LOCUS       sequence-336407-       10163 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of cas12f1, arabinose-inducible expression 
            of lambda recombinase, and IPTG-inducible expression of sgRNA for 
            curing of pTarget12f1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10163)
  AUTHORS   Okano K, Sato Y, Hizume T, Honda K
  TITLE     Genome editing by miniature CRISPR/Cas12f1 enzyme in Escherichia 
            coli.
  JOURNAL   J Biosci Bioeng. 2021 Aug;132(2):120-124. doi: 
            10.1016/j.jbiosc.2021.04.009. Epub 2021 May 19.
  PUBMED    34023220
REFERENCE   2  (bases 1 to 10163)
  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"; doi: 
            "10.1016/j.jbiosc.2021.04.009"; journalName: "J Biosci Bioeng"; 
            date: "2021-08"; volume: "132"; issue: "2"; pages: "120-124"
COMMENT     Sequence Label: pCas12f1
FEATURES             Location/Qualifiers
     source          1..10163
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    245..261
                     /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             complement(530..1612)
                     /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     1574..1593
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1613..1690)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             complement(1823..2773)
                     /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      2821..3043
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             complement(3695..4510)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3695..4510)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4418..4437
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4502..4521
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     misc_RNA        complement(4666..4744)
                     /label=tracrRNA
                     /note="trans-activating CRISPR RNA for the Streptococcus 
                     pyogenes CRISPR/Cas9 system"
     CDS             complement(6771..7649)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRH
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        7676..7960
                     /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     7883..7902
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             7987..7995
                     /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)"
     CDS             8004..8420
                     /codon_start=1
                     /gene="gam"
                     /product="inhibitor of the host RecBCD nuclease in the 
                     lambda Red system"
                     /label=Gam
                     /note="protects the double-stranded DNA substrate for 
                     recombination"
                     /translation="MDINTETEIKQKHSLTPFPVFLISPAFRGRYFHSYFRSSAMNAYY
                     IQDRLEAQSWARHYQQLAREEKEAELADDMEKGLPQHLFESLCIDHLQRHGASKKSITR
                     AFDDDVEFQERMAEHIRYMVETIAHHQVDIDSEV"
     CDS             8426..9211
                     /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"
     CDS             9208..9888
                     /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"
     terminator      9889..10133
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
ORIGIN
        1 gatctcaaaa aaacgccccc ctgacgagca tcagttgcat tccttcattc gtctattcgg
       61 gttctgcaac tttcttgtgc agaattggag aggaaaaatg aatttgtttc gagggttact
      121 ttccgaagaa agcacttctc gacattaggc tgatgcaagc agcccacctt cactcaagtt
      181 ctaatcccct aagggacagc ttttggtgaa gcggttctcc actttatcag tgaagtgtgt
      241 gaaattgtta tccgctcaca attccacaca acatacgagc cggatgatta attgtcaaca
      301 gctcatttca gaatatttgc cagaaccgtt atgatgtcgg cgcaaaaaac attatccaga
      361 acgggagtgc gccttgagcg acacgaatta tgcagtgatt tacgacctgc acagccatac
      421 cacagcttcc gatggctgcc tgacgccaga agcattggtg caccgtgcag tcgatgataa
      481 gctgtcaaac cagatcaatt cgcgctaact tacattaatt gcgttgcgct cactgcccgc
      541 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
      601 aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca
      661 gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt
      721 gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt
      781 cttcggtatc gtcgtatccc actaccgaga tatccgcacc aacgcgcagc ccggactcgg
      841 taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa
      901 cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc
      961 cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca
     1021 gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac
     1081 ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac
     1141 tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag
     1201 cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc
     1261 gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca
     1321 tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt
     1381 gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc
     1441 ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca
     1501 ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct
     1561 gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca
     1621 ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcaccatt
     1681 cgatggtgtc aacgtaaatg catgccgctc gccttccatg ggtatggaca gttttccctt
     1741 tgatatgtaa cggtgaacag ttgttctact tttgtttgtt agtcttgatg cttcactgat
     1801 agatacaaga gccataagaa cctcagatcc ttccgtattt agccagtatg ttctctagtg
     1861 tggttcgttg tttttgcgtg agccatgaga acgaaccatt gagatcatac ttactttgca
     1921 tgtcactcaa aaattttgcc tcaaaactgg tgagctgaat ttttgcagtt aaagcatcgt
     1981 gtagtgtttt tcttagtccg ttatgtaggt aggaatctga tgtaatggtt gttggtattt
     2041 tgtcaccatt catttttatc tggttgttct caagttcggt tacgagatcc atttgtctat
     2101 ctagttcaac ttggaaaatc aacgtatcag tcgggcggcc tcgcttatca accaccaatt
     2161 tcatattgct gtaagtgttt aaatctttac ttattggttt caaaacccat tggttaagcc
     2221 ttttaaactc atggtagtta ttttcaagca ttaacatgaa cttaaattca tcaaggctaa
     2281 tctctatatt tgccttgtga gttttctttt gtgttagttc ttttaataac cactcataaa
     2341 tcctcataga gtatttgttt tcaaaagact taacatgttc cagattatat tttatgaatt
     2401 tttttaactg gaaaagataa ggcaatatct cttcactaaa aactaattct aatttttcgc
     2461 ttgagaactt ggcatagttt gtccactgga aaatctcaaa gcctttaacc aaaggattcc
     2521 tgatttccac agttctcgtc atcagctctc tggttgcttt agctaataca ccataagcat
     2581 tttccctact gatgttcatc atctgaacgt attggttata agtgaacgat accgtccgtt
     2641 ctttccttgt agggttttca atcgtggggt tgagtagtgc cacacagcat aaaattagct
     2701 tggtttcatg ctccgttaag tcatagcgac taatcgctag ttcatttgct ttgaaaacaa
     2761 ctaattcaga catacatctc aattggtcta ggtgatttta atcactatac caattgagat
     2821 gggctagtca atgataatta ctagtccttt tcctttgagt tgtgggtatc tgtaaattct
     2881 gctagacctt tgctggaaaa cttgtaaatt ctgctagacc ctctgtaaat tccgctagac
     2941 ctttgtgtgt tttttttgtt tatattcaag tggttataat ttatagaata aagaaagaat
     3001 aaaaaaagat aaaaagaata gatcccagcc ctgtgtataa ctcactactt tagtcagttc
     3061 cgcagtatta caaaaggatg tcgcaaacgc tgtttgctcc tctacaaaac agaccttaaa
     3121 accctaaagg cttaagtagc accctcgcaa gctcggttgc ggccgcaatc gggcaaatcg
     3181 ctgaatattc cttttgtctc cgaccatcag gcacctgagt cgctgtcttt ttcgtgacat
     3241 tcagttcgct gcgctcacgg ctctggcagt gaatgggggt aaatggcact acaggcgcct
     3301 tttatggatt catgcaagga aactacccat aatacaagaa aagcccgtca cgggcttctc
     3361 agggcgtttt atggcgggtc tgctatgtgg tgctatctga ctttttgctg ttcagcagtt
     3421 cctgccctct gattttccag tctgaccact tcggattatc ccgtgacagg tcattcagac
     3481 tggctaatgc acccagtaag gcagcggtat catcaacggg gtctgacgct cagtggaacg
     3541 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     3601 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     3661 acagttacgt ttccaaccaa ttaaccaatt ctgattagaa aaactcatcg agcatcaaat
     3721 gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa agccgtttct
     3781 gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt
     3841 ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa
     3901 ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagct
     3961 tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac
     4021 tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat
     4081 cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca
     4141 gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt
     4201 tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga
     4261 tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat
     4321 cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat
     4381 acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat
     4441 ataaatcagc atccatgttg gaatttaatc gcggcctcga gcaagacgtt tcccgttgaa
     4501 tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg
     4561 atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacaca acgtggcttt
     4621 ccctgcaggg tttgcagtca gagtagaata gaagtatcaa aaaaagcacc gactcggtgc
     4681 cactttttca agttgataac ggactagcct tattttaact tgctatgctg ttttgaatgg
     4741 ttccaacaag attattttat aacttttata acaaataatc aaggagaaat tcaaagaaat
     4801 ttatcagcca taaaacaata cttaatacta tagaatgata acaaaataaa ctacttttta
     4861 aaagaatttt gtgttataat ctatttatta ttaagtattg ggtaatattt tttgaagaga
     4921 tattttgaaa aagaaaaatt aaagcatatt aaactaattt cggaggtcat taaaactatt
     4981 attgaaatca tcaaactcat tatggattta atttaaactt tttattttag gaggcaaaag
     5041 tcgagatggc caaaaacacc attaccaaaa cactgaaact gcgtattgtg cgtccgtata
     5101 atagcgcaga agtggaaaaa attgttgccg acgaaaaaaa caaccgcgaa aaaatcgcac
     5161 tggaaaagaa caaagacaaa gtgaaagaag cctgcagcaa acatctgaaa gttgcagcat
     5221 attgtaccac acaggttgaa cgtaatgcat gtctgttttg taaagcccgt aaactggatg
     5281 acaagttcta tcaaaaactg cgtggtcagt ttccggatgc agttttttgg caagaaatca
     5341 gcgaaatttt tcgccagctg cagaaacagg cagcagaaat ctataatcag agcctgatcg
     5401 aactgtacta cgagattttt atcaaaggca aaggtattgc aaatgccagc agcgttgaac
     5461 attatctgag tgatgtttgt tatacccgtg cagcagaact gtttaaaaac gcagcaattg
     5521 caagcggtct gcgtagcaaa atcaaaagca attttcgtct gaaagaactg aaaaacatga
     5581 aaagtggtct gccgaccacc aaaagcgata attttccgat tccgctggtt aaacagaaag
     5641 gtggtcagta taccggtttt gaaattagca atcataatag cgacttcatc atcaagattc
     5701 cgtttggtcg ttggcaggtc aaaaaagaga ttgataaata tcgtccgtgg gagaaatttg
     5761 actttgaaca ggttcagaaa agcccgaaac cgattagcct gctgctgagc acccagcgtc
     5821 gtaaacgtaa taaaggttgg agcaaagatg aaggcaccga agccgaaatc aaaaaagtta
     5881 tgaatggcga ttatcagacc agctacattg aagttaaacg tggcagcaaa atcggtgaaa
     5941 aaagcgcatg gatgctgaat ctgagcattg atgttccgaa aattgataaa ggtgtggatc
     6001 cgagcattat tggtggtatt gatgttggtg ttaaatcacc gctggtttgc gcaattaaca
     6061 atgcatttag ccgttatagc atcagcgata acgacctgtt tcacttcaac aagaaaatgt
     6121 ttgcacgtcg tcgtatcctg ctgaaaaaaa accgtcataa acgtgcaggt catggtgcaa
     6181 aaaacaaact gaaaccgatc accattctga ccgaaaaaag tgaacgtttt cgcaaaaagc
     6241 tgattgaacg ttgggcatgt gaaatcgcgg atttcttcat taaaaacaaa gttggcaccg
     6301 tgcagatgga aaatctggaa agcatgaaac gtaaagagga cagctatttt aacattcgcc
     6361 tgcgtggctt ttggccgtat gcagaaatgc agaacaaaat cgagtttaag ctgaaacagt
     6421 atggcatcga aattcgtaaa gtggcaccga ataataccag caaaacctgt agcaaatgtg
     6481 gccatctgaa caactatttc aacttcgagt accgcaagaa aaacaaattc ccgcacttta
     6541 aatgcgaaaa atgcaacttc aaagaaaacg ccgattataa tgcagccctg aatatttcaa
     6601 acccgaaact gaaaagcacc aaagaggaac cgtaaggtac cagtatattt tagatgaaga
     6661 ttatttctta atctagacat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6721 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg catcgattta ttatgacaac
     6781 ttgacggcta catcattcac tttttcttca caaccggcac ggaactcgct cgggctggcc
     6841 ccggtgcatt ttttaaatac ccgcgagaag tagagttgat cgtcaaaacc aacattgcga
     6901 ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt
     6961 tggtcctcgc gccagcttaa gacgctaatc cctaactgct ggcggaaaag atgtgacaga
     7021 cgcgacggcg acaagcaaac atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc
     7081 aggtgatcgc tgatgtactg acaagcctcg cgtacccgat tatccatcgg tggatggagc
     7141 gactcgttaa tcgcttccat gtgccgcagt aacaattgct caagcagatt tatcgccagc
     7201 agctccgaat agcgcccttc cccttgcccg gcgttaatga tttgcccaaa caggtcgctg
     7261 aaatgcggct ggtgcgcttc atccgggcga aagaaccccg tattggcaaa tattgacggc
     7321 cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt aaacccactg gtgataccat
     7381 tcgcgagcct ccggatgacg accgtagtga tgaatctctc ctggcgggaa cagcaaaata
     7441 tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg accgcgaatg
     7501 gtgagattga gaatataacc tttcattccc agcggtcggt cgataaaaaa atcgagataa
     7561 ccgttggcct caatcggcgt taaacccgcc accagatggg cattaaacga gtatcccggc
     7621 agcaggggat cattttgcgc ttcagccata cttttcatac tcccgccatt cagagaagaa
     7681 accaattgtc catattgcat cagacattgc cgtcactgcg tcttttactg gctcttctcg
     7741 ctaaccaaac cggtaacccc gcttattaaa agcattctgt aacaaagcgg gaccaaagcc
     7801 atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca gaaaagtcca cattgattat
     7861 ttgcacggcg tcacactttg ctatgccata gcatttttat ccataagatt agcggatcct
     7921 acctgacgct ttttatcgca actctctact gtttctccat acccgttttt ttgggaattc
     7981 gagctctaag gaggttataa aaaatggata ttaatactga aactgagatc aagcaaaagc
     8041 attcactaac cccctttcct gttttcctaa tcagcccggc atttcgcggg cgatattttc
     8101 acagctattt caggagttca gccatgaacg cttattacat tcaggatcgt cttgaggctc
     8161 agagctgggc gcgtcactac cagcagctcg cccgtgaaga gaaagaggca gaactggcag
     8221 acgacatgga aaaaggcctg ccccagcacc tgtttgaatc gctatgcatc gatcatttgc
     8281 aacgccacgg ggccagcaaa aaatccatta cccgtgcgtt tgatgacgat gttgagtttc
     8341 aggagcgcat ggcagaacac atccggtaca tggttgaaac cattgctcac caccaggttg
     8401 atattgattc agaggtataa aacgaatgag tactgcactc gcaacgctgg ctgggaagct
     8461 ggctgaacgt gtcggcatgg attctgtcga cccacaggaa ctgatcacca ctcttcgcca
     8521 gacggcattt aaaggtgatg ccagcgatgc gcagttcatc gcattactga tcgttgccaa
     8581 ccagtacggc cttaatccgt ggacgaaaga aatttacgcc tttcctgata agcagaatgg
     8641 catcgttccg gtggtgggcg ttgatggctg gtcccgcatc atcaatgaaa accagcagtt
     8701 tgatggcatg gactttgagc aggacaatga atcctgtaca tgccggattt accgcaagga
     8761 ccgtaatcat ccgatctgcg ttaccgaatg gatggatgaa tgccgccgcg aaccattcaa
     8821 aactcgcgaa ggcagagaaa tcacggggcc gtggcagtcg catcccaaac ggatgttacg
     8881 tcataaagcc atgattcagt gtgcccgtct ggccttcgga tttgctggta tctatgacaa
     8941 ggatgaagcc gagcgcattg tcgaaaatac tgcatacact gcagaacgtc agccggaacg
     9001 cgacatcact ccggttaacg atgaaaccat gcaggagatt aacactctgc tgatcgccct
     9061 ggataaaaca tgggatgacg acttattgcc gctctgttcc cagatatttc gccgcgacat
     9121 tcgtgcatcg tcagaactga cacaggccga agcagtaaaa gctcttggat tcctgaaaca
     9181 gaaagccgca gagcagaagg tggcagcatg acaccggaca ttatcctgca gcgtaccggg
     9241 atcgatgtga gagctgtcga acagggggat gatgcgtggc acaaattacg gctcggcgtc
     9301 atcaccgctt cagaagttca caacgtgata gcaaaacccc gctccggaaa gaagtggcct
     9361 gacatgaaaa tgtcctactt ccacaccctg cttgctgagg tttgcaccgg tgtggctccg
     9421 gaagttaacg ctaaagcact ggcctgggga aaacagtacg agaacgacgc cagaaccctg
     9481 tttgaattca cttccggcgt gaatgttact gaatccccga tcatctatcg cgacgaaagt
     9541 atgcgtaccg cctgctctcc cgatggttta tgcagtgacg gcaacggcct tgaactgaaa
     9601 tgcccgttta cctcccggga tttcatgaag ttccggctcg gtggtttcga ggccataaag
     9661 tcagcttaca tggcccaggt gcagtacagc atgtgggtga cgcgaaaaaa tgcctggtac
     9721 tttgccaact atgacccgcg tatgaagcgt gaaggcctgc attatgtcgt gattgagcgg
     9781 gatgaaaagt acatggcgag ttttgacgag atcgtgccgg agttcatcga aaaaatggac
     9841 gaggcactgg ctgaaattgg ttttgtattt ggggagcaat ggcgatgacg catcctcacg
     9901 ataatatccg ggtaggcgca atcactttcg tctactccgt tacaaagcga ggctgggtat
     9961 ttcccggcct ttctgttatc cgaaatccac tgaaagcaca gcggctggct gaggagataa
    10021 ataataaacg aggggctgta tgcacaaagc atcttctgtt gagttaagaa cgagtatcga
    10081 gatggcacat agccttgctc aaattggaat caggtttgtg ccaataccag tagaaacaga
    10141 cgaagaatcc atgggtatgg aca
//
