LOCUS       sequence-346134-       10162 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 6xHis-MBP tagged PrCas12a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10162)
  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 10162)
  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-huPrCas12a
FEATURES             Location/Qualifiers
     source          1..10162
                     /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             5117..5137
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5144..5233
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             5234..5251
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5304..5322)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5318..5365
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      5402..5857
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5489..5508)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5699..5720
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5950..6765)
                     /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(5950..6765)
                     /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     6673..6692
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6757..6776
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6887..7475
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7376..7395
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7629..7646
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    7661..7800
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(7726..7745)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7886..7908
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(7902..8093)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(8902..9984)
                     /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     9946..9965
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(9985..10062)
                     /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 agcctgctga agaaccacga gaacttcccc aagatcggca acaaggagat
     1861 catcaagctg ctgcagagcc tgtacgacaa cgacaccgac tacaaggccc tggacagctt
     1921 cagcaacttc tacacctact tcagcagcta caacgaggtg cgcaagaacc tgtacagcga
     1981 cgaggagaag agcagcaccg tggcctaccg cctgatcaac gagaacctgc ccaagttcct
     2041 ggacaacatc aaggcctacg ccatcgccaa gaaggccggc gtgcgcgccg agggcctgag
     2101 cgaggaggac caggactgcc tgttcatcat cgagaccttc gagcgcaccc tgacccagga
     2161 cggcatcgac aactacaacg ccgccatcgg caagctgaac accgccatca acctgttcaa
     2221 ccagcagaac aagaagcagg agggcttccg caaggtgccc cagatgaagt gcctgtacaa
     2281 gcagatcctg agcgaccgcg aggaggcctt catcgacgag ttcagcgacg acgaggacct
     2341 gatcaccaac atcgagagct tcgccgagaa catgaacgtg ttcctgaaca gcgagatcat
     2401 caccgacttc aagatcgccc tggtggagag cgacggcagc ctggtgtaca tcaagaacga
     2461 cgtgagcaag accagcttca gcaacatcgt gttcggcagc tggaacgcca tcgacgagaa
     2521 gctgagcgac gagtacgacc tggccaacag caagaagaag aaggacgaga agtactacga
     2581 gaagcgccag aaggagctga agaagaacaa gagctacgac ctggagacca tcatcggcct
     2641 gttcgacgac aacagcgacg 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 gaacttcaac cgcgccaccc tgctgaacgg ctgggacaag aacaaggaga ccgacaacct
     3061 gggcatcctg ttcgagaagg acggcatgta ctacctgggc atcatgaaca ccaaggccaa
     3121 caagatcttc gtgaacatcc ccaaggccac cagcaacgac gtgtaccaca aggtgaacta
     3181 caagctgctg cccggcccca acaagatgct gcccaaggtg ttcttcgccc agagcaacct
     3241 ggactactac aagcccagcg aggagctgct ggccaagtac aaggccggca cccacaagaa
     3301 gggcgacaac ttcagcctgg aggactgcca cgccctgatc gacttcttca aggccagcat
     3361 cgagaagcac cccgactgga gcagcttcgg cttcgagttc agcgagacct gcacctacga
     3421 ggacctgagc ggcttctacc gcgaggtgga gaagcagggc tacaagatca cctacaccga
     3481 cgtggacgcc gactacatca ccagcctggt ggagcgcgac gagctgtacc tgttccagat
     3541 ctacaacaag gacttcagcc cctacagcaa gggcaacctg aacctgcaca ccatctacct
     3601 gcagatgctg ttcgaccagc gcaacctgaa caacgtggtg tacaagctga acggcgaggc
     3661 cgaggtgttc taccgccccg ccagcatcaa cgacgaggag gtgatcatcc acaaggccgg
     3721 cgaggagatc aagaacaaga acagcaagcg cgccgtggac aagcccacca gcaagttcgg
     3781 ctacgacatc atcaaggacc gccgctacag caaggacaag ttcatgctgc acatccccgt
     3841 gaccatgaac ttcggcgtgg acgagacccg ccgcttcaac gacgtggtga acgacgccct
     3901 gcgcaacgac gagaaggtgc gcgtgatcgg catcgaccgc ggcgagcgca acctgctgta
     3961 cgtggtggtg gtggacaccg acggcaccat cctggagcag atcagcctga acagcatcat
     4021 caacaacgag tacagcatcg agaccgacta ccacaagctg ctggacgaga aggagggcga
     4081 ccgcgaccgc gcccgcaaga actggaccac catcgagaac atcaaggagc tgaaggaggg
     4141 ctacctgagc caggtggtga acgtgatcgc caagctggtg ctgaagtaca acgccatcat
     4201 ctgcctggag gacctgaact tcggcttcaa gcgcggccgc cagaaggtgg agaagcaggt
     4261 gtaccagaag ttcgagaaga tgctgatcga caagctgaac tacctggtga tcgacaagag
     4321 ccgcaagcag gacaagcccg aggagttcgg cggcgccctg aacgccctgc agctgaccag
     4381 caagttcacc agcttcaagg acatgggcaa gcagaccggc atcatctact acgtgcccgc
     4441 ctacctgacc agcaagatcg accccaccac cggcttcgcc aacctgttct acgtgaagta
     4501 cgagaacgtg gagaaggcca aggagttctt cagccgcttc gacagcatca gctacaacaa
     4561 cgagagcggc tacttcgagt tcgccttcga ctacaagaag ttcaccgacc gcgcctgcgg
     4621 cgcccgcagc cagtggaccg tgtgcaccta cggcgagcgc atcatcaagt tccgcaacac
     4681 cgagaagaac aacagcttcg acgacaagac catcgtgctg agcgaggagt tcaaggagct
     4741 gttcagcatc tacggcatca gctacgagga cggcgccgag ctgaagaaca agatcatgag
     4801 cgtggacgag gccgacttct tccgcagcct gacccgcctg ttccagcaga ccatgcagat
     4861 gcgcaacagc agcaacgacg tgacccgcga ctacatcatc agccccatca tgaacgaccg
     4921 cggcgagttc ttcaacagcg aggcctgcga cgccagcaag cccaaggacg ccgacgccaa
     4981 cggcgccttc aacatcgccc gcaagggcct gtgggtgctg gagcagatcc gcaacacccc
     5041 cagcggcgac aagctgaacc tggccatgag caacgccgag tggctggagt acgcccagcg
     5101 caaccagatc gctagcccaa agaagaagag gaaagtctct agataccctt atgatgttcc
     5161 agattatgcc ggatacccat acgatgtccc tgactatgca ggctcctacc cttatgacgt
     5221 cccagactac gcgcaccacc accaccacca ctgagatccg gctgctaaca aagcccgaaa
     5281 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
     5341 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat tggcgaatgg
     5401 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     5461 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     5521 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     5581 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     5641 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     5701 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     5761 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     5821 aacgcgaatt ttaacaaact agtaacgttt acaatttcag gtggcacttt tcggggaaat
     5881 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     5941 aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg
     6001 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
     6061 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
     6121 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
     6181 agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc
     6241 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
     6301 tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac
     6361 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatgt tttcacctga
     6421 atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa
     6481 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
     6541 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
     6601 tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga
     6661 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
     6721 taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt
     6781 actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa cgtgagtttt
     6841 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     6901 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     6961 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     7021 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     7081 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     7141 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     7201 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     7261 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     7321 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     7381 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     7441 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     7501 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     7561 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     7621 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     7681 ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga
     7741 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
     7801 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     7861 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     7921 tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca
     7981 cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc
     8041 tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc
     8101 tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg
     8161 ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa
     8221 caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc
     8281 ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc
     8341 cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa
     8401 ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac
     8461 gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc
     8521 cgggtcctca acgacaggag cacgatcatg cgcacccgtg gggccgccat gccggcgata
     8581 atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg
     8641 gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag
     8701 cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag ttgcatgata
     8761 aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg
     8821 actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa
     8881 cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     8941 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt
     9001 ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg
     9061 agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat
     9121 ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga
     9181 gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat
     9241 ctgatcgttg gcaaccagca tcgcagtggg aacgataccc tcattcagca tttgcatggt
     9301 ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg
     9361 attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa
     9421 tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag
     9481 tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc
     9541 aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc
     9601 cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc
     9661 tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg
     9721 atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga
     9781 ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg
     9841 aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg
     9901 gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac
     9961 atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta
    10021 tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct cgacgctctc
    10081 ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg ttgagcaccg
    10141 ccgccgcaag gaatggtgca tg
//
