LOCUS       sequence-346137-       10336 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis-MBP tagged Mb2Cas12a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10336)
  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 10336)
  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-huMb2Cas12a
FEATURES             Location/Qualifiers
     source          1..10336
                     /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             5273..5320
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5327..5407
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             5408..5425
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5478..5496)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5492..5539
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      5576..6031
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5663..5682)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5873..5894
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6124..6939)
                     /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(6124..6939)
                     /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     6847..6866
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6931..6950
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      7061..7649
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7550..7569
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7803..7820
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    7835..7974
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(7900..7919)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8060..8082
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(8076..8267)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(9076..10158)
                     /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     10120..10139
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(10159..10236)
                     /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 ccctgttcca ggacttcacc cacctgtacc ccctgagcaa
     1561 gaccgtcaga ttcgagctga agcccatcgg ccggaccctg gaacacatcc acgccaagaa
     1621 ctttctgagc caggacgaga caatggccga catgtaccag aaagtgaaag tgatcctgga
     1681 cgactaccac cgggacttta tcgccgacat gatgggcgaa gtgaagctga ccaagctggc
     1741 cgagttctac gacgtgtacc tgaagttccg gaagaacccc aaggacgacg gcctgcagaa
     1801 gcagctgaaa gacctgcagg ctgtgctgcg gaaagaaagc gtgaagccta tcggaagcgg
     1861 cggcaagtac aagaccggct acgacagact gttcggcgcc aagctgttca aggacggcaa
     1921 agagctgggc gacctggcca agttcgtgat cgcccaggaa ggcgagagca gccctaagct
     1981 ggcccacctg gcccatttcg agaagttcag cacctacttc accggcttcc acgacaaccg
     2041 caagaacatg tacagcgacg aggacaagca caccgccatt gcctaccggc tgatccacga
     2101 gaacctgccc cggttcatcg acaacctgca gatcctgacc accatcaagc agaagcacag
     2161 cgccctgtac gaccagatca tcaacgagct gaccgccagc ggcctggatg tgtctctggc
     2221 ctctcacctg gatggctacc acaagctgct gacacaggaa ggcatcaccg cctacaaccg
     2281 gatcatcgga gaagtgaacg gctacaccaa caagcacaac cagatctgcc acaagagcga
     2341 gcggattgcc aagctgcggc ccctgcacaa gcagattctg agcgacggaa tgggcgtgtc
     2401 cttcctgccc agcaagttcg ccgacgacag cgagatgtgc caggccgtga atgagttcta
     2461 ccggcactac accgacgtgt tcgccaaggt gcagagcctg ttcgacggct tcgacgacca
     2521 ccagaaagac ggcatctacg tggaacacaa gaacctgaac gagctgtcca agcaggcctt
     2581 cggcgacttt gccctgctgg gaagagtgct ggacggctac tatgtggacg tcgtgaaccc
     2641 cgagttcaac gagagatttg ccaaggccaa gaccgacaac gccaaagcca agctgacaaa
     2701 agagaaggac aagttcatca agggcgtgca ctccctggct tctctggaac aggccatcga
     2761 gcaccacaca gccagacacg acgacgagtc tgtgcaggcc ggaaagctgg gccagtactt
     2821 caagcacgga ctggccggcg tggacaaccc catccagaag atccacaaca accactctac
     2881 aatcaagggc ttcctggaaa gagagcggcc tgccggcgaa agagccctgc ccaagatcaa
     2941 gagcggcaag aaccctgaga tgacccagct gagacagctg aaagagctgc tggacaacgc
     3001 cctgaacgtg gcccacttcg ccaaactgct gaccacaaag accaccctgg acaaccagga
     3061 tggcaacttc tacggcgagt tcggcgtgct gtacgatgag ctggccaaga tccctaccct
     3121 gtacaacaaa gtgcgggact acctgagcca gaagcccttc agcaccgaga agtacaagct
     3181 gaatttcggc aaccccaccc tgctgaacgg ctgggacctg aacaaagaga aagataactt
     3241 cggcgtgatc ctgcagaaag atggctgcta ctacctggct ctgctggata aggcccacaa
     3301 gaaggtgttc gataacgccc ccaacaccgg caagaatgtg tatcagaaaa tggtgtataa
     3361 gctgctgcct ggccccaaca agatgctgcc caaggtgttc ttcgccaaga gcaacctgga
     3421 ctactacaac cccagcgccg aactgctgga caaatacgcc aagggcacac acaagaaagg
     3481 cgacaacttc aacctgaagg actgccacgc cctgatcgat ttcttcaagg ccggcattaa
     3541 caagcacccc gagtggcagc acttcggctt caagttcagc cccaccagca gctaccggga
     3601 cctgagcgac ttctacagag aggtggaacc ccagggctac caagtgaagt tcgtggacat
     3661 caacgccgac tacatcgacg agctggtgga acagggcaag ctgtacctgt ttcagatcta
     3721 caacaaggac tttagcccca aggcccacgg caagcccaac ctgcacaccc tgtactttaa
     3781 ggccctgttc agcgaggaca acctggccga ccccatctac aaactgaacg gcgaggccca
     3841 gatcttctac cggaaggcca gcctggacat gaacgagaca accatccaca gagccggcga
     3901 ggtgctggaa aacaagaatc ccgacaaccc taagaaacgg cagttcgtgt acgacatcat
     3961 caaggacaag cggtacaccc aggataagtt catgctgcac gtgcccatca ccatgaactt
     4021 cggagtgcag ggcatgacca tcaaagagtt caacaaaaaa gtgaaccaga gcatccagca
     4081 gtacgacgaa gtgaatgtga ttggcatcga ccggggcgag cggcatctgc tgtatctgac
     4141 cgtgatcaac agcaagggcg agattctgga acagagatcc ctgaacgaca tcaccaccgc
     4201 ctccgccaat ggcacccaag tgaccacccc ttaccacaag atcctggata agcgcgagat
     4261 cgagcggctg aacgccagag tgggatgggg agagatcgag acaatcaaag aactgaagtc
     4321 cggctacctg tcccacgtgg tgcatcagat caaccagctg atgctgaagt acaacgccat
     4381 cgtggtgctg gaagatctga attttggctt caagaggggc cggttcaagg tggaaaagca
     4441 gatctaccag aacttcgaga atgccctgat caagaaactg aaccacctgg tgctgaaaga
     4501 caaggccgac gacgagatcg gcagctacaa gaacgccctg cagctgacta acaacttcac
     4561 cgatctgaag tccattggca agcagaccgg ctttctgttc tacgtgcccg cctggaatac
     4621 cagcaagatc gaccccgaga caggcttcgt ggacctgctg aagcctagat acgagaatat
     4681 cgcccagtcc caggccttct tcggcaagtt cgacaagatc tgctacaaca ccgacaaggg
     4741 ctacttcgag ttccacatcg actacgccaa gttcaccgat aaggccaaaa acagccggca
     4801 gaagtgggct atctgcagcc acggcgacaa gagatacgtg tacgataaga ccgccaacca
     4861 gaacaagggc gctgccaagg gaatcaacgt gaacgacgaa ctgaaatccc tgttcgcccg
     4921 ctaccacatc aacgataagc agcccaatct cgtgatggac atctgccaga acaacgacaa
     4981 agagtttcac aagagcctga tgtgcctgct gaaaaccctg ctggccctgc ggtacagcaa
     5041 cgccagctcc gacgaggatt tcatcctgag ccccgtggcc aacgacgagg gcgtgttctt
     5101 caatagcgcc ctggccgatg acacccagcc ccagaacgct gatgccaacg gcgcctacca
     5161 cattgccctg aagggactgt ggctgctgaa tgagctgaag aactccgacg atctgaacaa
     5221 agtgaagctg gccatcgaca accagacctg gctgaatttc gcccagaaca gaaagaggcc
     5281 ggcggccacg aagaaggccg gccaggccaa gaagaagaag ggatcctacc catacgatgt
     5341 tccagattac gcttatccct acgacgtgcc tgattatgca tacccatatg atgtccccga
     5401 ctatgcgcac caccaccacc accactgaga tccggctgct aacaaagccc gaaaggaagc
     5461 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     5521 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg
     5581 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     5641 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     5701 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     5761 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     5821 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     5881 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     5941 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     6001 aattttaaca aactagtaac gtttacaatt tcaggtggca cttttcgggg aaatgtgcgc
     6061 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgaattaa
     6121 ttcttagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
     6181 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
     6241 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
     6301 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
     6361 gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg
     6421 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
     6481 ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat
     6541 cgaatgcaac cggcgcagga acactgccag cgcatcaaca atgttttcac ctgaatcagg
     6601 atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc
     6661 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
     6721 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
     6781 aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc
     6841 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
     6901 cggcctagag caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt
     6961 tatgtaagca gacagtttta ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc
     7021 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     7081 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     7141 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     7201 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     7261 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     7321 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     7381 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     7441 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     7501 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     7561 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     7621 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     7681 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     7741 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     7801 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     7861 atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc
     7921 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     7981 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     8041 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     8101 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
     8161 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
     8221 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
     8281 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
     8341 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
     8401 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     8461 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     8521 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
     8581 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
     8641 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
     8701 ctcaacgaca ggagcacgat catgcgcacc cgtggggccg ccatgccggc gataatggcc
     8761 tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc gagggcgtgc
     8821 aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc
     8881 tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat gataaagaag
     8941 acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga gctgactggg
     9001 ttgaaggctc tcaagggcat cggtcgagat cccggtgcct aatgagtgag ctaacttaca
     9061 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     9121 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt
     9181 tcttttcacc agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag
     9241 ttgcagcaag cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt
     9301 taacggcggg atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc
     9361 cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc
     9421 gttggcaacc agcatcgcag tgggaacgat accctcattc agcatttgca tggtttgttg
     9481 aaaaccggac atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg
     9541 agtgagatat ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc
     9601 cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt
     9661 accgtcttca tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa
     9721 taacgccgga acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg
     9781 atagttaatg atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca
     9841 ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc
     9901 gcgagattta atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc
     9961 aacgccaatc agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta
    10021 attcagctcc gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc
    10081 ctggttcacc acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta
    10141 taacgttact ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc
    10201 cataccgcga aaggttttgc gccattcgat ggtgtccggg atctcgacgc tctcccttat
    10261 gcgactcctg cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg
    10321 caaggaatgg tgcatg
//
