LOCUS       sequence-346166-       10366 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis-MBP tagged Mb3Cas12a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10366)
  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 10366)
  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-huMb3Cas12a
FEATURES             Location/Qualifiers
     source          1..10366
                     /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             5303..5350
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5357..5437
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             5438..5455
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5508..5526)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5522..5569
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      5606..6061
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5693..5712)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5903..5924
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6154..6969)
                     /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(6154..6969)
                     /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     6877..6896
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6961..6980
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      7091..7679
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7580..7599
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7833..7850
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    7865..8004
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(7930..7949)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8090..8112
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(8106..8297)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(9106..10188)
                     /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     10150..10169
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(10189..10266)
                     /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 caagaccctg gaacacatcc acgccaagaa
     1621 ctttctgaac caggacgaga caatggccga catgtaccag aaagtgaagg ccatcctgga
     1681 cgactaccac cgggacttta tcgccgacat gatgggcgaa gtgaagctga ccaagctggc
     1741 cgagttctac gacgtgtacc tgaagttccg gaagaacccc aaggacgacg gcctgcagaa
     1801 gcagctgaaa gacctgcagg ctgtgctgcg gaaagaaatc gtgaagccta tcggaaacgg
     1861 cggcaagtac aaggccggct 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 gatcctggcc accatcaagc agaagcacag
     2161 cgccctgtac gaccagatca tcaacgagct gaccgccagc ggcctggatg tgtctctggc
     2221 ctctcacctg gatggctacc acaagctgct gacacaggaa ggcatcaccg cctacaacac
     2281 cctgctgggc ggaatctctg gcgaggccgg cagcagaaag atccagggca tcaatgaact
     2341 gatcaacagc caccacaacc agcactgcca caagagcgag cggattgcca agctgcggcc
     2401 cctgcacaag cagattctga gcgacggaat gggcgtgtcc ttcctgccca gcaagttcgc
     2461 cgacgacagc gaagtgtgcc aggccgtgaa tgagttctac cggcactacg ccgacgtgtt
     2521 cgccaaggtg cagagcctgt tcgacggctt cgacgactat cagaaagacg gcatctacgt
     2581 ggagtacaag aacctgaacg agctgtccaa gcaggccttc ggcgacttcg ctctgctggg
     2641 aagagtgctg gacggctact atgtggacgt cgtgaacccc gagttcaacg agagatttgc
     2701 caaggccaag accgacaacg ccaaagccaa gctgacaaaa gagaaggaca agttcatcaa
     2761 gggcgtgcac tccctggctt ctctggaaca ggccatcgag cactacaccg ccagacacga
     2821 cgacgagtct gtgcaggccg gaaagctggg ccagtacttc aagcacggac tggccggcgt
     2881 ggacaacccc atccagaaga tccacaacaa ccactctaca atcaagggct tcctggaaag
     2941 agagcggcct gccggcgaaa gagccctgcc caagatcaag agcgacaaga gccccgagat
     3001 cagacagctg aaagagctgc tggacaacgc cctgaacgtg gcccacttcg ccaaactgct
     3061 gaccaccaag accacactgc acaatcagga tggcaacttc tacggcgagt tcggagccct
     3121 gtatgatgag ctggccaaga tcgccaccct gtacaacaaa gtgcgggact acctgagcca
     3181 gaagcccttc agcaccgaga agtacaagct gaatttcggc aaccctaccc tgctgaacgg
     3241 ctgggacctg aacaaagaga aagataactt cggcgtgatc ctgcagaagg acggctgcta
     3301 ctacctggcc ctgctggata aggcccacaa gaaggtgttc gataacgccc ccaacaccgg
     3361 caagtctgtg tatcagaaga tgatctataa gctgctgcct ggccccaaca agatgctgcc
     3421 caaggtgttc ttcgccaaga gcaacctgga ctactacaac cccagcgccg aactgctgga
     3481 caaatacgcc cagggcacac acaagaaagg cgacaacttc aacctgaagg actgccacgc
     3541 cctgatcgat ttcttcaagg ccggaatcaa caagcacccc gagtggcagc acttcggctt
     3601 caagttcagc cccaccagca gctaccagga cctgagcgac ttctacagag aggtggaacc
     3661 ccagggctac caagtgaagt tcgtggacat caacgccgac tacatcaatg agctggtgga
     3721 acagggccag ctgtacctgt ttcagatcta caacaaggac tttagcccca aggcccacgg
     3781 caagcccaac ctgcacaccc tgtatttcaa ggccctgttt agcgaggaca acctcgtgaa
     3841 tcccatctac aaactgaacg gggaggccga gatcttctac cggaaggcct ccctggacat
     3901 gaacgagaca accatccaca gagccggcga ggtgctggaa aacaagaacc ctgacaaccc
     3961 taagaaacgg cagttcgtgt acgacatcat caaggacaag cggtacaccc aggataagtt
     4021 catgctgcac gtgcccatca ccatgaactt cggagtgcag ggcatgacca tcaaagagtt
     4081 caacaaaaaa gtgaaccaga gcatccagca gtacgacgaa gtgaacgtga tcggcatcga
     4141 ccggggcgag cggcatctgc tgtatctgac cgtgatcaac tccaagggcg agattctgga
     4201 acagagatcc ctgaacgaca tcaccaccgc ctccgccaac ggcacccaga tgaccacccc
     4261 ttaccacaag atcctggaca agcgcgagat cgagcggctg aacgccagag tgggatgggg
     4321 agagatcgag acaatcaaag aactgaagtc cggctacctg tcccacgtgg tgcatcagat
     4381 ctcccagctg atgctgaagt acaacgccat cgtggtgctg gaagatctga attttggctt
     4441 caagaggggc cggttcaagg tggaaaagca gatctaccag aacttcgaga atgccctgat
     4501 caagaaactg aaccacctgg tgctgaaaga caaggccgac gacgagatcg gcagctacaa
     4561 gaacgccctg cagctgacta acaacttcac cgatctgaag agtatcggca agcagaccgg
     4621 ctttctgttc tacgtgcccg cctggaatac cagcaagatc gaccccgaga caggcttcgt
     4681 ggacctgctg aagcctagat acgagaatat cgcccagagc caggccttct tcggcaagtt
     4741 cgacaagatc tgctacaacg ccgatagggg ctacttcgag ttccacatcg actacgccaa
     4801 gttcaatgac aaggccaaaa acagccggca gatctggaaa atctgcagcc acggcgataa
     4861 gcgctacgtg tacgataaga ccgccaacca gaacaagggc gccaccatcg gagtgaatgt
     4921 gaacgatgag ctgaagtccc tgttcacccg ctaccacatc aacgataagc agcccaatct
     4981 cgtgatggac atctgccaga acaacgacaa agagtttcac aagagcctga tgtatctgct
     5041 gaaaacactg ctggctctgc ggtacagcaa cgccagctcc gacgaggatt tcatcctgag
     5101 ccccgtggcc aacgacgagg gcgtgttctt caatagcgcc ctggccgacg atacccagcc
     5161 ccagaatgcc gatgccaacg gcgcctacca cattgccctg aagggactgt ggctgctgaa
     5221 cgaactgaag aacagcgacg atctgaacaa agtgaagctg gccatcgaca accagacctg
     5281 gctgaatttc gcccagaaca gaaagaggcc ggcggccacg aagaaggccg gccaggccaa
     5341 gaagaagaag ggatcctacc catacgatgt tccagattac gcttatccct acgacgtgcc
     5401 tgattatgca tacccatatg atgtccccga ctatgcgcac caccaccacc accactgaga
     5461 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
     5521 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
     5581 aactatatcc ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     5641 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     5701 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     5761 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     5821 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     5881 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     5941 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     6001 aatgagctga tttaacaaaa atttaacgcg aattttaaca aactagtaac gtttacaatt
     6061 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     6121 cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga gcatcaaatg
     6181 aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
     6241 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
     6301 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
     6361 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt
     6421 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
     6481 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc
     6541 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
     6601 cgcatcaaca atgttttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     6661 cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     6721 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     6781 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     6841 caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     6901 taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt cccgttgaat
     6961 atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga
     7021 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     7081 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     7141 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     7201 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     7261 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     7321 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     7381 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     7441 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     7501 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     7561 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     7621 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     7681 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     7741 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     7801 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     7861 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg
     7921 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     7981 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     8041 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     8101 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca
     8161 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg
     8221 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt
     8281 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg
     8341 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
     8401 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
     8461 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc
     8521 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt
     8581 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac
     8641 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca
     8701 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc
     8761 cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga
     8821 ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg
     8881 atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc
     8941 acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg
     9001 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
     9061 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
     9121 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     9181 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     9241 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     9301 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     9361 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
     9421 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     9481 accctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     9541 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     9601 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     9661 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     9721 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     9781 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     9841 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     9901 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     9961 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
    10021 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
    10081 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
    10141 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
    10201 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
    10261 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta
    10321 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatg
//
