaacg acccgaaaaa ggcattccag gggcctctgt acaaaccgaa
     4621 aaaaaacggc gaaccgggcc cggtaatccg tactgtaaaa attatcgaca cgagaaacca
     4681 ggtgatccct ctgaacgatg gtaaaaccgt ggcctacaat tccaacatcg ttcgcgtgga
     4741 cgtgttcgaa aaagatggta aatactactg tgtaccggtg tataccatgg acatcatgaa
     4801 aggcattctg ccgaacaaag cgattgaacc gaacaagccg tactctgaat ggaaagaaat
     4861 gaccgaagat tacacgtttc gtttcagcct gtatccgaac gacctgatcc gcatcgaact
     4921 gccgcgtgaa aaaaccgtta aaaccgctgc aggcgaagaa attaacgtga aagacgtgtt
     4981 cgtttactat aaaacgatcg actccgcaaa cggcggcctg gaactgattt ctcacgacca
     5041 ccgtttctct ctgcgtggcg ttggctctcg caccctgaaa cgtttcgaga aatatcaagt
     5101 tgatgttctg ggtaacatct ataaagtgcg tggcgagaaa cgtgtcggtc tggcgtcctc
     5161 cgcacacagc aaacctggca aaaccattcg tccactgcaa tctactcgtg acgggtcacc
     5221 taaaaaaaag agaaaagtag gcggatctcc taagaagaag agaaaagttt aataggtaat
     5281 ctctgcttaa aagcacagaa tctaagatcc ctgccatttg gcggggattt ttttatttgt
     5341 tttcaggaaa taaataatcg atcgcgtaat aaaatctatt attatttttg tgaagaataa
     5401 atttgggtgc aatgagaatg cgcaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     5461 aaaacctctg acacatgtcg agtaaggatc tccaggcatc aaataaaacg aaaggctcag
     5521 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cgtcgacact
     5581 atggttgctt tgacgtatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     5641 catcaggcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5701 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5761 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5821 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5881 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     5941 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6001 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     6061 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6121 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     6181 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6241 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     6301 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     6361 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     6421 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6481 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6541 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6601 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6661 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6721 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6781 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6841 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6901 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6961 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7021 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7081 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7141 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7201 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7261 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7321 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7381 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7441 tgctggcctt ttgctcacat agtcatgccc cgcgcccacc ggaaggagct gactgggttg
     7501 aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta
     7561 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     7621 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct
     7681 tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg
     7741 cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa
     7801 cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc
     7861 accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt
     7921 ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa
     7981 accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt
     8041 gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc
     8101 taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc
     8161 gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa
     8221 cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata
     8281 gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc
     8341 ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg
     8401 agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac
     8461 gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt
     8521 cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg
     8581 gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa
     8641 cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat
     8701 accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatg
//
