LOCUS       sequence-346135-       10168 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis-MBP tagged PxCas12a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10168)
  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 10168)
  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-huPxCas12a
FEATURES             Location/Qualifiers
     source          1..10168
                     /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             5123..5143
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5150..5239
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             5240..5257
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5310..5328)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5324..5371
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      5408..5863
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5495..5514)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5705..5726
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5956..6771)
                     /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(5956..6771)
                     /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     6679..6698
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6763..6782
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6893..7481
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7382..7401
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7635..7652
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    7667..7806
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(7732..7751)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7892..7914
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(7908..8099)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(8908..9990)
                     /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     9952..9971
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(9991..10068)
                     /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 tatgatcatc ggccgcgact tcaacatgta
     1501 ctaccagaac ctgaccaaga tgtaccccat cagcaagacc ctgcgcaacg agctgatccc
     1561 cgtgggcaag accctggaga acatccgcaa gaacggcatc ctggaggccg acatccagcg
     1621 caaggccgac tacgagcacg tgaagaagct gatggacaac taccacaagc agctgatcaa
     1681 cgaggccctg cagggcgtgc acctgagcga cctgagcgac gcctacgacc tgtacttcaa
     1741 cctgagcaag gagaagaaca gcgtggacgc cttcagcaag tgccaggaca agctgcgcaa
     1801 ggagatcgtg agcttcctga agaaccacga gaacttcccc aagatcggca acaaggagat
     1861 catcaagctg atccagagcc tgaacgacaa cgacgccgac aacaacgccc tggacagctt
     1921 cagcaacttc tacacctact tcagcagcta caacgaggtg cgcaagaacc tgtacagcga
     1981 cgaggagaag agcagcaccg tggcctaccg cctgatcaac gagaacctgc ccaagagcct
     2041 ggacaacatc aaggcctacg ccatcgccaa gaaggccggc gtgcgcgccg agggcctgag
     2101 cgaggaggag caggactgcc tgttcatcat cgagaccttc gagcgcaccc tgacccagga
     2161 cggcatcgac aactacaacg ccgacatcgg caagctgaac accgccatca acctgtacaa
     2221 ccagcagaac aagaagcagg agggcttccg caaggtgccc cagatgaagt gcctgtacaa
     2281 gcagatcctg agcgaccgcg aggaggcctt catcgacgag ttcagcgacg acgaggacct
     2341 gatcaccaac atcgagagct tcgccgagaa catgaacgtg ttcctgaaca gcgagatcat
     2401 caccgacttc aagaacgccc tggtggagag cgacggcagc ctggtgtaca tcaagaacga
     2461 cgtgagcaag accctgttca gcaacatcgt gttcggcagc tggaacgcca tcgacgagaa
     2521 gctgagcgac gagtacgacc tggccaacag caagaagaag aaggacgaga agtactacga
     2581 gaagcgccag aaggagctga agaagaacaa gagctacgac ctggagacca tcatcggcct
     2641 gttcgacgac agcatcgacg tgatcggcaa gtacatcgag aagctggaga gcgacatcac
     2701 cgccatcgcc gaggccaaga acgacttcga cgagatcgtg ctgcgcaagc acgacaagaa
     2761 caagagcctg cgcaagaaca ccaacgccgt ggaggccatc aagagctacc tggacaccgt
     2821 gaaggacttc gagcgcgaca tcaagctgat caacggcagc ggccaggagg tggagaagaa
     2881 cctggtggtg tacgccgagc aggagaacat cctggccgag atcaagaacg tggacagcct
     2941 gtacaacatg agccgcaact acctgaccca gaagcccttc agcaccgaga agttcaagct
     3001 gaacttcgag aaccccaccc tgctgaacgg ctgggaccgc aacaaggaga aggactacct
     3061 gggcatcctg ttcgagaagg agggcatgta ctacctgggc atcatcaaca acaaccaccg
     3121 caagatcttc gagaacgaga agctgtgcac cggcaaggag agctgcttca acaagatcgt
     3181 gtacaagcag atcagcaacg ccgccaagta cctgagcagc aagcagatca acccccagaa
     3241 cccccccaag gagatcgccg agatcctgct gaagcgcaag gccgacagca gcagcctgag
     3301 ccgcaaggag accgagctgt tcatcgacta cctgaaggac gacttcctgg tgaactaccc
     3361 catgatcatc aacagcgacg gcgagaactt cttcaacttc cacttcaagc aggccaagga
     3421 ctacggcagc ctgcaggagt tcttcaagga ggtggagcac caggcctaca gcctgaagac
     3481 ccgccccatc gacgacagct acatctaccg catgatcgac gagggcaagc tgtacctgtt
     3541 ccagatccac aacaaggact tcagccccta cagcaagggc aacctgaacc tgcacaccat
     3601 ctacctgcag atgctgttcg accagcgcaa cctgaacaac gtggtgtaca agctgaacgg
     3661 cgaggccgag gtgttctacc gccccgccag catcaacgac gaggaggtga tcatccacaa
     3721 ggccggcgag gagatcaaga acaagaacag caagcgcgcc gtggacaagc ccaccagcaa
     3781 gttcggctac gacatcatca aggaccgccg ctacagcaag gacaagttca tgctgcacat
     3841 ccccgtgacc atgaacttcg gcgtggacga gacccgccgc ttcaacgacg tggtgaacga
     3901 cgccctgcgc aacgacgaga aggtgcgcgt gatcggcatc gaccgcggcg agcgcaacct
     3961 gctgtacgtg gtggtggtgg acaccgacgg caccatcctg gagcagatca gcctgaacag
     4021 catcatcaac aacgagtaca gcatcgagac cgactaccac aagctgctgg acgagaagga
     4081 gggcgaccgc gaccgcgccc gcaagaactg gaccaccatc gagaacatca aggagctgaa
     4141 ggagggctac ctgagccagg tggtgaacgt gatcgccaag ctggtgctga agtacaacgc
     4201 catcatctgc ctggaggacc tgaacttcgg cttcaagcgc ggccgccaga aggtggagaa
     4261 gcaggtgtac cagaagttcg agaagatgct gatcgacaag ctgaactacc tggtgatcga
     4321 caagagccgc aagcaggaga agcccgagga gttcggcggc gccctgaacg ccctgcagct
     4381 gaccagcaag ttcaccagct tcaaggacat gggcaagcag accggcatca tctactacgt
     4441 gcccgcctac ctgaccagca agatcgaccc caccaccggc ttcgccaacc tgttctacgt
     4501 gaagtacgag aacgtggaga aggccaagga gttcttcagc cgcttcgaca gcatcagcta
     4561 caacaacgag agcggctact tcgagttcgc cttcgactac aagaagttca ccgaccgcgc
     4621 ctgcggcgcc cgcagccagt ggaccgtgtg cacctacggc gagcgcatca tcaagtaccg
     4681 caacgccgac aagaacaaca gcttcgacga caagaccatc gtgctgagcg aggagttcaa
     4741 ggagctgttc agcatctacg gcatcagcta cgaggacggc gccgagctga agaacaagat
     4801 catgagcgtg gacgaggccg acttcttccg ctgcctgacc ggcctgctgc agaagaccct
     4861 gcagatgcgc aacagcagca acgacggcac ccgcgactac atcatcagcc ccatcatgaa
     4921 cgaccgcggc gagttcttca acagcgaggc ctgcgacgcc agcaagccca aggacgccga
     4981 cgccaacggc gccttcaaca tcgcccgcaa gggcctgtgg gtgctggagc agatccgcaa
     5041 cacccccagc ggcgacaagc tgaacctggc catgagcaac gccgagtggc tggagtacgc
     5101 ccagcgcaac cagatcgcta gcccaaagaa gaagaggaaa gtctctagat acccttatga
     5161 tgttccagat tatgccggat acccatacga tgtccctgac tatgcaggct cctaccctta
     5221 tgacgtccca gactacgcgc accaccacca ccaccactga gatccggctg ctaacaaagc
     5281 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     5341 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc
     5401 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     5461 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     5521 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     5581 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     5641 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     5701 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     5761 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     5821 aaatttaacg cgaattttaa caaactagta acgtttacaa tttcaggtgg cacttttcgg
     5881 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5941 ctcatgaatt aattcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
     6001 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     6061 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     6121 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     6181 accatgagtg acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac
     6241 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     6301 attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt
     6361 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatgttttc
     6421 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
     6481 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     6541 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     6601 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
     6661 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     6721 ggaatttaat cgcggcctag agcaagacgt ttcccgttga atatggctca taacacccct
     6781 tgtattactg tttatgtaag cagacagttt tattgttcat gaccaaaatc ccttaacgtg
     6841 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6901 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6961 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     7021 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     7081 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     7141 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     7201 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     7261 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     7321 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     7381 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     7441 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     7501 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     7561 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     7621 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     7681 ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg
     7741 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
     7801 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     7861 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     7921 ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc
     7981 gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt
     8041 aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact
     8101 gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag
     8161 aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag
     8221 ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc
     8281 cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg
     8341 cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca
     8401 cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg
     8461 cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag
     8521 cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggggc cgccatgccg
     8581 gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga
     8641 gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag
     8701 cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc
     8761 atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag
     8821 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg
     8881 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     8941 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     9001 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     9061 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     9121 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
     9181 taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     9241 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg ataccctcat tcagcatttg
     9301 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     9361 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     9421 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     9481 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     9541 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     9601 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     9661 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     9721 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     9781 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     9841 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     9901 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     9961 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
    10021 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac
    10081 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
    10141 gcaccgccgc cgcaaggaat ggtgcatg
//
