LOCUS       sequence-385919-        9670 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Engineered BrCas12b in Bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9670)
  AUTHORS   Nguyen LT, Rananaware SR, Yang LG, Macaluso NC, Ocana-Ortiz JE, 
            Meister KS, Pizzano BLM, Sandoval LSW, Hautamaki RC, Fang ZR, Joseph
            SM, Shoemaker GM, Carman DR, Chang L, Rakestraw NR, Zachary JF, 
            Guerra S, Perez A, Jain PK
  TITLE     Engineering highly thermostable Cas12b via de novo structural 
            analyses for one-pot detection of nucleic acids.
  JOURNAL   Cell Rep Med. 2023 May 16;4(5):101037. doi: 
            10.1016/j.xcrm.2023.101037. Epub 2023 May 8.
  PUBMED    37160120
REFERENCE   2  (bases 1 to 9670)
  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: "Cell Rep 
            Med."; date: "2023-05-16"; volume: "4(5):101037. doi"; pages: " 
            10.1016/j.xcrm.2023.101037. Epub 2023 May 8"
COMMENT     Sequence Label: 6xHis-MBP-BrCas12b-RFND
FEATURES             Location/Qualifiers
     source          1..9670
                     /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             4742..4759
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4812..4830)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4826..4873
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4910..5365
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4997..5016)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5207..5228
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5458..6273)
                     /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(5458..6273)
                     /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     6181..6200
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6265..6284
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6395..6983
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6884..6903
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7137..7154
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    7169..7308
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(7234..7253)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7394..7416
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(7410..7601)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(8410..9492)
                     /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     9454..9473
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(9493..9570)
                     /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 tatgcccgta aggtcattta aagttaagct
     1501 agtgactcgg tctggcgatg cggagcacat gctgcaactg cgtagggggc tgtggaaaac
     1561 ccatgaaatc gtaaatcagg gcattgcgta ctacatgaac aagttggcgc tgatgcgtca
     1621 agaaccgtat gctggtaaaa gccgtgaggt tgttcgtttg gagctgctgc atagcctgcg
     1681 tgcacagcag aaacgtaata actggaccgg cgacgccggt acggatgatg aaattctgaa
     1741 cctgagccgt cgcctgtacg agttgctcgt gccgagcgcg atcggcgaga agggcgatgc
     1801 gcagatgtta tctcgcaagt ttctgtcgcc gctggttgat ccgaattctg aaggtggtaa
     1861 aggcacggcg aagtccggcc gtaagccgcg ttggatgaaa atgcgggagg agggtcatcc
     1921 ggattgggaa gctgaacgcg agaaggacga agcaaagaaa gcagcggacc cgactgcgtc
     1981 tattctgaat gatttggaag cgtttggtct gcgtccgctg ttcccgctgt tcaccgacga
     2041 gcaaaaaggc atccagtggc tgccgaaaca aaaacgtcag ttcgtgcgta cctgggatcg
     2101 tgatatgttt cagcaggctc tggagcgtat gctgtcctgg gagtcctgga accgccgtgt
     2161 cgccgaggaa tatcagaagc tgcaggctca acgtgacgag ttgtacgcaa aatatctggc
     2221 cgatggtggt gcctggctgg aagcgttaca aagctttgag aagcagcgtg aagttgaact
     2281 ggctgaggag tcttttgccg ctaaatccga atatctgatc acccgacgac aaattcgcgg
     2341 ttggaaacag gtgtacgaaa agtggtcaca actgccagag cacgccgccc aagaacaatt
     2401 ctggcaagtt gtagcggacg tacagacgtc tctgcctggt gcgtttggtg acccaaaagt
     2461 ttaccagttt ctgagccagc cggagcatca ccacatctgg cgcggttacc cgaaccgact
     2521 attccactat tctgattaca acggtgttcg caagaagctg caaagagcgc gtcacgatgc
     2581 gacttttacc ttgccggacc cggttgaaca cccgctttgg atcagattcg atgcgcgtgg
     2641 tggtaatatt catgattatg agatcagcca gaatggcaag caatatcaag ttaccttctc
     2701 ccgcctcctg tggcctgaaa atgaaacctg ggtggagcgt gaaaacgtta ccgttgcaat
     2761 cggggcaagc cagcagctta agcgtcagat tcggttggac ggctacgccg ataaaaagca
     2821 gaaggtacgt tatcgtgact actctagcgg catcgagctg accggtgttc tgggtggtgc
     2881 gaagatccag ttcgaccgtc gtcacctgcg caaagcaagt aatcgtttag ccgacggtga
     2941 aaccggccct gtgtacctga acgtggtggt cgatatcgag ccgtttctgg ctatgcgcaa
     3001 cggtcgtcta caaacgccga ttggccaagt gctgcaagtc gtgaccaagg actggccgaa
     3061 ggtgacgggt tataaaccgg ctgagctgat cagctggatt caaaacagcc cgctggcggt
     3121 gggtacaggc gtcaacacca ttgaggcggg tatgcgcgtt atgagtgttg atctgggtca
     3181 aagaagcgca gctgctgtga gcattttcga ggtcatgcgc cagaaaccgg cggaacagga
     3241 aaccaaactc ttctatccga ttgctgtgac cggtctgtat gcggtgcatc gtcgtagttt
     3301 gctgctgcgt cttccaggcg agaaaattag cgacgaaatc gaacagcaac gtaaaatcag
     3361 agcccatgct cgtagcttgg tgcgctacca gatccgtctg cttgcggacg tactgcgtct
     3421 ccacacccgt ggcaccgcgg aacagagacg tgcgaaactg gatgaattgc tggctaccct
     3481 gcaaacgaaa caagagttag atcagaagtt atggcaaact gagttggaaa agctgttcga
     3541 ctacattcac gaaccggcgg agcgctggca gcaggcgctg gtggcggcgc accgtacctt
     3601 ggagccggtt atcggtcagg cagttcgtca ttggcgtaag tccctccgca ttgatcgtaa
     3661 gggcttggcg ggcatgagca tgtggaacat tgaagagttg gaggaaaccc ggaagttgtt
     3721 gatcgcatgg agcaaacatt ccagggtccc cggcgagccg aaccgtctgg acaaagaaga
     3781 aacgtttgca ccgcagcagc tccaacatat ccaaaacgtc aaagacgacc gcttgaagca
     3841 gatggcaaat ctgctcgtga tgaccgcctt gggctacaag tacgatgagg ccgagaaaca
     3901 atggaaagag gcgtatccgg cgtgtcagat gattctgttc gaagatctaa gccgctaccg
     3961 ctttgcgctg gaccgaccgc gtcgcgagaa caaccggtta atgaaatggg cacacagaag
     4021 tatcccgcgt ctggtttatc tgcaaggtga attgttcggc atccaggtgg gtgtggttta
     4081 ttcggcgtat actagccgtt tccatgctaa aactggtgcg ccaggcattc gctgccacgc
     4141 gctgaaagaa gaagacctgc aaccgaacag ctacgtggtg aagcaactga ttaaagacgg
     4201 ctttatccgt gaggaccaga ccggttccct taagccgggt cagattgtac cgtggtctgg
     4261 cggtgaactg ttcgttacct tagccgaccg ttccggcagc cgtttggccg tgatccacgc
     4321 cgacatcaat gcagcgcaga atctgcaaaa gagattctgg caacagaaca ccgagatctt
     4381 tcgtgtgccg tgcaaagtga ctacgtccgg tctgataccg gcatacgaca agatgaaaaa
     4441 gttattcggc aaaggctatt tcgcaaagat caaccagacg gacaccagcg aagtgtacgt
     4501 ttgggaacac agcgctaaga tgaagggcaa aacgaccccg gcggacccag cagaagaagg
     4561 agtttttgat gagtccctga ccgacgagat ggaagagctt gaagacagcc aagagggtta
     4621 caagaccctg ttccgtgatc cgtcgggctt tttttggtcg agcgatcgct ggctgccgca
     4681 gaaagagttc tggttttggg ttaaacgccg tatcgagaag aaactgcgcg agcagctcca
     4741 gcaccaccac caccaccact gagatccggc tgctaacaaa gcccgaaagg aagctgagtt
     4801 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     4861 gaggggtttt ttgctgaaag gaggaactat atccggattg gcgaatggga cgcgccctgt
     4921 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4981 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     5041 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     5101 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     5161 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     5221 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     5281 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     5341 aacaaactag taacgtttac aatttcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5401 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgaa ttaattctta
     5461 gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc
     5521 atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag
     5581 gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat
     5641 taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga
     5701 atccggtgag aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc
     5761 attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc
     5821 ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg
     5881 caaccggcgc aggaacactg ccagcgcatc aacaatgttt tcacctgaat caggatattc
     5941 ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc
     6001 aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag
     6061 tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa
     6121 ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt
     6181 atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta atcgcggcct
     6241 agagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta
     6301 agcagacagt tttattgttc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6361 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6421 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6481 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6541 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6601 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     6661 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     6721 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     6781 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     6841 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     6901 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     6961 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     7021 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     7081 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     7141 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     7201 gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     7261 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
     7321 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     7381 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     7441 gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc
     7501 tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata
     7561 aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg
     7621 ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg
     7681 gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta
     7741 tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca
     7801 gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg
     7861 gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt
     7921 catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt
     7981 atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac
     8041 gacaggagca cgatcatgcg cacccgtggg gccgccatgc cggcgataat ggcctgcttc
     8101 tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt
     8161 ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg
     8221 aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc
     8281 ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag
     8341 gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt
     8401 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     8461 atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt
     8521 caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag
     8581 caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg
     8641 cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc
     8701 aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc
     8761 aaccagcatc gcagtgggaa cgataccctc attcagcatt tgcatggttt gttgaaaacc
     8821 ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag
     8881 atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa
     8941 cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc
     9001 ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc
     9061 cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt
     9121 aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc
     9181 gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga
     9241 tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc
     9301 aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag
     9361 ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt
     9421 caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt
     9481 tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc
     9541 gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact
     9601 cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga
     9661 atggtgcatg
//
