LOCUS       sequence-298594-        9496 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Shuttle vector for expression dCas9, mobilization cluster, expresses
            the tracrRNA and a CRISPR array designed for the easy cloning of new
            spacers..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9496)
  AUTHORS   Tan SI, Yu PJ, Ng IS
  TITLE     CRISPRi-mediated Programming Essential Gene can as a Direct 
  JOURNAL   Biotechnol Bioeng. 2020 May 27. doi: 10.1002/bit.27443.
  PUBMED    32458463
REFERENCE   2  (bases 1 to 9496)
  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: "Biotechnol 
            Bioeng. 2020 May 27. doi: 10.1002/bit.27443."
COMMENT     Sequence Label: pSM-dCas9::Ap
FEATURES             Location/Qualifiers
     source          1..9496
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            214..323
                     /note="incP origin of transfer"
     CDS             356..727
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     terminator      1665..1722
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(2041..2058)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2175..2720)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(2255..2274)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2832..2837,2838..2854,2855..2860)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     misc_RNA        complement(3178..3256)
                     /label=tracrRNA
                     /note="trans-activating CRISPR RNA for the Streptococcus 
                     pyogenes CRISPR/Cas9 system"
     CDS             join(3552..3578,3579..3581,3582..6068,6069..6071,
                     6072..7658)
                     /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"
     misc_feature    7679..7810
                     /label=crRNA leader
                     /note="crRNA leader sequence for the Streptococcus pyogenes
                     CRISPR/Cas system"
     repeat_region   7811..7846
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     repeat_region   7877..7912
                     /label=DR
                     /note="direct repeat for the Streptococcus pyogenes 
                     CRISPR/Cas system"
     primer_bind     complement(8341..8360)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        8566..8629
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8630..8698,8699..9490)
                     /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(8848..8867)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gtcccctcag ttcagtaatt tcctgcattt gcctgtttcc agtcggtaga tattccacaa
       61 aacagcaggg aagcagcgct tttccgctgc ataaccctgc ttcggggtca ttatagcgat
      121 tttttcggta tatccatcct ttttcgcacg atatacagga ttttgccaaa gggttcgtgt
      181 agactttcct tggtgtatcc aacggcgtca gccgggcagg ataggtgaag taggcccacc
      241 cgcgagcggg tgttccttct tcactgtccc ttattcgcac ctggcggtgc tcaacgggaa
      301 tcctgctctg cgaggctggc cggctaccgc cggcgtaaca gatgagggca agcggatggc
      361 tgatgaaacc aagccaacca ggaagggcag cccacctatc aaggtgtact gccttccaga
      421 cgaacgaaga gcgattgagg aaaaggcggc ggcggccggc atgagcctgt cggcctacct
      481 gctggccgtc ggccagggct acaaaatcac gggcgtcgtg gactatgagc acgtccgcga
      541 gctggcccgc atcaatggcg acctgggccg cctgggcggc ctgctgaaac tctggctcac
      601 cgacgacccg cgcacggcgc ggttcggtga tgccacgatc ctcgccctgc tggcgaagat
      661 cgaagagaag caggacgagc ttggcaaggt catgatgggc gtggtccgcc cgagggcaga
      721 gccatgactt ttttagccgc taaaacggcc ggggggtgcg cgtgattgcc aagcacgtcc
      781 ccatgcgctc catcaagaag agcgacttcg cggagctggt gaagtacatc accgacgagc
      841 agggcaagac cgagcgcctg ggtcacgtgc gcgtcacgaa ctgcgaggca aacaccctgc
      901 ccgctgtcat tggcgaggtg atggcgacca gcacggcaac acccgtttcc gaggccgaca
      961 agacctatca cctgctggtt agcttccgcg cgggagagaa gcccgacgcg gagacgttgc
     1021 gcgcgattga ggaccgcatc tgcgctgggc ttggcttcgc cgagcatcag cgcgtcagtg
     1081 ccgtgcatca cgacaccgac aacctgcaca tccatatcgc catcaacaag attcacccga
     1141 cccgaaacac catccatgag ccgtatcggg cctaccgcgc cctcgctgac ctctgcgcga
     1201 cgctcgaacg ggactacggg cttgagcgtg acaatcacga aacgcggcag cgcgtttccg
     1261 agaaccgcgc gaacgacatg gagcggcacg cgggcgtgga aagcctggtc ggctggatcc
     1321 ggccacgatg cgtccggcgt agaggatctg aagatcagca gttcaacctg ttgatagtac
     1381 gtactaagct ccatgtttca cgtactaagc tctcatgttt aacgtactaa gctctcatgt
     1441 ttaacgaact aaaccctcat ggctaacgta ctaagctctc atggctaacg tactaagctc
     1501 tcatgtttca cgtactaagc tctcatgttt gaacaataaa attaatataa atcagcaact
     1561 taaatagcct ctaaggtttt aagttttata agaaaaaaaa gaatatataa ggcttttaaa
     1621 gcttttaagg tttaacggtt gtggactagt agcggccgct gcagtccggc aaaaaagggc
     1681 aaggtgtcac caccctgccc tttttcttta aaaccgaaaa gattacttcg cgttatgcag
     1741 atcttattta ttcggcgcaa agtgcgtcgg gtgatgctgc caacttactg atttagtgta
     1801 tgatggtgtt tttgaggtgc tccagtggct tctgtttcta tcagctgtcc ctcctgttca
     1861 gctactgacg gggtggtgcg taacggcaaa agcaccgccg gacatcagcg ctagcggagt
     1921 gtatactggc ttactatgtt ggcactgatg agggtgtcag tgaagtgctt catgtggcag
     1981 gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga tacaggatat attccgcttc
     2041 ctcgctcact gactcgctac gctcggtcgt tcgactgcgg cgagcggaaa tggcttacga
     2101 acggggcgga gatttcctgg aagatgccag gaagatactt aacagggaag tgagagggcc
     2161 gcggcaaagc cgtttttcca taggctccgc ccccctgaca agcatcacga aatctgacgc
     2221 tcaaatcagt ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctggc
     2281 ggctccctcg tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat tccgctgtta
     2341 tggccgcgtt tgtctcattc cacgcctgac actcagttcc gggtaggcag ttcgctccaa
     2401 gctggactgt atgcacgaac cccccgttca gtccgaccgc tgcgccttat ccggtaacta
     2461 tcgtcttgag tccaacccgg aaagacatgc aaaagcacca ctggcagcag ccactggtaa
     2521 ttgatttaga ggagttagtc ttgaagtcat gcgccggtta aggctaaact gaaaggacaa
     2581 gttttggtga ctgcgctcct ccaagccagt tacctcggtt caaagagttg gtagctcaga
     2641 gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa gagattacgc
     2701 gcagaccaaa acgatctcaa gaagatcatc ttattaatca gataaaatat ttctagattt
     2761 cagtgcaatt tatctcttca aatgtagcac ctgaagtcag ccccatacga tataagttgt
     2821 aattctcatg tttgacagct tatcatcgat aagctttaat gcggtagttt atcacagtta
     2881 aattgctaac gcagtcaggc accgtgtatg aaatctaaca atgcgctcat cgtcatcctc
     2941 ggcaccgtca ccctggatgc tgtaggcata ggcttggtta tgccggtact gccgggcctc
     3001 ttgcgggatt acgaaatcat cctgtggagc ttagtaggtt tagcaagatg gcagcgccta
     3061 aatgtagaat gataaaagga ttaagagatt aatttcccta aaaatgataa aacaagcgtt
     3121 ttgaaagcgc ttgttttttt ggtttgcagt cagagtagaa tagaagtatc aaaaaaagca
     3181 ccgactcggt gccacttttt caagttgata acggactagc cttattttaa cttgctatgc
     3241 tgttttgaat ggttccaaca agattatttt ataactttta taacaaataa tcaaggagaa
     3301 attcaaagaa atttatcagc cataaaacaa tacttaatac tatagaatga taacaaaata
     3361 aactactttt taaaagaatt ttgtgttata atctatttat tattaagtat tgggtaatat
     3421 tttttgaaga gatattttga aaaagaaaaa ttaaagcata ttaaactaat ttcggaggtc
     3481 attaaaacta ttattgaaat catcaaactc attatggatt taatttaaac tttttatttt
     3541 aggaggcaaa aatggataag aaatactcaa taggcttagc tatcggcaca aatagcgtcg
     3601 gatgggcggt gatcactgat gaatataagg ttccgtctaa aaagttcaag gttctgggaa
     3661 atacagaccg ccacagtatc aaaaaaaatc ttataggggc tcttttattt gacagtggag
     3721 agacagcgga agcgactcgt ctcaaacgga cagctcgtag aaggtataca cgtcggaaga
     3781 atcgtatttg ttatctacag gagatttttt caaatgagat ggcgaaagta gatgatagtt
     3841 tctttcatcg acttgaagag tcttttttgg tggaagaaga caagaagcat gaacgtcatc
     3901 ctatttttgg aaatatagta gatgaagttg cttatcatga gaaatatcca actatctatc
     3961 atctgcgaaa aaaattggta gattctactg ataaagcgga tttgcgctta atctatttgg
     4021 ccttagcgca tatgattaag tttcgtggtc attttttgat tgagggagat ttaaatcctg
     4081 ataatagtga tgtggacaaa ctatttatcc agttggtaca aacctacaat caattatttg
     4141 aagaaaaccc tattaacgca agtggagtag atgctaaagc gattctttct gcacgattga
     4201 gtaaatcaag acgattagaa aatctcattg ctcagctccc cggtgagaag aaaaatggct
     4261 tatttgggaa tctcattgct ttgtcattgg gtttgacccc taattttaaa tcaaattttg
     4321 atttggcaga agatgctaaa ttacagcttt caaaagatac ttacgatgat gatttagata
     4381 atttattggc gcaaattgga gatcaatatg ctgatttgtt tttggcagct aagaatttat
     4441 cagatgctat tttactttca gatatcctaa gagtaaatac tgaaataact aaggctcccc
     4501 tatcagcttc aatgattaaa cgctacgatg aacatcatca agacttgact cttttaaaag
     4561 ctttagttcg acaacaactt ccagaaaagt ataaagaaat cttttttgat caatcaaaaa
     4621 acggatatgc aggttatatt gatgggggag ctagccaaga agaattttat aaatttatca
     4681 aaccaatttt agaaaaaatg gatggtactg aggaattatt ggtgaaacta aatcgtgaag
     4741 atttgctgcg caagcaacgg acctttgaca acggctctat tccccatcaa attcacttgg
     4801 gtgagctgca tgctattttg agaagacaag aagactttta tccattttta aaagacaatc
     4861 gtgagaagat tgaaaaaatc ttgacttttc gaattcctta ttatgttggt ccattggcgc
     4921 gtggcaatag tcgttttgca tggatgactc ggaagtctga agaaacaatt accccatgga
     4981 attttgaaga agttgtcgat aaaggtgctt cagctcaatc atttattgaa cgcatgacaa
     5041 actttgataa aaatcttcca aatgaaaaag tactaccaaa acatagtttg ctttatgagt
     5101 attttacggt ttataacgaa ttgacaaagg tcaaatatgt tactgaagga atgcgaaaac
     5161 cagcatttct ttcaggtgaa cagaagaaag ccattgttga tttactcttc aaaacaaatc
     5221 gaaaagtaac cgttaagcaa ttaaaagaag attatttcaa aaaaatagaa tgttttgata
     5281 gtgttgaaat ttcaggagtt gaagatagat ttaatgcttc attaggtacc taccatgatt
     5341 tgctaaaaat tattaaagat aaagattttt tggataatga agaaaatgaa gatatcttag
     5401 aggatattgt tttaacattg accttatttg aagataggga gatgattgag gaaagactta
     5461 aaacatatgc tcacctcttt gatgataagg tgatgaaaca gcttaaacgt cgccgttata
     5521 ctggttgggg acgtttgtct cgaaaattga ttaatggtat tagggataag caatctggca
     5581 aaacaatatt agattttttg aaatcagatg gttttgccaa tcgcaatttt atgcagctga
     5641 tccatgatga tagtttgaca tttaaagaag acattcaaaa agcacaagtg tctggacaag
     5701 gcgatagttt acatgaacat attgcaaatt tagctggtag ccctgctatt aaaaaaggta
     5761 ttttacagac tgtaaaagtt gttgatgaat tggtcaaagt aatggggcgg cataagccag
     5821 aaaatatcgt tattgaaatg gcacgtgaaa atcagacaac tcaaaagggc cagaaaaatt
     5881 cgcgagagcg tatgaaacga atcgaagaag gtatcaaaga attaggaagt cagattctta
     5941 aagagcatcc tgttgaaaat actcaattgc aaaatgaaaa gctctatctc tattatctcc
     6001 aaaatggaag agacatgtat gtggaccaag aattagatat taatcgttta agtgattatg
     6061 atgtcgatgc cattgttcca caaagtttcc ttaaagacga ttcaatagac aataaggtct
     6121 taacgcgttc tgataaaaat cgtggtaaat cggataacgt tccaagtgaa gaagtagtca
     6181 aaaagatgaa aaactattgg agacaacttc taaacgccaa gttaatcact caacgtaagt
     6241 ttgataattt aacgaaagct gaacgtggag gtttgagtga acttgataaa gctggtttta
     6301 tcaaacgcca attggttgaa actcgccaaa tcactaagca tgtggcacaa attttggata
     6361 gtcgcatgaa tactaaatac gatgaaaatg ataaacttat tcgagaggtt aaagtgatta
     6421 ccttaaaatc taaattagtt tctgacttcc gaaaagattt ccaattctat aaagtacgtg
     6481 agattaacaa ttaccatcat gcccatgatg cgtatctaaa tgccgtcgtt ggaactgctt
     6541 tgattaagaa atatccaaaa cttgaatcgg agtttgtcta tggtgattat aaagtttatg
     6601 atgttcgtaa aatgattgct aagtctgagc aagaaatagg caaagcaacc gcaaaatatt
     6661 tcttttactc taatatcatg aacttcttca aaacagaaat tacacttgca aatggagaga
     6721 ttcgcaaacg ccctctaatc gaaactaatg gggaaactgg agaaattgtc tgggataaag
     6781 ggcgagattt tgccacagtg cgcaaagtat tgtccatgcc ccaagtcaat attgtcaaga
     6841 aaacagaagt acagacaggc ggattctcca aggagtcaat tttaccaaaa agaaattcgg
     6901 acaagcttat tgctcgtaaa aaagactggg atccaaaaaa atatggtggt tttgatagtc
     6961 caacggtagc ttattcagtc ctagtggttg ctaaggtgga aaaagggaaa tcgaagaagt
     7021 taaaatccgt taaagagtta ctagggatca caattatgga aagaagttcc tttgaaaaaa
     7081 atccgattga ctttttagaa gctaaaggat ataaggaagt taaaaaagac ttaatcatta
     7141 aactacctaa atatagtctt tttgagttag aaaacggtcg taaacggatg ctggctagtg
     7201 ccggagaatt acaaaaagga aatgagctgg ctctgccaag caaatatgtg aattttttat
     7261 atttagctag tcattatgaa aagttgaagg gtagtccaga agataacgaa caaaaacaat
     7321 tgtttgtgga gcagcataag cattatttag atgagattat tgagcaaatc agtgaatttt
     7381 ctaagcgtgt tattttagca gatgccaatt tagataaagt tcttagtgca tataacaaac
     7441 atagagacaa accaatacgt gaacaagcag aaaatattat tcatttattt acgttgacga
     7501 atcttggagc tcccgctgct tttaaatatt ttgatacaac aattgatcgt aaacgatata
     7561 cgtctacaaa agaagtttta gatgccactc ttatccatca atccatcact ggtctttatg
     7621 aaacacgcat tgatttgagt cagctaggag gtgactgaag tatattttag atgaagatta
     7681 tttcttaata actaaaaata tggtataata ctcttaataa atgcagtaat acaggggctt
     7741 ttcaagactg aagtctagct gagacaaata gtgcgattac gaaatttttt agacaaaaat
     7801 agtctacgag gttttagagc tatgctgttt tgaatggtcc caaaactgag accagtctcg
     7861 gaagctcaaa ggtctcgttt tagagctatg ctgttttgaa tggtcccaaa acttcagcac
     7921 actgagactt gttgagttcc atgttttaga gctatgctgt tttgaatgga ctccattcaa
     7981 cattgccgat gataacttga gaaagagggt taataccagc agtcggatac cttcctattc
     8041 tttctgttaa agcgttttca tgttataata ggcaaaagaa gagtagtgtg atcgtccatt
     8101 ccgacagcat cgccagtcac tatggcgtgc tgctagcgct atatgcgttg atgcaatttc
     8161 tatgcgcacc cgttctcgga gcactgtccg accgctttgg ccgccgccca gtcctgctcg
     8221 cttcgctact tggagccact atcgactacg cgatcatggc gaccacaccc gtcctgtgga
     8281 tcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg
     8341 cctatatcgc cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg
     8401 cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct
     8461 ccttgcatgc accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct
     8521 gcttcctaat gcaggagtcg cataagggag agcgtcgact tcaaatatgt atccgctcat
     8581 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     8641 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     8701 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcgc gagtgggtta
     8761 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     8821 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     8881 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     8941 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     9001 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     9061 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     9121 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     9181 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     9241 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     9301 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtcac gcggtatcat
     9361 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     9421 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     9481 gcattggtaa ctcgag
//
