LOCUS       sequence-494403-       10141 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPRi-mediated Knockdown of RPA0650.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10141)
  AUTHORS   Kathol M, Chowdhury NB, Immethun C, Alsiyabi A, Morris D, Naldrett 
            MJ, Saha R
  TITLE     High enzyme promiscuity in lignin degradation mechanisms in 
            Rhodopseudomonas palustris CGA009.
  JOURNAL   Appl Environ Microbiol. 2025 Aug 20;91(8):e0057325. doi: 
            10.1128/aem.00573-25. Epub 2025 Jul 8.
  PUBMED    40626778
REFERENCE   2  (bases 1 to 10141)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl 
            Environ Microbiol."; date: "2025-08-20"; volume: "91(8):e0057325. 
            doi"; pages: " 10.1128/aem.00573-25. Epub 2025 Jul 8"
COMMENT     Sequence Label: pSSBIO32_dCas9_RPA0650
FEATURES             Location/Qualifiers
     source          1..10141
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..772
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             773..1435
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     CDS             complement(1580..2662)
                     /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     2624..2643
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2663..2740)
                     /gene="lacI"
                     /label=lacI promoter
     protein_bind    2861..2882
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2897..2902,2903..2920,2921..2927)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2935..2951
                     /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)."
     CDS             join(2993..3019,3020..3022,3023..5509,5510..5512,
                     5513..7099)
                     /codon_start=1
                     /product="catalytically dead mutant of the Cas9 
                     endonuclease from the Streptococcus pyogenes Type II 
                     CRISPR/Cas system"
                     /label=dCas9
                     /note="RNA-guided DNA-binding protein that lacks 
                     endonuclease activity due to the D10A mutation in the RuvC 
                     catalytic domain and the H840A mutation in the HNH 
                     catalytic domain"
                     /translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
                     NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
                     FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
                     FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
                     ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
                     IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
                     QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
                     RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
                     NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
                     FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
                     SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
                     LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
                     QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
                     HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
                     LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
                     SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
                     VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
                     NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
                     EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
                     ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
                     TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
                     LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
                     ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
                     DATLIHQSITGLYETRIDLSQLGGD"
     terminator      7167..7238
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7254..7281
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(7309..7326)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(7437..7466)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     promoter        7520..7554
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     misc_RNA        7578..7653
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      7754..7785
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(7834..8628)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7945..7964)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     8555..8574
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gcggccaccg gctggctcgc ttcgctcggc ccgtggacaa ccctgctgga caagctgatg
       61 gacaggctgc gcctgcccac gagcttgacc acagggattg cccaccggct acccagcctt
      121 cgaccacata cccaccggct ccaactgcgc ggcctgcggc cttgccccat caattttttt
      181 aattttctct ggggaaaagc ctccggcctg cggcctgcgc gcttcgcttg ccggttggac
      241 accaagtgga aggcgggtca aggctcgcgc agcgaccgcg cagcggcttg gccttgacgc
      301 gcctggaacg acccaagcct atgcgagtgg gggcagtcga agggcgaagc ccgcccgcct
      361 gccccccgag cctcacggcg gcgagtgcgg gggttccaag ggggcagcgc caccttgggc
      421 aaggccgaag gccgcgcagt cgatcaacaa gccccggagg ggccactttt tgccggaggg
      481 ggagccgcgc cgaaggcgtg ggggaacccc gcaggggtgc ccttctttgg gcaccaaaga
      541 actagatata gggcgaaatg cgaaagactt aaaaatcaac aacttaaaaa aggggggtac
      601 gcaacagctc attgcggcac cccccgcaat agctcattgc gtaggttaaa gaaaatctgt
      661 aattgactgc cacttttacg caacgcataa ttgttgtcgc gctgccgaaa agttgcagct
      721 gattgcgcat ggtgccgcaa ccgtgcggca cccctaccgc atggagataa gcatggccac
      781 gcagtccaga gaaatcggca ttcaagccaa gaacaagccc ggtcactggg tgcaaacgga
      841 acgcaaagcg catgaggcgt gggccgggct tattgcgagg aaacccacgg cggcaatgct
      901 gctgcatcac ctcgtggcgc agatgggcca ccagaacgcc gtggtggtca gccagaagac
      961 actttccaag ctcatcggac gttctttgcg gacggtccaa tacgcagtca aggacttggt
     1021 ggccgagcgc tggatctccg tcgtgaagct caacggcccc ggcaccgtgt cggcctacgt
     1081 ggtcaatgac cgcgtggcgt ggggccagcc ccgcgaccag ttgcgcctgt cggtgttcag
     1141 tgccgccgtg gtggttgatc acgacgacca ggacgaatcg ctgttggggc atggcgacct
     1201 gcgccgcatc ccgaccctgt atccgggcga gcagcaacta ccgaccggcc ccggcgagga
     1261 gccgcccagc cagcccggca ttccgggcat ggaaccagac ctgccagcct tgaccgaaac
     1321 ggaggaatgg gaacggcgcg ggcagcagcg cctgccgatg cccgatgagc cgtgttttct
     1381 ggacgatggc gagccgttgg agccgccgac acgggtcacg ctgccgcgcc ggtagcactt
     1441 gggttgcgca gcaacccgta agtgcgctgt tccagactat cggctgtagc cgcctcgccg
     1501 ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga
     1561 cctgaatgga agcacgtcct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     1621 gcattaatga atcggccaat gcgcggggag aggcggtttg cgtattgggc gccagggtgg
     1681 tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag
     1741 agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg
     1801 tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga
     1861 tgtccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct
     1921 gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt
     1981 gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat
     2041 tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg
     2101 ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc
     2161 gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa
     2221 gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca
     2281 gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt
     2341 tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat
     2401 cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg
     2461 tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa
     2521 tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc
     2581 tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat
     2641 cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc
     2701 atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc
     2761 ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcacgacg
     2821 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     2881 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     2941 gcggataaca atttcaaaag aattcaaaag atctaaagag gacttcggat ctatggataa
     3001 gaaatactca ataggcttag ctatcggcac aaatagcgtc ggatgggcgg tgatcactga
     3061 tgaatataag gttccgtcta aaaagttcaa ggttctggga aatacagacc gccacagtat
     3121 caaaaaaaat cttatagggg ctcttttatt tgacagtgga gagacagcgg aagcgactcg
     3181 tctcaaacgg acagctcgta gaaggtatac acgtcggaag aatcgtattt gttatctaca
     3241 ggagattttt tcaaatgaga tggcgaaagt agatgatagt ttctttcatc gacttgaaga
     3301 gtcttttttg gtggaagaag acaagaagca tgaacgtcat cctatttttg gaaatatagt
     3361 agatgaagtt gcttatcatg agaaatatcc aactatctat catctgcgaa aaaaattggt
     3421 agattctact gataaagcgg atttgcgctt aatctatttg gccttagcgc atatgattaa
     3481 gtttcgtggt cattttttga ttgagggaga tttaaatcct gataatagtg atgtggacaa
     3541 actatttatc cagttggtac aaacctacaa tcaattattt gaagaaaacc ctattaacgc
     3601 aagtggagta gatgctaaag cgattctttc tgcacgattg agtaaatcaa gacgattaga
     3661 aaatctcatt gctcagctcc ccggtgagaa gaaaaatggc ttatttggga atctcattgc
     3721 tttgtcattg ggtttgaccc ctaattttaa atcaaatttt gatttggcag aagatgctaa
     3781 attacagctt tcaaaagata cttacgatga tgatttagat aatttattgg cgcaaattgg
     3841 agatcaatat gctgatttgt ttttggcagc taagaattta tcagatgcta ttttactttc
     3901 agatatccta agagtaaata ctgaaataac taaggctccc ctatcagctt caatgattaa
     3961 acgctacgat gaacatcatc aagacttgac tcttttaaaa gctttagttc gacaacaact
     4021 tccagaaaag tataaagaaa tcttttttga tcaatcaaaa aacggatatg caggttatat
     4081 tgatggggga gctagccaag aagaatttta taaatttatc aaaccaattt tagaaaaaat
     4141 ggatggtact gaggaattat tggtgaaact aaatcgtgaa gatttgctgc gcaagcaacg
     4201 gacctttgac aacggctcta ttccccatca aattcacttg ggtgagctgc atgctatttt
     4261 gagaagacaa gaagactttt atccattttt aaaagacaat cgtgagaaga ttgaaaaaat
     4321 cttgactttt cgaattcctt attatgttgg tccattggcg cgtggcaata gtcgttttgc
     4381 atggatgact cggaagtctg aagaaacaat taccccatgg aattttgaag aagttgtcga
     4441 taaaggtgct tcagctcaat catttattga acgcatgaca aactttgata aaaatcttcc
     4501 aaatgaaaaa gtactaccaa aacatagttt gctttatgag tattttacgg tttataacga
     4561 attgacaaag gtcaaatatg ttactgaagg aatgcgaaaa ccagcatttc tttcaggtga
     4621 acagaagaaa gccattgttg atttactctt caaaacaaat cgaaaagtaa ccgttaagca
     4681 attaaaagaa gattatttca aaaaaataga atgttttgat agtgttgaaa tttcaggagt
     4741 tgaagataga tttaatgctt cattaggtac ctaccatgat ttgctaaaaa ttattaaaga
     4801 taaagatttt ttggataatg aagaaaatga agatatctta gaggatattg ttttaacatt
     4861 gaccttattt gaagataggg agatgattga ggaaagactt aaaacatatg ctcacctctt
     4921 tgatgataag gtgatgaaac agcttaaacg tcgccgttat actggttggg gacgtttgtc
     4981 tcgaaaattg attaatggta ttagggataa gcaatctggc aaaacaatat tagatttttt
     5041 gaaatcagat ggttttgcca atcgcaattt tatgcagctg atccatgatg atagtttgac
     5101 atttaaagaa gacattcaaa aagcacaagt gtctggacaa ggcgatagtt tacatgaaca
     5161 tattgcaaat ttagctggta gccctgctat taaaaaaggt attttacaga ctgtaaaagt
     5221 tgttgatgaa ttggtcaaag taatggggcg gcataagcca gaaaatatcg ttattgaaat
     5281 ggcacgtgaa aatcagacaa ctcaaaaggg ccagaaaaat tcgcgagagc gtatgaaacg
     5341 aatcgaagaa ggtatcaaag aattaggaag tcagattctt aaagagcatc ctgttgaaaa
     5401 tactcaattg caaaatgaaa agctctatct ctattatctc caaaatggaa gagacatgta
     5461 tgtggaccaa gaattagata ttaatcgttt aagtgattat gatgtcgatg ccattgttcc
     5521 acaaagtttc cttaaagacg attcaataga caataaggtc ttaacgcgtt ctgataaaaa
     5581 tcgtggtaaa tcggataacg ttccaagtga agaagtagtc aaaaagatga aaaactattg
     5641 gagacaactt ctaaacgcca agttaatcac tcaacgtaag tttgataatt taacgaaagc
     5701 tgaacgtgga ggtttgagtg aacttgataa agctggtttt atcaaacgcc aattggttga
     5761 aactcgccaa atcactaagc atgtggcaca aattttggat agtcgcatga atactaaata
     5821 cgatgaaaat gataaactta ttcgagaggt taaagtgatt accttaaaat ctaaattagt
     5881 ttctgacttc cgaaaagatt tccaattcta taaagtacgt gagattaaca attaccatca
     5941 tgcccatgat gcgtatctaa atgccgtcgt tggaactgct ttgattaaga aatatccaaa
     6001 acttgaatcg gagtttgtct atggtgatta taaagtttat gatgttcgta aaatgattgc
     6061 taagtctgag caagaaatag gcaaagcaac cgcaaaatat ttcttttact ctaatatcat
     6121 gaacttcttc aaaacagaaa ttacacttgc aaatggagag attcgcaaac gccctctaat
     6181 cgaaactaat ggggaaactg gagaaattgt ctgggataaa gggcgagatt ttgccacagt
     6241 gcgcaaagta ttgtccatgc cccaagtcaa tattgtcaag aaaacagaag tacagacagg
     6301 cggattctcc aaggagtcaa ttttaccaaa aagaaattcg gacaagctta ttgctcgtaa
     6361 aaaagactgg gatccaaaaa aatatggtgg ttttgatagt ccaacggtag cttattcagt
     6421 cctagtggtt gctaaggtgg aaaaagggaa atcgaagaag ttaaaatccg ttaaagagtt
     6481 actagggatc acaattatgg aaagaagttc ctttgaaaaa aatccgattg actttttaga
     6541 agctaaagga tataaggaag ttaaaaaaga cttaatcatt aaactaccta aatatagtct
     6601 ttttgagtta gaaaacggtc gtaaacggat gctggctagt gccggagaat tacaaaaagg
     6661 aaatgagctg gctctgccaa gcaaatatgt gaatttttta tatttagcta gtcattatga
     6721 aaagttgaag ggtagtccag aagataacga acaaaaacaa ttgtttgtgg agcagcataa
     6781 gcattattta gatgagatta ttgagcaaat cagtgaattt tctaagcgtg ttattttagc
     6841 agatgccaat ttagataaag ttcttagtgc atataacaaa catagagaca aaccaatacg
     6901 tgaacaagca gaaaatatta ttcatttatt tacgttgacg aatcttggag ctcccgctgc
     6961 ttttaaatat tttgatacaa caattgatcg taaacgatat acgtctacaa aagaagtttt
     7021 agatgccact cttatccatc aatccatcac tggtctttat gaaacacgca ttgatttgag
     7081 tcagctagga ggtgactgaa gtatatttta gatgaagacg aaaactacgc tctggctgct
     7141 taactcgagt aaggatctcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     7201 cctttcgttt tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac
     7261 cttcgggtgg gcctttctgc gtttatacct agggatatat tccgcttcct cgctcactga
     7321 ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg gcttacgaac ggggcggaga
     7381 tttcctggaa gatgccagac gtctaagaaa aggaatattc agcaatttgc ccgtgccgaa
     7441 gaaaggccca cccgtgaagg tgagccagtg agttgattgc tacgtaatta gttagttagc
     7501 ccttagtgac tcgaattcgt tgacagctag ctcagtccta ggtataatgc tagccctatc
     7561 ctcaccgaaa cccaggggtt ttagagctag aaatagcaag ttaaaataag gctagtccgt
     7621 tatcaacttg aaaaagtggc accgagtcgg tgcttttttt ctagataatt ggagaccgag
     7681 ctggtctctc taggatgtgc actagtagag ctctgcagga gtcactaagg gttagttagt
     7741 tagattagca gaaagtcaaa agcctccgac cggaggcttt tgactaaaac ttcccttggg
     7801 cttggtcggt catttcgaac cccagagtcc cgctcagaag aactcgtcaa gaaggcgata
     7861 gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga agcggtcagc
     7921 ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt cctgatagcg
     7981 gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat tttccaccat
     8041 gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt cgggcatgcg
     8101 cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt cgtccagatc
     8161 atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc gatgtttcgc
     8221 ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca ttgcatcagc
     8281 catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct gccccggcac
     8341 ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca cagctgcgca
     8401 aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcctgca gttcattcag
     8461 ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg acagccggaa
     8521 cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga atagcctctc
     8581 cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc gaaacgatcc
     8641 tcatcctgtc tcttgatcag atcttgatcc cctgcgccat cagatccttg gcggcaagaa
     8701 agccatccag tttactttgc agggcttccc aaccttacca gagggcgccc cagctggcaa
     8761 ttccggttcg cttgctgtcc ataaaaccgc ccagtctagc tatcgccatg taagcccact
     8821 gcaagctacc tgctttctct ttgcgcttgc gttttccctt gtccagatag ccatgcatgc
     8881 ttgcgttttc ccttgtccag atagcccagt agctgacatt catcccaggt ggcacttttc
     8941 ggggaaatgt gcgcgcccgc gttcctgctg gcgctgggcc tgtttctggc gctggacttc
     9001 ccgctgttcc gtcagcagct tttcgcccac ggccttgatg atcgcggcgg ccttggcctg
     9061 catatcccga ttcaacggcc ccagggcgtc cagaacgggc ttcaggcgct cccgaaggtc
     9121 tcgggccgtc tcttgggctt gatcggcctt cttgcgcatc tcacgcgctc ctgcggcggc
     9181 ctgtagggca ggctcatacc cctgccgaac cgcttttgtc agccggtcgg ccacggcttc
     9241 cggcgtctca acgcgctttg agattcccag cttttcggcc aatccatgcg gtgcataggc
     9301 gcgtggctcg accgcttgcg ggctgatggt gacgtggccc actggtggcc gctccagggc
     9361 ctcgtagaac gcctgaatgc gcgtgtgacg tgccttgctg ccctcgatgc cccgttgcag
     9421 ccctagatcg gccacagcgg ccgcaaacgt ggtctggtcg cgggtcatct gcgctttgtt
     9481 gccgatgaac tccttggccg acagcctgcc gtcctgcgtc agcggcacca cgaacgcggt
     9541 catgtgcggg ctggtttcgt cacggtggat gctggccgtc acgatgcgat ccgccccgta
     9601 cttgtccgcc agccacttgt gcgccttctc gaagaacgcc gcctgctgtt cttggctggc
     9661 cgacttccac cattccgggc tggccgtcat gacgtactcg accgccaaca cagcgtcctt
     9721 gcgccgcttc tctggcagca actcgcgcag tcggcccatc gcttcatcgg tgctgctggc
     9781 cgcccagtgc tcgttctctg gcgtcctgct ggcgtcagcg ttgggcgtct cgcgctcgcg
     9841 gtaggcgtgc ttgagactgg ccgccacgtt gcccattttc gccagcttct tgcatcgcat
     9901 gatcgcgtat gccgccatgc ctgcccctcc cttttggtgt ccaaccggct cgacgggggc
     9961 agcgcaaggc ggtgcctccg gcgggccact caatgcttga gtatactcac tagactttgc
    10021 ttcgcaaagt cgtgaccgcc tacggcggct gcggcgccct acgggcttgc tctccgggct
    10081 tcgccctgcg cggtcgctgc gctcccttgc cagcccgtgg atatgtggac gatggccgcg
    10141 a
//
