LOCUS       sequence-346188-       10504 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis-MBP tagged PdCas12a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10504)
  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 10504)
  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-huPdCas12a
FEATURES             Location/Qualifiers
     source          1..10504
                     /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             5441..5488
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5495..5575
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             5576..5593
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5646..5664)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5660..5707
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      5744..6199
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5831..5850)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6041..6062
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6292..7107)
                     /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(6292..7107)
                     /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     7015..7034
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7099..7118
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      7229..7817
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7718..7737
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7971..7988
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8003..8142
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8068..8087)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8228..8250
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(8244..8435)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(9244..10326)
                     /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     10288..10307
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(10327..10404)
                     /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 tatggagaac tatcaggagt tcaccaacct
     1501 gtttcagctg aataagacac tgagattcga gctgaagccc atcggcaaga cctgcgagct
     1561 gctggaggag ggcaagatct tcgccagcgg ctcctttctg gagaaggaca aggtgagggc
     1621 cgataacgtg agctacgtga agaaggagat cgacaagaag cacaagatct ttatcgagga
     1681 gacactgagc tccttctcta tcagcaacga tctgctgaag cagtactttg actgctataa
     1741 tgagctgaag gccttcaaga aggactgtaa gagcgatgag gaggaggtga agaaaaccgc
     1801 cctgcgcaac aagtgtacct ccatccagag ggccatgcgc gaggccatct ctcaggcctt
     1861 tctgaagagc ccccagaaga agctgctggc catcaagaac ctgatcgaga acgtgttcaa
     1921 ggccgacgag aatgtgcagc acttctccga gtttaccagc tatttctccg gctttgagac
     1981 aaacagagag aatttctact ctgacgagga gaagtccaca tctatcgcct ataggctggt
     2041 gcacgataac ctgcctatct tcatcaagaa catctacatc ttcgagaagc tgaaggagca
     2101 gttcgacgcc aagaccctga gcgagatctt cgagaactac aagctgtatg tggccggctc
     2161 tagcctggat gaggtgttct ccctggagta ctttaacaat accctgacac agaagggcat
     2221 cgacaactat aatgccgtga tcggcaagat cgtgaaggag gataagcagg agatccaggg
     2281 cctgaacgag cacatcaacc tgtataatca gaagcacaag gaccggagac tgcccttctt
     2341 tatctccctg aagaagcaga tcctgtccga tcgggaggcc ctgtcttggc tgcctgacat
     2401 gttcaagaat gattctgaag tgatcaaggc cctgaagggc ttctacatcg aggacggctt
     2461 tgagaacaat gtgctgacac ctctggccac cctgctgtcc tctctggata agtacaacct
     2521 gaatggcatc tttatccgca acaatgaggc cctgagctcc ctgtcccaga acgtgtatcg
     2581 gaatttttct atcgacgagg ccatcgatgc caacgccgag ctgcagacct tcaacaatta
     2641 cgagctgatc gccaatgccc tgcgcgccaa gatcaagaag gagacaaagc agggccggaa
     2701 gtctttcgag aagtacgagg agtatatcga taagaaggtg aaggccatcg acagcctgtc
     2761 catccaggag atcaacgagc tggtggagaa ttacgtgagc gagtttaact ctaatagcgg
     2821 caacatgcca agaaaggtgg aggactactt cagcctgatg aggaagggcg acttcggctc
     2881 caacgatctg atcgaaaata tcaagaccaa gctgagcgcc gcagagaagc tgctgggcac
     2941 aaagtaccag gagacagcca aggacatctt caagaaggat gagaactcca agctgatcaa
     3001 ggagctgctg gacgccacca agcagttcca gcactttatc aagccactgc tgggcacagg
     3061 cgaggaggca gatcgggacc tggtgttcta cggcgatttt ctgcccctgt atgagaagtt
     3121 tgaggagctg accctgctgt ataacaaggt gcggaataga ctgacacaga agccctattc
     3181 caaggacaag atccgcctgt gcttcaacaa gcctaagctg atgacaggct gggtggattc
     3241 caagaccgag aagtctgaca acggcacaca gtacggcggc tatctgtttc ggaagaagaa
     3301 tgagatcggc gagtacgatt attttctggg catctctagc aaggcccagc tgttcagaaa
     3361 gaacgaggcc gtgatcggcg actacgagag gctggattac tatcagccaa aggccaatac
     3421 catctacggc tctgcctatg agggcgagaa cagctacaag gaggacaaga agcggctgaa
     3481 caaagtgatc atcgcctata tcgagcagat caagcagaca aacatcaaga agtctatcat
     3541 cgagtccatc tctaagtatc ctaatatcag cgacgatgac aaggtgaccc catcctctct
     3601 gctggagaag atcaagaagg tgtctatcga cagctacaac ggcatcctgt ccttcaagtc
     3661 ttttcagagc gtgaacaagg aagtgatcga taacctgctg aaaaccatca gccccctgaa
     3721 gaacaaggcc gagtttctgg acctgatcaa taaggattat cagatcttca ccgaggtgca
     3781 ggccgtgatc gacgagatct gcaagcagaa aaccttcatc tactttccaa tctccaacgt
     3841 ggagctggag aaggagatgg gcgataagga caagcccctg tgcctgttcc agatcagcaa
     3901 taaggatctg tccttcgcca agacctttag cgccaacctg cggaagaaga gaggcgccga
     3961 gaatctgcac acaatgctgt ttaaggccct gatggagggc aaccaggata atctggacct
     4021 gggctctggc gccatcttct acagagccaa gagcctggac ggcaacaagc ccacacaccc
     4081 tgccaatgag gccatcaagt gtaggaacgt ggccaataag gataaggtgt ccctgttcac
     4141 ctacgacatc tataagaaca ggcgctacat ggagaataag ttcctgtttc acctgagcat
     4201 cgtgcagaac tataaggccg ccaatgactc cgcccagctg aacagctccg ccaccgagta
     4261 tatcagaaag gccgatgacc tgcacatcat cggcatcgat aggggcgagc gcaatctgct
     4321 gtactattcc gtgatcgata tgaagggcaa catcgtggag caggactctc tgaatatcat
     4381 caggaacaat gacctggaga cagattacca cgacctgctg gataagaggg agaaggagcg
     4441 caaggccaac cggcagaatt gggaggccgt ggagggcatc aaggacctga agaagggcta
     4501 cctgagccag gccgtgcacc agatcgccca gctgatgctg aagtataacg ccatcatcgc
     4561 cctggaggat ctgggccaga tgtttgtgac ccgcggccag aagatcgaga aggccgtgta
     4621 ccagcagttc gagaagagcc tggtggataa gctgtcctac ctggtggaca agaagcggcc
     4681 ttataatgag ctgggcggca tcctgaaggc ctaccagctg gcctctagca tcaccaagaa
     4741 caattctgac aagcagaacg gcttcctgtt ttatgtgcca gcctggaata caagcaagat
     4801 cgatcccgtg accggcttta cagacctgct gcggcccaag gccatgacca tcaaggaggc
     4861 ccaggacttc tttggcgcct tcgataacat ctcttacaat gacaagggct atttcgagtt
     4921 tgagacaaac tacgacaagt ttaagatcag aatgaagagc gcccagacca ggtggacaat
     4981 ctgcaccttc ggcaatcgga tcaagagaaa gaaggataag aactactgga attatgagga
     5041 ggtggagctg accgaggagt tcaagaagct gtttaaggac agcaacatcg attacgagaa
     5101 ctgtaatctg aaggaggaga tccagaacaa ggacaatcgc aagttctttg atgacctgat
     5161 caagctgctg cagctgacac tgcagatgcg gaactccgat gacaagggca atgattatat
     5221 catctctcct gtggccaacg ccgagggcca gttctttgac tcccgcaatg gcgataagaa
     5281 gctgccactg gatgcagacg caaacggagc ctacaatatc gcccgcaagg gcctgtggaa
     5341 catccggcag atcaagcaga ccaagaacga caagaagctg aatctgagca tctcctctac
     5401 agagtggctg gatttcgtgc gggagaagcc ttacctgaag aaaaggccgg cggccacgaa
     5461 aaaggccggc caggcaaaaa agaaaaaggg atcctaccca tacgatgttc cagattacgc
     5521 ttatccctac gacgtgcctg attatgcata cccatatgat gtccccgact atgcgcacca
     5581 ccaccaccac cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     5641 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     5701 ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc
     5761 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     5821 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     5881 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     5941 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     6001 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     6061 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     6121 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     6181 ctagtaacgt ttacaatttc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     6241 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgaattaatt cttagaaaaa
     6301 ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt
     6361 ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc
     6421 aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt
     6481 cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg
     6541 tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc agccattacg
     6601 ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc
     6661 gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg
     6721 gcgcaggaac actgccagcg catcaacaat gttttcacct gaatcaggat attcttctaa
     6781 tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt
     6841 acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac
     6901 catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg
     6961 cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg
     7021 agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg gcctagagca
     7081 agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga
     7141 cagttttatt gttcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     7201 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     7261 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     7321 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     7381 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     7441 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     7501 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     7561 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     7621 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     7681 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     7741 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     7801 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     7861 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     7921 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     7981 tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     8041 tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     8101 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
     8161 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     8221 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
     8281 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
     8341 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
     8401 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
     8461 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
     8521 gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg
     8581 cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta
     8641 ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag
     8701 ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt
     8761 gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt
     8821 gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg
     8881 agcacgatca tgcgcacccg tggggccgcc atgccggcga taatggcctg cttctcgccg
     8941 aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat
     9001 accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg
     9061 acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt
     9121 gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
     9181 aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg
     9241 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     9301 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
     9361 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
     9421 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
     9481 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg
     9541 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
     9601 catcgcagtg ggaacgatac cctcattcag catttgcatg gtttgttgaa aaccggacat
     9661 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
     9721 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
     9781 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
     9841 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
     9901 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
     9961 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
    10021 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
    10081 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
    10141 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
    10201 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
    10261 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
    10321 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
    10381 ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca
    10441 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg
    10501 catg
//
