LOCUS       sequence-346186-       10318 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis-MBP tagged PcCas12a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10318)
  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 10318)
  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-huPcCas12a
FEATURES             Location/Qualifiers
     source          1..10318
                     /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             5252..5299
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5306..5386
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             5390..5407
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5460..5478)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5474..5521
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      5558..6013
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5645..5664)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5855..5876
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6106..6921)
                     /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(6106..6921)
                     /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     6829..6848
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6913..6932
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      7043..7631
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7532..7551
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7785..7802
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    7817..7956
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(7882..7901)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8042..8064
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(8058..8249)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(9058..10140)
                     /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     10102..10121
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(10141..10218)
                     /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 tatggacagc ctgaaggatt tcaccaacct
     1501 gtaccccgtg tccaagacac tgcggtttga gctgaagcct gtgggcaaga ccctggagaa
     1561 tatcgagaag gccggcatcc tgaaggagga tgagcacaga gccgagagct accggagagt
     1621 gaagaagatc atcgatacat atcacaaggt gttcatcgac agctccctgg agaacatggc
     1681 caagatgggc atcgagaatg agatcaaggc catgctgcag tccttttgcg agctgtataa
     1741 gaaggaccac aggaccgagg gagaggacaa ggccctggat aagatcaggg ccgtgctgag
     1801 gggcctgatc gtgggagcct tcaccggcgt gtgcggccgg cgggagaaca cagtgcagaa
     1861 tgagaagtat gagagcctgt ttaaggagaa gctgatcaag gagatcctgc cagatttcgt
     1921 gctgtctaca gaggccgagt ccctgccctt ttctgtggag gaggccacca gaagcctgaa
     1981 ggagttcgac tcctttacat cttacttcgc cggcttttat gagaaccgga agaatatcta
     2041 ctctaccaag ccccagagca cagccatcgc ctatagactg atccacgaga acctgcctaa
     2101 gttcatcgat aatatcctgg tgtttcagaa gatcaaggag ccaatcgcca aggagctgga
     2161 gcacatcagg gcagacttca gcgccggcgg ctacatcaag aaggatgagc gcctggagga
     2221 catcttttcc ctgaactact atatccacgt gctgtctcag gccggcatcg agaagtacaa
     2281 tgccctgatc ggcaagatcg tgaccgaggg cgatggcgag atgaagggcc tgaacgagca
     2341 catcaacctg tataatcagc agaggggccg cgaggaccgg ctgccactgt tcagacccct
     2401 gtataagcag atcctgtctg atagggagca gctgtcctat ctgccagagt ctttcgagaa
     2461 ggacgaggag ctgctgaggg ccctgaagga gttttacgat cacatcgcag aggacatcct
     2521 gggaaggacc cagcagctga tgacaagcat ctccgagtac gatctgtccc ggatctatgt
     2581 gagaaacgat agccagctga ccgacatctc caagaagatg ctgggcgatt ggaatgccat
     2641 ctacatggcc cgggagagag cctatgacca cgagcaggcc cccaagcgca tcacagccaa
     2701 gtacgagagg gaccgcatca aggccctgaa gggcgaggag tctatcagcc tggccaacct
     2761 gaacagctgc atcgccttcc tggacaacgt gagggattgt cgcgtggaca cctatctgtc
     2821 tacactggga cagaaggagg gacctcacgg cctgagcaac ctggtggaga acgtgttcgc
     2881 ctcctaccac gaggccgagc agctgctgtc ttttccctat cctgaggaga acaatctgat
     2941 ccaggacaag gataacgtgg tgctgatcaa gaacctgctg gataatatca gcgacctgca
     3001 gaggttcctg aagccactgt ggggcatggg cgatgagccc gacaaggatg agaggtttta
     3061 cggcgagtac aattatatca ggggcgccct ggaccaggtc atccctctgt ataacaaggt
     3121 gcggaattat ctgacccgca agccatactc cacacgcaag gtgaagctga acttcggcaa
     3181 tagccagctg ctgtccggct gggataggaa caaggagaag gacaattctt gcgtgatcct
     3241 gcgcaagggc cagaacttct acctggccat catgaacaat cggcacaagc ggagcttcga
     3301 gaataagatg ctgcccgagt ataaggaggg cgagccttac ttcgagaaga tggattataa
     3361 gtttctgcca gaccccaaca agatgctgcc caaggtgttc ctgtctaaga agggcatcga
     3421 gatctacaag cctagcccaa agctgctgga gcagtatggc cacggcaccc acaagaaggg
     3481 cgataccttc agcatggacg atctgcacga gctgatcgac ttctttaagc actccatcga
     3541 ggcccacgag gattggaagc agttcggctt taagttcagc gacaccgcca catacgagaa
     3601 cgtgagcagc ttctaccggg aggtggagga ccagggctac aagctgtctt ttagaaaggt
     3661 gtccgagtct tacgtgtata gcctgatcga tcagggcaag ctgtacctgt tccagatcta
     3721 taacaaggac tttagccctt gttccaaggg caccccaaat ctgcacacac tgtactggcg
     3781 gatgctgttc gatgagagaa acctggccga cgtgatctat aagctggatg gcaaggccga
     3841 gatcttcttt cgggagaagt ccctgaagaa tgaccaccca acccaccctg caggcaagcc
     3901 catcaagaag aagagccggc agaagaaggg cgaggagagc ctgttcgagt acgatctggt
     3961 gaaggaccgg agatatacca tggataagtt tcagttccac gtgccaatca caatgaactt
     4021 taagtgctct gccggcagca aggtgaacga catggtgaat gcccacatca gggaggccaa
     4081 ggacatgcac gtgatcggca tcgatagggg cgagcgcaat ctgctgtata tctgcgtgat
     4141 cgacagccgc ggcaccatcc tggatcagat ctccctgaac acaatcaatg acatcgatta
     4201 tcacgatctg ctggagtcca gggacaagga tcgccagcag gagcacagga actggcagac
     4261 catcgagggc atcaaggagc tgaagcaggg ctacctgtct caggccgtgc accgcatcgc
     4321 cgagctgatg gtggcctata aggccgtggt ggccctggag gacctgaaca tgggcttcaa
     4381 gcggggcaga cagaaggtgg agagcagcgt gtaccagcag tttgagaagc agctgatcga
     4441 caagctgaat tatctggtgg ataagaagaa gcggcccgag gacatcggag gcctgctgag
     4501 agcctaccag ttcaccgccc ctttcaagag ctttaaggag atgggcaagc agaacggctt
     4561 tctgttctat atccctgcct ggaacacatc caatatcgac ccaaccacag gcttcgtgaa
     4621 cctgtttcac gtgcagtacg agaatgtgga taaggccaag agcttctttc agaagttcga
     4681 cagcatctcc tacaacccta agaaggattg gtttgagttc gcctttgact ataagaactt
     4741 caccaagaag gccgagggct ctaggagcat gtggattctg tgcacccacg gctcccggat
     4801 caagaacttc agaaattctc agaagaatgg ccagtgggat agcgaggagt ttgccctgac
     4861 cgaggccttc aagtccctgt ttgtgcggta cgagatcgat tataccgccg acctgaaaac
     4921 cgccatcgtg gacgagaagc agaaggattt ctttgtggac ctgctgaagc tgttcaagct
     4981 gaccgtgcag atgagaaact cctggaagga gaaggacctg gattacctga tctctccagt
     5041 ggccggcgcc gatggcaggt tctttgacac acgcgagggc aataagagcc tgcccaagga
     5101 cgcagatgca aacggagcct ataatatcgc cctgaagggc ctgtgggcac tgaggcagat
     5161 cagacagacc tccgagggcg gcaagctgaa gctggccatc tctaacaagg agtggctgca
     5221 gtttgtgcag gagagatcct acgagaagga caaaaggccg gcggccacga aaaaggccgg
     5281 ccaggcaaaa aagaaaaagg gatcctaccc atacgatgtt ccagattacg cttatcccta
     5341 cgacgtgcct gattatgcat acccatatga tgtccccgac tatgcctagc accaccacca
     5401 ccaccactga gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac
     5461 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     5521 gctgaaagga ggaactatat ccggattggc gaatgggacg cgccctgtag cggcgcatta
     5581 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     5641 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     5701 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     5761 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     5821 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     5881 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     5941 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaactagta
     6001 acgtttacaa tttcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     6061 tttttctaaa tacattcaaa tatgtatccg ctcatgaatt aattcttaga aaaactcatc
     6121 gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa
     6181 aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc
     6241 ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc
     6301 gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa
     6361 tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc
     6421 atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg
     6481 aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag
     6541 gaacactgcc agcgcatcaa caatgttttc acctgaatca ggatattctt ctaatacctg
     6601 gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat
     6661 aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc
     6721 atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc
     6781 gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca
     6841 tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctag agcaagacgt
     6901 ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag cagacagttt
     6961 tattgttcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     7021 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     7081 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     7141 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     7201 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     7261 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     7321 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     7381 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     7441 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     7501 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     7561 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     7621 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     7681 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     7741 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     7801 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     7861 accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac
     7921 actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct
     7981 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     8041 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg
     8101 cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg
     8161 tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg
     8221 ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc
     8281 atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat
     8341 gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg
     8401 gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt
     8461 ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct
     8521 gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct
     8581 caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca
     8641 ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg
     8701 atcatgcgca cccgtggggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt
     8761 ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca
     8821 agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag
     8881 agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg
     8941 acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
     9001 atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca
     9061 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     9121 gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac
     9181 gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac
     9241 gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca
     9301 tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc
     9361 ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc
     9421 agtgggaacg ataccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact
     9481 ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca
     9541 gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg
     9601 ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa
     9661 aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt
     9721 gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc
     9781 actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg
     9841 ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc
     9901 gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga
     9961 ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc
    10021 cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga
    10081 aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac
    10141 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
    10201 gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga
    10261 agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatg
//
