LOCUS       sequence-320039-       12418 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 12418)
  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 12418)
  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..12418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    2..20
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     CDS             complement(137..817)
                     /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="MTPDIILQRTGIDVRAVEQGEDAWHKLRLGVITASEVHNVIAKPR
                     SGKKWPDMKMSYFHTLLAEVCTGVAPEVNAKALAWGKQYENDARTLFEFTSGVNVTESP
                     IIYRDESMRTACSPDGLCSDGNGLELKCPFTSRDFMKFRLGGFEAIKSAYMAQVQYSMW
                     VTRKNAWYFANYDPRMKREGLHYVVIERDEKYMASFDEIVPEFIEKMDEALAEIGFVFG
                     EQWR"
     CDS             complement(814..1599)
                     /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    2032..2046
                     /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        2489..2523
                     /label=BBa_J23114
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.1"
     CDS             2625..6545
                     /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      6265..6274
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(6628..7506)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        7533..7817
                     /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     7740..7759
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             complement(8144..9169)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             complement(8144..9169)
                     /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(9170..9241)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(9354..10067)
                     /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
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     RBS             10156..10164
                     /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    10179..10197
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase 
                     recognition (Reznikoff et al., 2004)"
     CDS             complement(10717..11667)
                     /codon_start=1
                     /gene="rep101 encoding an A56V mutation"
                     /product="temperature-sensitive version of the RepA protein
                     needed for replication with the pSC101 origin (Armstrong et
                     al., 1984)"
                     /label=Rep101(Ts)
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      11715..11937
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
ORIGIN
        1 gctgtctctt atacacatct ccgctgtgct ttcagtggat ttcggataac agaaaggccg
       61 ggaaataccc agcctcgctt tgtaacggag tagacgaaag tgattgcgcc tacccggata
      121 ttatcgtgag gatgcgtcat cgccattgct ccccaaatac aaaaccaatt tcagccagtg
      181 cctcgtccat tttttcgatg aactccggca cgatctcgtc aaaactcgcc atgtactttt
      241 catcccgctc aatcacgaca taatgcaggc cttcacgctt catacgcggg tcatagttgg
      301 caaagtacca ggcatttttt cgcgtcaccc acatgctgta ctgcacctgg gccatgtaag
      361 ctgactttat ggcctcgaaa ccaccgagcc ggaacttcat gaaatcccgg gaggtaaacg
      421 ggcatttcag ttcaaggccg ttgccgtcac tgcataaacc atcgggagag caggcggtac
      481 gcatactttc gtcgcgatag atgatcgggg attcagtaac attcacgccg gaagtaaatt
      541 caaacagggt tctggcgtcg ttctcgtact gttttcccca ggccagtgct ttagcgttaa
      601 cttccggagc cacaccggtg caaacctcag caagcagggt gtggaagtag gacattttca
      661 tgtcaggcca cttctttccg gagcggggtt ttgctatcac gttgtgaact tctgaagcgg
      721 tgatgacgcc gagccgtaat ttgtgccacg catcttcccc ctgttcgaca gctctcacat
      781 cgatcccggt acgctgcagg ataatgtccg gtgtcatgct gccaccttct gctctgcggc
      841 tttctgtttc aggaatccaa gagcttttac tgcttcggcc tgtgtcagtt ctgacgatgc
      901 acgaatgtcg cggcgaaata tctgggaaca gagcggcaat aagtcgtcat cccatgtttt
      961 atccagggcg atcagcagag tgttaatctc ctgcatggtt tcatcgttaa ccggagtgat
     1021 gtcgcgttcc ggctgacgtt ctgcagtgta tgcagtattt tcgacaatgc gctcggcttc
     1081 atccttgtca tagataccag caaatccgaa ggccagacgg gcacactgaa tcatggcttt
     1141 atgacgtaac atccgtttgg gatgcgactg ccacggcccc gtgatttctc tgccttcgcg
     1201 agttttgaat ggttcgcggc ggcattcatc catccattcg gtaacgcaga tcggatgatt
     1261 acggtccttg cggtaaatcc ggcatgtaca ggattcattg tcctgctcaa agtccatgcc
     1321 atcaaactgc tggttttcat tgatgatgcg ggaccagcca tcaacgccca ccaccggaac
     1381 gatgccattc tgcttatcag gaaaggcgta aatttctttc gtccacggat taaggccgta
     1441 ctggttggca acgatcagta atgcgatgaa ctgcgcatcg ctggcatcac ctttaaatgc
     1501 cgtctggcga agagtggtga tcagttcctg tgggtcgaca gaatccatgc cgacacgttc
     1561 agccagcttc ccagccagcg ttgcgagtgc agtactcatt cgttttatac ctctgaatca
     1621 atatcaacct ggtggtgagc aatggtttca accatgtacc ggatgtgttc tgccatgcgc
     1681 tcctgaaact caacatcgtc atcaaacgca cgggtaatgg attttttgct ggccccgtgg
     1741 cgttgcaaat gatcgatgca tagcgattca aacaggtgct ggggcaggcc tttttccatg
     1801 tcgtctgcca gttctgcctc tttctcttca cgggcgagct gctggtagtg acgcgcccag
     1861 ctctgagcct caagacgatc ctgaatgtaa taagcgttca tggctgaact cctgaaatag
     1921 ctgtgaaaat atcgcccgcg aaatgccggg ctgattagta atccggaatc gcacttacgg
     1981 ccaatgcttc gtttcgtatc acacacccca aagccttctg ctttgaatgc tgcccttctt
     2041 cagggcttaa tttttaagag cgtcaccttc atggtggtca gtgcgtcctg ctgatgtgct
     2101 cagtatcacc gccagtggta tttatgtcaa caccgccaga gataatttat caccgcagat
     2161 ggttatctgt atgtttttta tatgaattta ttttttgcag gggggcattg tttggtaggt
     2221 gagagatctg aattgctatg tttagtgagt tgtatctatt tatttttcaa taaatacaat
     2281 tggttatgtg ttttgggggc gatcgtgagg caaagaaaac ccggcgctga ggccgggtta
     2341 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     2401 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     2461 acggggtctg acgctcagtg gaacgaaatt gatggctagc tcagtcctag gtacaatgct
     2521 agcgaattca aaagatctac accgtcataa aatgcctcgc atgtcagtgt tattctcttg
     2581 cgcggttcat aatccatgaa actgttttta tgtaggaggt cgtgatgacc cagtttgaag
     2641 gttttaccaa tctttatcag gtaagcaaga cgctgagatt tgagctgatt ccccagggca
     2701 aaacactgaa acacattcaa gagcaaggct tcattgaaga ggataaggcc cgaaacgacc
     2761 attataaaga acttaaaccg attatcgatc gcatctacaa gacgtatgca gaccagtgtc
     2821 tgcaactggt gcagttagat tgggaaaatt taagcgcggc tatagactct taccgtaagg
     2881 agaaaacaga agaaactcgt aatgcattaa tcgaggagca ggctacatac cgcaatgcaa
     2941 ttcatgacta ttttattggt aggacagata atctaacgga cgccatcaac aagcgtcatg
     3001 ccgaaatata caaaggtctg tttaaagcgg agctgttcaa cggtaaagta ttaaagcagt
     3061 taggtaccgt aacgaccacg gaacatgaaa atgccttgct gcgttctttt gataaattca
     3121 ctacgtattt ttccgggttc tacgagaatc gcaagaacgt gtttagcgca gaagatatta
     3181 gtacggccat tccgcatcgt attgttcaag acaattttcc gaaatttaaa gaaaattgtc
     3241 atatttttac tcgtctgatc acagcagtgc catcactgcg agaacatttt gaaaatgtta
     3301 agaaggcgat tggcattttc gtgtcaacca gcattgaaga agtgttttca ttcccgttct
     3361 ataatcagct tcttacccag acacagattg acctatataa tcagctctta ggtggtatta
     3421 gccgagaagc tggaacggaa aagataaaag gcctgaatga ggtgctgaac ctggcaattc
     3481 aaaagaacga tgaaacagcg cacattattg catctctccc ccatcgattt ataccgctct
     3541 ttaaacaaat tctgtccgac cgcaacacac tgagctttat cctggaggaa tttaaaagcg
     3601 atgaagaagt aattcagagc ttctgcaaat ataaaacgtt actgcgcaat gaaaacgtcc
     3661 tggaaacggc tgaggccctc tttaacgaac tgaacagcat tgatctcacc catattttta
     3721 tttcacataa aaagctggaa actatcagca gcgcgctgtg cgaccattgg gacacgctga
     3781 gaaatgccct gtatgaacgt cgaatttccg agctgaccgg aaagatcacc aagtcagcta
     3841 aagagaaagt ccagcgctcc ctcaaacatg aagatattaa cttacaagag attatttcgg
     3901 cggccggtaa ggaactgtcg gaggctttca aacagaagac aagcgaaatc ctctcacatg
     3961 cgcacgcggc cctagatcag cctctgccga cgacgctgaa aaagcaggaa gagaaagaaa
     4021 tcctgaaatc tcaattagat tctctactgg gactgtacca cttgctggac tggtttgcgg
     4081 tggacgagtc taatgaggtg gaccctgaat ttagcgcacg cctgaccggt ataaaattgg
     4141 agatggaacc gtcgttgagc ttttacaaca aagcacgcaa ctatgcgacc aaaaagcctt
     4201 attcagtgga aaaattcaaa ttgaacttcc aaatgccgac cctagcgtca ggttgggatg
     4261 tcaacaaaga gaagaacaac ggtgcgatct tatttgtgaa aaacggcctg tattatctgg
     4321 gcattatgcc gaagcagaaa ggccgttaca aggcactgtc attcgaaccg acggagaaga
     4381 ccagtgaagg gtttgataaa atgtattacg actactttcc ggatgccgcc aaaatgattc
     4441 cgaagtgttc gacccagctg aaagcggtga ccgcgcattt tcagacacac accaccccga
     4501 tcctgctttc aaataacttt atagaaccgc tggaaataac caaagaaatt tacgacctga
     4561 acaatccgga aaaagaaccg aagaaatttc agaccgccta cgcaaagaaa accggagatc
     4621 aaaaaggtta ccgcgaagcc ctgtgtaaat ggattgattt tacgcgcgat tttctcagca
     4681 aatacaccaa gaccacttca atagatttga gttcgctgcg tccgagctcc cagtataaag
     4741 acctgggcga atattatgcc gagctgaacc cgttactgta ccacattagc ttccagcgca
     4801 ttgcggaaaa agaaatcatg gatgcagtcg aaactggaaa gctttatctg ttccagattt
     4861 acaacaagga ttttgcgaaa ggccatcacg gcaaaccgaa tctacatact ctctattgga
     4921 ctggcctgtt ctctcccgaa aacttggcca agacctcaat taaactcaat ggccaagccg
     4981 agctgttcta ccgtccgaaa tcgcgcatga agcggatggc ccatcgtctt ggggagaaaa
     5041 tgctcaataa gaaactgaag gatcagaaaa caccgatccc tgacacgctg tatcaggaac
     5101 tttacgacta tgtcaatcac cgtctgagtc acgatctgtc ggacgaagca cgggccttac
     5161 tgccgaacgt aataaccaaa gaggtgtcac acgagatcat taaggatcga cgttttacct
     5221 ccgacaaatt ctttttccac gtaccgatta cccttaatta ccaggcggcg aactcaccct
     5281 cgaaatttaa ccagcgcgta aacgcgtacc tgaaagagca cccggaaacc cctattattg
     5341 ggattgaccg tggcgaacgc aatcttatct atataaccgt aatcgactct accggcaaga
     5401 ttttagaaca gcgtagccta aacaccattc agcaatttga ttaccagaaa aagttagata
     5461 atcgggaaaa agaacgtgtg gcggcacggc aggcttggtc ggtagttggg acgattaagg
     5521 atcttaaaca aggttattta tcccaggtta tacacgaaat cgtagactta atgatacatt
     5581 atcaagccgt tgtggtgctg gaaaacctga acttcggatt taagtcaaag cggacgggca
     5641 ttgccgaaaa ggcggtgtat cagcagttcg agaagatgct gatcgataaa ctcaattgct
     5701 tagtactaaa agattatccc gctgaaaagg taggtggcgt tctcaacccg taccagctta
     5761 cagatcaatt taccagcttt gcgaagatgg gtacacagag cggttttctg ttttatgttc
     5821 ccgcaccgta taccagcaaa atcgatcccc tgaccggctt tgttgatcct tttgtatgga
     5881 aaaccatcaa aaaccacgaa agccgcaaac atttcctgga gggctttgat tttcttcact
     5941 acgatgtgaa aacaggcgat tttattctgc acttcaaaat gaatcgcaat cttagctttc
     6001 agcgcggtct gcctgggttt atgccggcgt gggatattgt gttcgaaaag aatgaaacgc
     6061 agttcgacgc caaaggcaca ccgtttattg ccgggaaacg tatagtaccg gtgatcgaaa
     6121 atcatcggtt tacaggccgt tatcgcgacc tgtacccggc taatgagctg attgcactgc
     6181 tcgaagaaaa gggaattgta tttcgtgacg gttccaatat tctgcctaaa ctgttggaaa
     6241 atgatgactc gcacgcgatt gataccatgg tggctcttat ccgcagcgtt cttcaaatgc
     6301 gtaactctaa cgcggcgacg ggggaagatt atattaacag cccggtacgc gatctcaacg
     6361 gcgtctgttt tgatagtcgc ttccagaacc ccgaatggcc gatggacgcg gacgccaacg
     6421 gagcatacca tattgcgctg aaaggccagc tgttactgaa tcatctcaaa gaatccaaag
     6481 acctcaaact acagaatggc atttcaaatc aggattggct cgcgtatatt caggaactgc
     6541 gcaattgata gtccatgtga ccttcaccct catggaccgc aaaaaacccc gcttcggcgg
     6601 ggttttttcg ctgcgcaaat cgatttatta tgacaacttg acggctacat cattcacttt
     6661 ttcttcacaa ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg
     6721 cgagaaatag agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg
     6781 ggtggtgctc aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac
     6841 gctaatccct aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg
     6901 ctgtgcgacg ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca
     6961 agcctcgcgt acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg
     7021 ccgcagtaac aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc
     7081 ttgcccggcg ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc
     7141 cgggcgaaag aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta
     7201 ggcgcgcgga cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc
     7261 gtagtgatga atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc
     7321 tcgtccctga tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt
     7381 cattcccagc ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa
     7441 acccgccacc agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc
     7501 agccatactt ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag
     7561 acattgccgt cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct
     7621 tattaaaagc attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag
     7681 tgtctataat cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta
     7741 tgccatagca tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact
     7801 ctctactgtt tctccatttt caatttctac tcttgtagat tcatagctca cgctgtaggt
     7861 atcaatttct actcttgtag atccggatca agagctacca actctttttt ttgaagcttg
     7921 ggcccgaaca aaaactcgga tccaaactcg agagtatatt ttagatgaag attatttctt
     7981 aatctagaaa aaaaaacccc gcccctgaca gggcggggtt ttttttacgt taagggattt
     8041 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     8101 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagctattcc tttgccctcg
     8161 gacgagtgct ggggcgtcgg tttccactat cggcgagtac ttctacacag ccatcggtcc
     8221 agacggccgc gcttctgcgg gcgatttgtg tacgcccgac agtcccggct ccggatcgga
     8281 cgattgcgtc gcatcgaccc tgcgcccaag ctgcatcatc gaaattgccg tcaaccaagc
     8341 tctgatagag ttggtcaaga ccaatgcgga gcatatacgc ccggagccgc ggcgatcctg
     8401 caagctccgg atgcctccgc tcgaagtagc gcgtctgctg ctccatacaa gccaaccacg
     8461 gcctccagaa gaagatgttg gcgacctcgt attgggaatc cccgaacatc gcctcgctcc
     8521 agtcaatgac cgctgttatg cggccattgt ccgtcaggac attgttggag ccgaaatccg
     8581 cgtgcacgag gtgccggact tcggggcagt cctcggccca aagcatcagc tcatcgagag
     8641 cctgcgcgac ggacgcactg acggtgtcgt ccatcacagt ttgccagtga tacacatggg
     8701 gatcagcaat cgcgcatatg aaatcacgcc atgtagtgta ttgaccgatt ccttgcggtc
     8761 cgaatgggcc gaacccgctc gtctggctaa gatcggccgc agcgatcgca tccatggcct
     8821 ccgcgaccgg ctgcagaaca gcgggcagtt cggtttcagg caggtcttgc aacgtgacac
     8881 cctgtgcacg gcgggagatg caataggtca ggctctcgct gaattcccca atgtcaagca
     8941 cttccggaat cgggagcgcg gccgatgcaa agtgccgata aacataacga tctttgtaga
     9001 aaccatcggc gcagctattt acccgcagga catatccacg ccctcctaca tcgaagctga
     9061 aagcacgaga ttcttcgccc tccgagagct gcatcaggtc ggagacgctg tcgaactttt
     9121 cgatcagaaa cttctcgaca gacgtcgcgg tgagttcagg ctttttcata ctcttccttt
     9181 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     9241 gaattcaatc catttactat gttatgttct gaggggagtg aaaattcccc taattcgatg
     9301 aagattcttg ctcaattgtt atcagctatg cgccgaccag aacaccttgc cgatcagcca
     9361 aacgtctctt caggccactg actagcgata actttcccca caacggaaca actctcattg
     9421 catgggatca ttgggtactg tgggtttagt ggttgtaaaa acacctgacc gctatccctg
     9481 atcagtttct tgaaggtaaa ctcatcaccc ccaagtctgg ctatgcagaa atcacctggc
     9541 tcaacagcct gctcagggtc aacgagaatt aacattccgt caggaaagct tggcttggag
     9601 cctgttggtg cggtcatgga attaccttca acctcaagcc agaatgcaga atcactggct
     9661 tttttggttg tgcttaccca tctctccgca tcacctttgg taaaggttct aagctcaggt
     9721 gagaacatcc ctgcctgaac atgagaaaaa acagggtact catactcact tctaagtgac
     9781 ggctgcatac taaccgcttc atacatctcg tagatttctc tggcgattga agggctaaat
     9841 tcttcaacgc taactttgag aatttttgta agcaatgcgg cgttataagc atttaatgca
     9901 ttgatgccat taaataaagc accaacgcct gactgcccca tccccatctt gtctgcgaca
     9961 gattcctggg ataagccaag ttcatttttc tttttttcat aaattgcttt aaggcgacgt
    10021 gcgtcctcaa gctgctcttg tgttaatggt ttcttttttg tgctcatacg ttaaatctat
    10081 caccgcaagg gataaatatc taacaccgtg cgtgttgact attttacctc tggcggtgat
    10141 aatggttgca tgtactaagg aggttgtatg gaacaacgag atgtgtataa gagacagctg
    10201 acgggttttg ctgcccgcaa acgggctgtt ctggtgttgc tagtttgtta tcagaatcgc
    10261 agatccggct tcagccggtt tgccggctga aagcgctatt tcttccagaa ttgccatgat
    10321 tttttcccca cgggaggcgt cactggctcc cgtgttgtcg gcagctttga ttcgataagc
    10381 agcatcgcct gtttcaggct gtctatgtgt gactgttgag ctgtaacaag ttgtctcagg
    10441 tgttcaattt catgttctag ttgctttgtt ttactggttt cacctgttct attaggtgtt
    10501 acatgctgtt catctgttac attgtcgatc tgttcatggt gaacagcttt gaatgcacca
    10561 aaaactcgta aaagctctga tgtatctatc ttttttacac cgttttcatc tgtgcatatg
    10621 gacagttttc cctttgatat gtaacggtga acagttgttc tacttttgtt tgttagtctt
    10681 gatgcttcac tgatagatac aagagccata agaacctcag atccttccgt atttagccag
    10741 tatgttctct agtgtggttc gttgtttttg cgtgagccat gagaacgaac cattgagatc
    10801 atacttactt tgcatgtcac tcaaaaattt tgcctcaaaa ctggtgagct gaatttttgc
    10861 agttaaagca tcgtgtagtg tttttcttag tccgttatgt aggtaggaat ctgatgtaat
    10921 ggttgttggt attttgtcac cattcatttt tatctggttg ttctcaagtt cggttacgag
    10981 atccatttgt ctatctagtt caacttggaa aatcaacgta tcagtcgggc ggcctcgctt
    11041 atcaaccacc aatttcatat tgctgtaagt gtttaaatct ttacttattg gtttcaaaac
    11101 ccattggtta agccttttaa actcatggta gttattttca agcattaaca tgaacttaaa
    11161 ttcatcaagg ctaatctcta tatttgcctt gtgagttttc ttttgtgtta gttcttttaa
    11221 taaccactca taaatcctca tagagtattt gttttcaaaa gacttaacat gttccagatt
    11281 atattttatg aattttttta actggaaaag ataaggcaat atctcttcac taaaaactaa
    11341 ttctaatttt tcgcttgaga acttggcata gtttgtccac tggaaaatct caaagccttt
    11401 aaccaaagga ttcctgattt ccacagttct cgtcatcagc tctctggttg ctttagctaa
    11461 tacaccataa gcattttccc tactgatgtt catcatctga acgtattggt tataagtgaa
    11521 cgataccgtc cgttctttcc ttgtagggtt ttcaatcgtg gggttgagta gtgccacaca
    11581 gcataaaatt agcttggttt catgctccgt taagtcatag cgactaatcg ctagttcatt
    11641 tgctttgaaa acaactaatt cagacataca tctcaattgg tctaggtgat tttaatcact
    11701 ataccaattg agatgggcta gtcaatgata attactagtc cttttccttt gagttgtggg
    11761 tatctgtaaa ttctgctaga cctttgctgg aaaacttgta aattctgcta gaccctctgt
    11821 aaattccgct agacctttgt gtgttttttt tgtttatatt caagtggtta taatttatag
    11881 aataaagaaa gaataaaaaa agataaaaag aatagatccc agccctgtgt ataactcact
    11941 actttagtca gttccgcagt attacaaaag gatgtcgcaa acgctgtttg ctcctctaca
    12001 aaacagacct taaaacccta aaggcttaag tagcaccctc gcaagctcgg gcaaatcgct
    12061 gaatattcct tttgtctccg accatcaggc acctgagtcg ctgtcttttt cgtgacattc
    12121 agttcgctgc gctcacggct ctggcagtga atgggggtaa atggcactac aggcgccttt
    12181 tatggattca tgcaaggaaa ctacccataa tacaagaaaa gcccgtcacg ggcttctcag
    12241 ggcgttttat ggcgggtctg ctatgtggtg ctatctgact ttttgctgtt cagcagttcc
    12301 tgccctctga ttttccagtc tgaccacttc ggattatccc gtgacaggtc attcagactg
    12361 gctaatgcac ccagtaaggc agcggtatca tcaacaggct tacccgtctt actgtcca
//
