LOCUS       sequence-296086-       12415 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For CRISPR-Cas12a genome editing in E. coli. Contains; 
            constituitively expressed Ascpf1 gene, a heat-inducible lambda red 
            recombination system, an arabinose-inducible CRISPR array targeting 
            pMB1..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12415)
  AUTHORS   Jervis AJ, Hanko EKR, Dunstan MS, Robinson CJ, Takano E, Scrutton NS
  TITLE     A plasmid toolset for CRISPR-mediated genome editing and CRISPRi 
            gene regulation in Escherichia coli.
  JOURNAL   Microb Biotechnol. 2021 Mar 12. doi: 10.1111/1751-7915.13780.
  PUBMED    33710766
REFERENCE   2  (bases 1 to 12415)
  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: "Microb 
            Biotechnol. 2021 Mar 12. doi: 10.1111/1751-7915.13780."
COMMENT     Sequence Label: pSIMcpf1
FEATURES             Location/Qualifiers
     source          1..12415
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    4..22
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     CDS             complement(139..819)
                     /codon_start=1
                     /gene="exo"
                     /product="5' to 3' double-stranded DNA exonuclease in the 
                     lambda Red system"
                     /label=Exo
                     /note="creates 3' single-stranded DNA tails"
                     /translation="MTPDIILQRTGIDVRAVEQGDDAWHKLRLGVITASEVHNVIAKPR
                     SGKKWPDMKMSYFHTLLAEVCTGVAPEVNAKALAWGKQYENDARTLFEFTSGVNVTESP
                     IIYRDESMRTACSPDGLCSDGNGLELKCPFTSRDFMKFRLGGFEAIKSAYMAQVQYSMW
                     VTRKNAWYFANYDPRMKREGLHYVVIERDEKYMASFDEIVPEFIEKMDEALAEIGFVFG
                     EQWR"
     CDS             complement(816..1601)
                     /codon_start=1
                     /gene="bet"
                     /product="single-stranded DNA binding recombinase in the 
                     lambda Red system"
                     /label=Beta
                     /note="mediates pairing of a single-stranded DNA with a 
                     complementary target"
                     /translation="MSTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFI
                     ALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESC
                     TCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLA
                     FGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLP
                     LCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA"
     protein_bind    2034..2048
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     promoter        2491..2525
                     /label=BBa_J23114
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.1"
     CDS             2627..6547
                     /codon_start=1
                     /product="CRISPR-associated protein from Acidaminococcus 
                     sp. BV3L6 (Zetsche et al., 2015)"
                     /label=AsCpf1
                     /translation="MTQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARND
                     HYKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRN
                     AIHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFD
                     KFTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHF
                     ENVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVL
                     NLAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLL
                     RNENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTG
                     KITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTL
                     KKQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKA
                     RNYATKKPYSVEKFKLNFQMPTLASGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYK
                     ALSFEPTEKTSEGFDKMYYDYFPDAAKMIPKCSTQLKAVTAHFQTHTTPILLSNNFIEP
                     LEITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSID
                     LSSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKG
                     HHGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLK
                     DQKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFF
                     HVPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQR
                     SLNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAV
                     VVLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQ
                     FTSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYD
                     VKTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIEN
                     HRFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQM
                     RNSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKES
                     KDLKLQNGISNQDWLAYIQELRN"
     regulatory      6267..6276
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(6630..7508)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        7535..7819
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     7742..7761
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             complement(8146..9171)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(8146..9171)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     promoter        complement(9172..9243)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(9356..10069)
                     /codon_start=1
                     /gene="cIts"
                     /product="temperature-sensitive variant of the phage lambda
                     repressor"
                     /label=lambda repressor (ts)
                     /note="thermosensitivity is conferred by the A67T mutation"
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLTKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPKLRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     RBS             10158..10166
                     /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)"
     misc_feature    10181..10199
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     CDS             complement(10716..11666)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      11714..11936
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 cagctgtctc ttatacacat ctccgctgtg ctttcagtgg atttcggata acagaaaggc
       61 cgggaaatac ccagcctcgc tttgtaacgg agtagacgaa agtgattgcg cctacccgga
      121 tattatcgtg aggatgcgtc atcgccattg ctccccaaat acaaaaccaa tttcagccag
      181 tgcctcgtcc attttttcga tgaactccgg cacgatctcg tcaaaactcg ccatgtactt
      241 ttcatcccgc tcaatcacga cataatgcag gccttcacgc ttcatacgcg ggtcatagtt
      301 ggcaaagtac caggcatttt ttcgcgtcac ccacatgctg tactgcacct gggccatgta
      361 agctgacttt atggcctcga aaccaccgag ccggaacttc atgaaatccc gggaggtaaa
      421 cgggcatttc agttcaaggc cgttgccgtc actgcataaa ccatcgggag agcaggcggt
      481 acgcatactt tcgtcgcgat agatgatcgg ggattcagta acattcacgc cggaagtgaa
      541 ctcaaacagg gttctggcgt cgttctcgta ctgttttccc caggccagtg ctttagcgtt
      601 aacttccgga gccacaccgg tgcaaacctc agcaagcagg gtgtggaagt aggacatttt
      661 catgtcaggc cacttctttc cggagcgggg ttttgctatc acgttgtgaa cttctgaagc
      721 ggtgatgacg ccgagccgta atttgtgcca cgcatcatcc ccctgttcga cagctctcac
      781 atcgatcccg gtacgctgca ggataatgtc cggtgtcatg ctgccacctt ctgctctgcg
      841 gctttctgtt tcaggaatcc aagagctttt actgcttcgg cctgtgtcag ttctgacgat
      901 gcacgaatgt cgcggcgaaa tatctgggaa cagagcggca ataagtcgtc atcccatgtt
      961 ttatccaggg cgatcagcag agtgttaatc tcctgcatgg tttcatcgtt aaccggagtg
     1021 atgtcgcgtt ccggctgacg ttctgcagtg tatgcagtat tttcgacaat gcgctcggct
     1081 tcatccttgt catagatacc agcaaatccg aaggccagac gggcacactg aatcatggct
     1141 ttatgacgta acatccgttt gggatgcgac tgccacggcc ccgtgatttc tctgccttcg
     1201 cgagttttga atggttcgcg gcggcattca tccatccatt cggtaacgca gatcggatga
     1261 ttacggtcct tgcggtaaat ccggcatgta caggattcat tgtcctgctc aaagtccatg
     1321 ccatcaaact gctggttttc attgatgatg cgggaccagc catcaacgcc caccaccgga
     1381 acgatgccat tctgcttatc aggaaaggcg taaatttctt tcgtccacgg attaaggccg
     1441 tactggttgg caacgatcag taatgcgatg aactgcgcat cgctggcatc acctttaaat
     1501 gccgtctggc gaagagtggt gatcagttcc tgtgggtcga cagaatccat gccgacacgt
     1561 tcagccagct tcccagccag cgttgcgagt gcagtactca ttcgttttat acctctgaat
     1621 caatatcaac ctggtggtga gcaatggttt caaccatgta ccggatgtgt tctgccatgc
     1681 gctcctgaaa ctcaacatcg tcatcaaacg cacgggtaat ggattttttg ctggccccgt
     1741 ggcgttgcaa atgatcgatg catagcgatt caaacaggtg ctggggcagg cctttttcca
     1801 tgtcgtctgc cagttctgcc tctttctctt cacgggcgag ctgctggtag tgacgcgccc
     1861 agctctgagc ctcaagacga tcctgaatgt aataagcgtt catggctgaa ctcctgaaat
     1921 agctgtgaaa atatcgcccg cgaaatgccg ggctgattag taatccggaa tcgcacttac
     1981 ggccaatgct tcgtttcgta tcacacaccc caaagccttc tgctttgaat gctgcccttc
     2041 ttcagggctt aatttttaag agcgtcacct tcatggtggt cagtgcgtcc tgctgatgtg
     2101 ctcagtatca ccgccagtgg tatttatgtc aacaccgcca gagataattt atcaccgcag
     2161 atggttatct gtatgttttt tatatgaatt tattttttgc aggggggcat tgtttggtag
     2221 gtgagagatc tgaattgcta tgtttagtga gttgtatcta tttatttttc aataaataca
     2281 attggttatg tgttttgggg gcgatcgtga ggcaaagaaa acccggcgct gaggccgggt
     2341 taagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2401 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     2461 ctacggggtc tgacgctcag tggaacgaaa ttgatggcta gctcagtcct aggtacaatg
     2521 ctagcgaatt caaaagatct acaccgtcat aaaatgcctc gcatgtcagt gttattctct
     2581 tgcgcggttc ataatccatg aaactgtttt tatgtaggag gtcgtgatga cccagtttga
     2641 aggttttacc aatctttatc aggtaagcaa gacgctgaga tttgagctga ttccccaggg
     2701 caaaacactg aaacacattc aagagcaagg cttcattgaa gaggataagg cccgaaacga
     2761 ccattataaa gaacttaaac cgattatcga tcgcatctac aagacgtatg cagaccagtg
     2821 tctgcaactg gtgcagttag attgggaaaa tttaagcgcg gctatagact cttaccgtaa
     2881 ggagaaaaca gaagaaactc gtaatgcatt aatcgaggag caggctacat accgcaatgc
     2941 aattcatgac tattttattg gtaggacaga taatctaacg gacgccatca acaagcgtca
     3001 tgccgaaata tacaaaggtc tgtttaaagc ggagctgttc aacggtaaag tattaaagca
     3061 gttaggtacc gtaacgacca cggaacatga aaatgccttg ctgcgttctt ttgataaatt
     3121 cactacgtat ttttccgggt tctacgagaa tcgcaagaac gtgtttagcg cagaagatat
     3181 tagtacggcc attccgcatc gtattgttca agacaatttt ccgaaattta aagaaaattg
     3241 tcatattttt actcgtctga tcacagcagt gccatcactg cgagaacatt ttgaaaatgt
     3301 taagaaggcg attggcattt tcgtgtcaac cagcattgaa gaagtgtttt cattcccgtt
     3361 ctataatcag cttcttaccc agacacagat tgacctatat aatcagctct taggtggtat
     3421 tagccgagaa gctggaacgg aaaagataaa aggcctgaat gaggtgctga acctggcaat
     3481 tcaaaagaac gatgaaacag cgcacattat tgcatctctc ccccatcgat ttataccgct
     3541 ctttaaacaa attctgtccg accgcaacac actgagcttt atcctggagg aatttaaaag
     3601 cgatgaagaa gtaattcaga gcttctgcaa atataaaacg ttactgcgca atgaaaacgt
     3661 cctggaaacg gctgaggccc tctttaacga actgaacagc attgatctca cccatatttt
     3721 tatttcacat aaaaagctgg aaactatcag cagcgcgctg tgcgaccatt gggacacgct
     3781 gagaaatgcc ctgtatgaac gtcgaatttc cgagctgacc ggaaagatca ccaagtcagc
     3841 taaagagaaa gtccagcgct ccctcaaaca tgaagatatt aacttacaag agattatttc
     3901 ggcggccggt aaggaactgt cggaggcttt caaacagaag acaagcgaaa tcctctcaca
     3961 tgcgcacgcg gccctagatc agcctctgcc gacgacgctg aaaaagcagg aagagaaaga
     4021 aatcctgaaa tctcaattag attctctact gggactgtac cacttgctgg actggtttgc
     4081 ggtggacgag tctaatgagg tggaccctga atttagcgca cgcctgaccg gtataaaatt
     4141 ggagatggaa ccgtcgttga gcttttacaa caaagcacgc aactatgcga ccaaaaagcc
     4201 ttattcagtg gaaaaattca aattgaactt ccaaatgccg accctagcgt caggttggga
     4261 tgtcaacaaa gagaagaaca acggtgcgat cttatttgtg aaaaacggcc tgtattatct
     4321 gggcattatg ccgaagcaga aaggccgtta caaggcactg tcattcgaac cgacggagaa
     4381 gaccagtgaa gggtttgata aaatgtatta cgactacttt ccggatgccg ccaaaatgat
     4441 tccgaagtgt tcgacccagc tgaaagcggt gaccgcgcat tttcagacac acaccacccc
     4501 gatcctgctt tcaaataact ttatagaacc gctggaaata accaaagaaa tttacgacct
     4561 gaacaatccg gaaaaagaac cgaagaaatt tcagaccgcc tacgcaaaga aaaccggaga
     4621 tcaaaaaggt taccgcgaag ccctgtgtaa atggattgat tttacgcgcg attttctcag
     4681 caaatacacc aagaccactt caatagattt gagttcgctg cgtccgagct cccagtataa
     4741 agacctgggc gaatattatg ccgagctgaa cccgttactg taccacatta gcttccagcg
     4801 cattgcggaa aaagaaatca tggatgcagt cgaaactgga aagctttatc tgttccagat
     4861 ttacaacaag gattttgcga aaggccatca cggcaaaccg aatctacata ctctctattg
     4921 gactggcctg ttctctcccg aaaacttggc caagacctca attaaactca atggccaagc
     4981 cgagctgttc taccgtccga aatcgcgcat gaagcggatg gcccatcgtc ttggggagaa
     5041 aatgctcaat aagaaactga aggatcagaa aacaccgatc cctgacacgc tgtatcagga
     5101 actttacgac tatgtcaatc accgtctgag tcacgatctg tcggacgaag cacgggcctt
     5161 actgccgaac gtaataacca aagaggtgtc acacgagatc attaaggatc gacgttttac
     5221 ctccgacaaa ttctttttcc acgtaccgat tacccttaat taccaggcgg cgaactcacc
     5281 ctcgaaattt aaccagcgcg taaacgcgta cctgaaagag cacccggaaa cccctattat
     5341 tgggattgac cgtggcgaac gcaatcttat ctatataacc gtaatcgact ctaccggcaa
     5401 gattttagaa cagcgtagcc taaacaccat tcagcaattt gattaccaga aaaagttaga
     5461 taatcgggaa aaagaacgtg tggcggcacg gcaggcttgg tcggtagttg ggacgattaa
     5521 ggatcttaaa caaggttatt tatcccaggt tatacacgaa atcgtagact taatgataca
     5581 ttatcaagcc gttgtggtgc tggaaaacct gaacttcgga tttaagtcaa agcggacggg
     5641 cattgccgaa aaggcggtgt atcagcagtt cgagaagatg ctgatcgata aactcaattg
     5701 cttagtacta aaagattatc ccgctgaaaa ggtaggtggc gttctcaacc cgtaccagct
     5761 tacagatcaa tttaccagct ttgcgaagat gggtacacag agcggttttc tgttttatgt
     5821 tcccgcaccg tataccagca aaatcgatcc cctgaccggc tttgttgatc cttttgtatg
     5881 gaaaaccatc aaaaaccacg aaagccgcaa acatttcctg gagggctttg attttcttca
     5941 ctacgatgtg aaaacaggcg attttattct gcacttcaaa atgaatcgca atcttagctt
     6001 tcagcgcggt ctgcctgggt ttatgccggc gtgggatatt gtgttcgaaa agaatgaaac
     6061 gcagttcgac gccaaaggca caccgtttat tgccgggaaa cgtatagtac cggtgatcga
     6121 aaatcatcgg tttacaggcc gttatcgcga cctgtacccg gctaatgagc tgattgcact
     6181 gctcgaagaa aagggaattg tatttcgtga cggttccaat attctgccta aactgttgga
     6241 aaatgatgac tcgcacgcga ttgataccat ggtggctctt atccgcagcg ttcttcaaat
     6301 gcgtaactct aacgcggcga cgggggaaga ttatattaac agcccggtac gcgatctcaa
     6361 cggcgtctgt tttgatagtc gcttccagaa ccccgaatgg ccgatggacg cggacgccaa
     6421 cggagcatac catattgcgc tgaaaggcca gctgttactg aatcatctca aagaatccaa
     6481 agacctcaaa ctacagaatg gcatttcaaa tcaggattgg ctcgcgtata ttcaggaact
     6541 gcgcaattga tagtccatgt gaccttcacc ctcatggacc gcaaaaaacc ccgcttcggc
     6601 ggggtttttt cgctgcgcaa atcgatttat tatgacaact tgacggctac atcattcact
     6661 ttttcttcac aaccggcacg gaactcgctc gggctggccc cggtgcattt tttaaatacc
     6721 cgcgagaaat agagttgatc gtcaaaacca acattgcgac cgacggtggc gataggcatc
     6781 cgggtggtgc tcaaaagcag cttcgcctgg ctgatacgtt ggtcctcgcg ccagcttaag
     6841 acgctaatcc ctaactgctg gcggaaaaga tgtgacagac gcgacggcga caagcaaaca
     6901 tgctgtgcga cgctggcgat atcaaaattg ctgtctgcca ggtgatcgct gatgtactga
     6961 caagcctcgc gtacccgatt atccatcggt ggatggagcg actcgttaat cgcttccatg
     7021 cgccgcagta acaattgctc aagcagattt atcgccagca gctccgaata gcgcccttcc
     7081 ccttgcccgg cgttaatgat ttgcccaaac aggtcgctga aatgcggctg gtgcgcttca
     7141 tccgggcgaa agaaccccgt attggcaaat attgacggcc agttaagcca ttcatgccag
     7201 taggcgcgcg gacgaaagta aacccactgg tgataccatt cgcgagcctc cggatgacga
     7261 ccgtagtgat gaatctctcc tggcgggaac agcaaaatat cacccggtcg gcaaacaaat
     7321 tctcgtccct gatttttcac caccccctga ccgcgaatgg tgagattgag aatataacct
     7381 ttcattccca gcggtcggtc gataaaaaaa tcgagataac cgttggcctc aatcggcgtt
     7441 aaacccgcca ccagatgggc attaaacgag tatcccggca gcaggggatc attttgcgct
     7501 tcagccatac ttttcatact cccgccattc agagaagaaa ccaattgtcc atattgcatc
     7561 agacattgcc gtcactgcgt cttttactgg ctcttctcgc taaccaaacc ggtaaccccg
     7621 cttattaaaa gcattctgta acaaagcggg accaaagcca tgacaaaaac gcgtaacaaa
     7681 agtgtctata atcacggcag aaaagtccac attgattatt tgcacggcgt cacactttgc
     7741 tatgccatag catttttatc cataagatta gcggatccta cctgacgctt tttatcgcaa
     7801 ctctctactg tttctccatt ttcaatttct actcttgtag attcatagct cacgctgtag
     7861 gtatcaattt ctactcttgt agatccggat caagagctac caactctttt ttttgaagct
     7921 tgggcccgaa caaaaactcg gatccaaact cgagagtata ttttagatga agattatttc
     7981 ttaatctaga aaaaaaaacc ccgcccctga cagggcgggg ttttttttac gttaagggat
     8041 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     8101 ttttaaatca atctaaagta tatatgagta aacttggtct gacagctatt cctttgccct
     8161 cggacgagtg ctggggcgtc ggtttccact atcggcgagt acttctacac agccatcggt
     8221 ccagacggcc gcgcttctgc gggcgatttg tgtacgcccg acagtcccgg ctccggatcg
     8281 gacgattgcg tcgcatcgac cctgcgccca agctgcatca tcgaaattgc cgtcaaccaa
     8341 gctctgatag agttggtcaa gaccaatgcg gagcatatac gcccggagcc gcggcgatcc
     8401 tgcaagctcc ggatgcctcc gctcgaagta gcgcgtctgc tgctccatac aagccaacca
     8461 cggcctccag aagaagatgt tggcgacctc gtattgggaa tccccgaaca tcgcctcgct
     8521 ccagtcaatg accgctgtta tgcggccatt gtccgtcagg acattgttgg agccgaaatc
     8581 cgcgtgcacg aggtgccgga cttcggggca gtcctcggcc caaagcatca gctcatcgag
     8641 agcctgcgcg acggacgcac tgacggtgtc gtccatcaca gtttgccagt gatacacatg
     8701 gggatcagca atcgcgcata tgaaatcacg ccatgtagtg tattgaccga ttccttgcgg
     8761 tccgaatggg ccgaacccgc tcgtctggct aagatcggcc gcagcgatcg catccatggc
     8821 ctccgcgacc ggctgcagaa cagcgggcag ttcggtttca ggcaggtctt gcaacgtgac
     8881 accctgtgca cggcgggaga tgcaataggt caggctctcg ctgaattccc caatgtcaag
     8941 cacttccgga atcgggagcg cggccgatgc aaagtgccga taaacataac gatctttgta
     9001 gaaaccatcg gcgcagctat ttacccgcag gacatatcca cgccctccta catcgaagct
     9061 gaaagcacga gattcttcgc cctccgagag ctgcatcagg tcggagacgc tgtcgaactt
     9121 ttcgatcaga aacttctcga cagacgtcgc ggtgagttca ggctttttca tactcttcct
     9181 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     9241 atgaattcaa tccatttact atgttatgtt ctgaggggag tgaaaattcc cctaattcga
     9301 tgaagattct tgctcaattg ttatcagcta tgcgccgacc agaacacctt gccgatcagc
     9361 caaacgtctc ttcaggccac tgactagcga taactttccc cacaacggaa caactctcat
     9421 tgcatgggat cattgggtac tgtgggttta gtggttgtaa aaacacctga ccgctatccc
     9481 tgatcagttt cttgaaggta aactcatcac ccccaagtct ggctatgcag aaatcacctg
     9541 gctcaacagc ctgctcaggg tcaacgagaa ttaacattcc gtcaggaaag cttggcttgg
     9601 agcctgttgg tgcggtcatg gaattacctt caacctcaag ccagaatgca gaatcactgg
     9661 cttttttggt tgtgcttacc catctctccg catcaccttt ggtaaaggtt ctaagcttag
     9721 gtgagaacat ccctgcctga acatgagaaa aaacagggta ctcatactca cttctaagtg
     9781 acggctgcat actaaccgct tcatacatct cgtagatttc tctggcgatt gaagggctaa
     9841 attcttcaac gctaactttg agaatttttg taagcaatgc ggcgttataa gcatttaatg
     9901 cattgatgcc attaaataaa gcaccaacgc ctgactgccc catccccatc ttgtctgcga
     9961 cagattcctg ggataagcca agttcatttt tctttttttc ataaattgct ttaaggcgac
    10021 gtgcgtcctc aagctgctct tgtgttaatg gtttcttttt tgtgctcata cgttaaatct
    10081 atcaccgcaa gggataaata tctaacaccg tgcgtgttga ctattttacc tctggcggtg
    10141 ataatggttg catgtactaa ggaggttgta tggaacaacg agatgtgtat aagagacagc
    10201 tgacgggttt tgctgcccgc aaacgggctg ttctggtgtt gctagtttgt tatcagaatc
    10261 gcagatccgg cttcaggttt gccggctgaa agcgctattt cttccagaat tgccatgatt
    10321 ttttccccac gggaggcgtc actggctccc gtgttgtcgg cagctttgat tcgataagca
    10381 gcatcgcctg tttcaggctg tctatgtgtg actgttgagc tgtaacaagt tgtctcaggt
    10441 gttcaatttc atgttctagt tgctttgttt tactggtttc acctgttcta ttaggtgtta
    10501 catgctgttc atctgttaca ttgtcgatct gttcatggtg aacagctttg aatgcaccaa
    10561 aaactcgtaa aagctctgat gtatctatct tttttacacc gttttcatct gtgcatatgg
    10621 acagttttcc ctttgatatg taacggtgaa cagttgttct acttttgttt gttagtcttg
    10681 atgcttcact gatagataca agagccataa gaacctcaga tccttccgta tttagccagt
    10741 atgttctcta gtgtggttcg ttgtttttgc gtgagccatg agaacgaacc attgagatca
    10801 tacttacttt gcatgtcact caaaaatttt gcctcaaaac tggtgagctg aatttttgca
    10861 gttaaagcat cgtgtagtgt ttttcttagt ccgttatgta ggtaggaatc tgatgtaatg
    10921 gttgttggta ttttgtcacc attcattttt atctggttgt tctcaagttc ggttacgaga
    10981 tccatttgtc tatctagttc aacttggaaa atcaacgtat cagtcgggcg gcctcgctta
    11041 tcaaccacca atttcatatt gctgtaagtg tttaaatctt tacttattgg tttcaaaacc
    11101 cattggttaa gccttttaaa ctcatggtag ttattttcaa gcattaacat gaacttaaat
    11161 tcatcaaggc taatctctat atttgccttg tgagttttct tttgtgttag ttcttttaat
    11221 aaccactcat aaatcctcat agagtatttg ttttcaaaag acttaacatg ttccagatta
    11281 tattttatga atttttttaa ctggaaaaga taaggcaata tctcttcact aaaaactaat
    11341 tctaattttt cgcttgagaa cttggcatag tttgtccact ggaaaatctc aaagccttta
    11401 accaaaggat tcctgatttc cacagttctc gtcatcagct ctctggttgc tttagctaat
    11461 acaccataag cattttccct actgatgttc atcatctgag cgtattggtt ataagtgaac
    11521 gataccgtcc gttctttcct tgtagggttt tcaatcgtgg ggttgagtag tgccacacag
    11581 cataaaatta gcttggtttc atgctccgtt aagtcatagc gactaatcgc tagttcattt
    11641 gctttgaaaa caactaattc agacatacat ctcaattggt ctaggtgatt ttaatcacta
    11701 taccaattga gatgggctag tcaatgataa ttactagtcc ttttcctttg agttgtgggt
    11761 atctgtaaat tctgctagac ctttgctgga aaacttgtaa attctgctag accctctgta
    11821 aattccgcta gacctttgtg tgtttttttt gtttatattc aagtggttat aatttataga
    11881 ataaagaaag aataaaaaaa gataaaaaga atagatccca gccctgtgta taactcacta
    11941 ctttagtcag ttccgcagta ttacaaaagg atgtcgcaaa cgctgtttgc tcctctacaa
    12001 aacagacctt aaaaccctaa aggcttaagt agcaccctcg caagctcggg caaatcgctg
    12061 aatattcctt ttgtctccga ccatcaggca cctgagtcgc tgtctttttc gtgacattca
    12121 gttcgctgcg ctcacggctc tggcagtgaa tgggggtaaa tggcactaca ggcgcctttt
    12181 atggattcat gcaaggaaac tacccataat acaagaaaag cccgtcacgg gcttctcagg
    12241 gcgttttatg gcgggtctgc tatgtggtgc tatctgactt tttgctgttc agcagttcct
    12301 gccctctgat tttccagtct gaccacttcg gattatcccg tgacaggtca ttcagactgg
    12361 ctaatgcacc cagtaaggca gcggtatcat caacaggctt acccgtctta ctgtc
//
