LOCUS       sequence-345928-       10276 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis-MBP tagged BfCas12a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10276)
  AUTHORS   Nguyen LT,  Macaluso NC,  Jain PK
  TITLE     A Combinatorial Approach towards Adaptability of 22 Functional 
            Cas12a Orthologs for Nucleic Acid Detection in Clinical Samples
  JOURNAL   medRxiv 2021.07.21
REFERENCE   2  (bases 1 to 10276)
  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: "medRxiv 
            2021.07.21"
COMMENT     Sequence Label: 6xHis-MBP-huBfCas12a
FEATURES             Location/Qualifiers
     source          1..10276
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     106..125
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     209..228
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        209..227
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    228..252
                     /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)."
     RBS             267..289
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             308..325
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             335..1435
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1409..1432
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1448..1468
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1478..1498
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5213..5260
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5267..5347
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             5348..5365
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5418..5436)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5432..5479
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      5516..5971
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5603..5622)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5813..5834
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6064..6879)
                     /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(6064..6879)
                     /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     6787..6806
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6871..6890
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      7001..7589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7490..7509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7743..7760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    7775..7914
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(7840..7859)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8000..8022
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(8016..8207)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(9016..10098)
                     /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     10060..10079
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(10099..10176)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa
       61 acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat
      121 ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta
      181 gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg
      241 gataacaatt cccctctaga aataattttg tttaacttta agaaggagat ataccatggg
      301 ttcttctcac catcaccatc accatggttc ttctatgaaa atcgaagaag gtaaactggt
      361 aatctggatt aacggcgata aaggctataa cggtctcgct gaagtcggta agaaattcga
      421 gaaagatacc ggaattaaag tcaccgttga gcatccggat aaactggaag agaaattccc
      481 acaggttgcg gcaactggcg atggccctga cattatcttc tgggcacacg accgctttgg
      541 tggctacgct caatctggcc tgttggctga aatcaccccg gacaaagcgt tccaggacaa
      601 gctgtatccg tttacctggg atgccgtacg ttacaacggc aagctgattg cttacccgat
      661 cgctgttgaa gcgttatcgc tgatttataa caaagatctg ctgccgaacc cgccaaaaac
      721 ctgggaagag atcccggcgc tggataaaga actgaaagcg aaaggtaaga gcgcgctgat
      781 gttcaacctg caagaaccgt acttcacctg gccgctgatt gctgctgacg ggggttatgc
      841 gttcaagtat gaaaacggca agtacgacat taaagacgtg ggcgtggata acgctggcgc
      901 gaaagcgggt ctgaccttcc tggttgacct gattaaaaac aaacacatga atgcagacac
      961 cgattactcc atcgcagaag ctgcctttaa taaaggcgaa acagcgatga ccatcaacgg
     1021 cccgtgggca tggtccaaca tcgacaccag caaagtgaat tatggtgtaa cggtactgcc
     1081 gaccttcaag ggtcaaccat ccaaaccgtt cgttggcgtg ctgagcgcag gtattaacgc
     1141 cgccagtccg aacaaagagc tggcaaaaga gttcctcgaa aactatctgc tgactgatga
     1201 aggtctggaa gcggttaata aagacaaacc gctgggtgcc gtagcgctga agtcttacga
     1261 ggaagagttg gcgaaagatc cacgtattgc cgccactatg gaaaacgccc agaaaggtga
     1321 aatcatgccg aacatcccgc agatgtccgc tttctggtat gccgtgcgta ctgcggtgat
     1381 caacgccgcc agcggtcgtc agactgtcga tgaagccctg aaagacgcgc agactaatgg
     1441 gatcgaggaa aacctgtact tccaatccaa tatggcccca aagaagaagc ggaaggtcgg
     1501 tatccacgga gtcccagcag cctactacga gagcctgacc aagctgtacc ccatcaagaa
     1561 aaccatccgg aacgagctgg tgcccatcgg caagaccctg gaaaacatca agaagaacaa
     1621 catcctggaa gccgacgagg accggaagat cgcctacatc agagtgaagg ccatcatgga
     1681 cgactaccac aagcggctga tcaacgaggc cctgagcggc ttcgccctga tcgacctgga
     1741 taaggccgcc aacctgtacc tgagcagaag caagagcgcc gacgacatcg agagcttcag
     1801 ccggttccag gacaagctgc ggaaggccat tgccaagaga ctgagagagc acgagaactt
     1861 cggcaagatc ggcaacaagg acatcatccc cctgctgcag aagctgagcg agaacgagga
     1921 cgattacaac gccctggaat ccttcaagaa cttctacacc tacttcgaga gctacaacga
     1981 cgtgcggctg aacctgtaca gcgacaaaga aaagtccagc accgtggcct accgcctgat
     2041 caatgagaac ctgcccagat tcctggacaa catccgggcc tacgacgccg tgcagaaggc
     2101 cggcatcaca agcgaggaac tgagcagcga ggcccaggac ggcctgttcc tcgtgaacac
     2161 cttcaacaac gtgctgatcc aggatggcat caacacctac aacgaggata tcggcaagct
     2221 gaacgtggcc atcaatctgt acaaccagaa aaacgccagc gtgcagggct tcagaaaggt
     2281 gcccaagatg aaggtgctgt acaagcagat cctgagcgac cgggaagagt ccttcatcga
     2341 cgagttcgag agcgacaccg agctgctgga cagcctggaa agccactacg ccaatctggc
     2401 caagtacttc ggcagcaaca aggtgcagct gctgttcacc gccctgcgcg agtctaaggg
     2461 cgtgaacgtg tacgtgaaga acgatatcgc caagaccagc ttcagcaacg tggtgttcgg
     2521 cagctggtcc cggatcgacg agctgattaa cggcgagtac gacgacaaca acaatcggaa
     2581 gaaggacgag aagtactacg acaagaggca gaaagagctg aagaaaaaca agagctacac
     2641 catcgagaag atcatcaccc tgagcaccga ggacgtggac gtgatcggca agtatatcga
     2701 gaagctggaa agcgacatcg acgatatccg gttcaagggc aagaatttct atgaggccgt
     2761 gctgtgcggc cacgaccgca gcaagaagct gtccaagaac aagggcgccg tggaagccat
     2821 caagggctac ctggacagcg tgaaggactt cgagcgggac ctgaagctga tcaatggcag
     2881 cggccaggaa ctggaaaaga acctggtggt gtacggcgag caggaagccg tgctgagcga
     2941 actgagcggc atcgacagcc tgtacaacat gacccggaac tacctgacaa agaagccctt
     3001 cagcaccgag aagattaagc tgaacttcaa caagcccacc ttcctggacg gctgggacta
     3061 cggcaacgag gaagcctacc tgggcttctt catgatcaaa gagggcaact actttctggc
     3121 cgtgatggac gccaactgga acaaagagtt cagaaacatc cccagcgtgg acaagagcga
     3181 ctgctacaag aaagtgatct ataagcagat cagcagcccc gagaagtcca tccagaatct
     3241 gatggtcatc gatggcaaga ccgtgaagaa gaacggccgg aaagagaaag aaggcatcca
     3301 cagcggcgag aacctgattc tggaagaact gaagaacacc tacctgccca agaagatcaa
     3361 cgacatccgg aagcggcgga gctacctgaa cggcgacacc ttctccaaga aggacctgac
     3421 cgagttcatc ggctactaca aacagcgcgt gatcgagtac tacaacggct acagcttcta
     3481 cttcaagtcc gacgacgact acgccagctt caaagaattt caggaagatg tgggccggca
     3541 ggcctaccag atcagctatg tggatgtgcc cgtgtccttc gtggacgacc tgatcaacag
     3601 cggcaaactg tacctgttcc gggtgtacaa caaggatttc agcgagtaca gcaagggcag
     3661 actgaacctg cacaccctgt acttcaagat gctgttcgac gagcgcaacc tgaagaatgt
     3721 ggtgtacaag ctgaatggcc aggccgaggt gttctaccgg cccagctcta tcaagaaaga
     3781 ggaactgatc gtgcacagag ccggcgagga aatcaagaac aaaaacccca agagagccgc
     3841 ccagaagccc accagaaggc tggactacga cattgtgaag gaccggcggt acagccagga
     3901 taagttcatg ctgcacacca gcatcatcat gaactttggc gccgaggaaa atgtgtcctt
     3961 taacgacatc gtgaacggcg tgctgcggaa tgaggacaaa gtgaatgtga tcggaatcga
     4021 ccggggcgag cggaacctgc tgtacgtggt cgtgattgac cccgagggaa agatcctgga
     4081 acagcggagc ctgaactgca tcaccgacag caacctggac atcgaaaccg attaccacag
     4141 actgctggac gagaaagaga gcgatagaaa gatcgccaga cgggactgga ccacaatcga
     4201 gaacatcaaa gaactgaagg ccggctacct gagccaggtg gtgcacattg tggccgaact
     4261 ggtgctgaag tacaacgcca tcatctgcct ggaagatctg aatttcggct tcaagcgggg
     4321 cagacagaaa gtggaaaaac aggtgtacca gaaatttgag aagatgctga ttgacaaact
     4381 gaattacctc gtgatggata agagccggga acagctgtcc cctgagaaga tctctggcgc
     4441 cctgaacgct ctgcagctga cccccgactt caagagcttc aaggtgctgg gaaagcagac
     4501 cggcatcatc tactacgtgc ccgcctacct gaccagcaag atcgacccca tgaccggctt
     4561 tgccaatctg ttctacgtga agtatgagaa tgtggacaag gccaaagagt ttttcagcaa
     4621 gttcgacagc atcaagtata acaaggacgg caagaactgg aataccaagg gctatttcga
     4681 gttcgccttc gactacaaga agttcaccga tagagcctac ggccgggtgt ccgagtggac
     4741 cgtgtgtaca gtgggcgaga gaatcatcaa gttcaaaaac aaagagaaga acaatagcta
     4801 cgatgacaaa gtgatcgatc tgaccaacag cctgaaagaa ctgttcgact cctacaaagt
     4861 gacctacgag tctgaggtgg acctgaaaga cgctatcctg gccatcgacg accccgcctt
     4921 ctaccgcgat ctgacaagac ggctgcagca gaccctgcag atgcggaaca gctcctgcga
     4981 cggcagccgg gactacatca tcagccctgt gaagaactcc aagggcgagt tcttctgctc
     5041 cgacaacaat gacgacacca cccccaacga cgccgacgcc aatggcgcct tcaatatcgc
     5101 cagaaagggc ctgtgggtgc tgaacgagat cagaaacagc gaggaaggct ccaagatcaa
     5161 cctggccatg agcaacgccc agtggctgga atacgcccag gacaacacca tcaagaggcc
     5221 ggcggccacg aagaaggccg gccaggccaa gaagaagaag ggatcctacc catacgatgt
     5281 tccagattac gcttatccct acgacgtgcc tgattatgca tacccatatg atgtccccga
     5341 ctatgcgcac caccaccacc accactgaga tccggctgct aacaaagccc gaaaggaagc
     5401 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     5461 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg
     5521 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     5581 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     5641 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     5701 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     5761 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     5821 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     5881 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     5941 aattttaaca aactagtaac gtttacaatt tcaggtggca cttttcgggg aaatgtgcgc
     6001 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgaattaa
     6061 ttcttagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
     6121 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
     6181 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
     6241 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
     6301 gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg
     6361 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
     6421 ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat
     6481 cgaatgcaac cggcgcagga acactgccag cgcatcaaca atgttttcac ctgaatcagg
     6541 atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc
     6601 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
     6661 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
     6721 aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc
     6781 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
     6841 cggcctagag caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt
     6901 tatgtaagca gacagtttta ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc
     6961 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     7021 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     7081 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     7141 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     7201 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     7261 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     7321 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     7381 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     7441 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     7501 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     7561 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     7621 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     7681 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     7741 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     7801 atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc
     7861 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     7921 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     7981 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     8041 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
     8101 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
     8161 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
     8221 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
     8281 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
     8341 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     8401 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     8461 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
     8521 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
     8581 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
     8641 ctcaacgaca ggagcacgat catgcgcacc cgtggggccg ccatgccggc gataatggcc
     8701 tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc
     8761 aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc
     8821 tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag
     8881 acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg
     8941 ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca
     9001 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     9061 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt
     9121 tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag
     9181 ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt
     9241 taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc
     9301 cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc
     9361 gttggcaacc agcatcgcag tgggaacgat accctcattc agcatttgca tggtttgttg
     9421 aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg
     9481 agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc
     9541 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
     9601 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
     9661 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
     9721 atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca
     9781 ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc
     9841 gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc
     9901 aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta
     9961 attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc
    10021 ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta
    10081 taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc
    10141 cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat
    10201 gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg
    10261 caaggaatgg tgcatg
//
