LOCUS       sequence-237659-       10551 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression plasmid for 2xNLS FnCpf1 in pET-21a backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10551)
  AUTHORS   Liu P, Luk K, Shin M, Idrizi F, Kwok S, Roscoe B, Mintzer E, Suresh 
            S, Morrison K, Frazao JB, Bolukbasi MF, Ponnienselvan K, Luban J, 
            Zhu LJ, Lawson ND, Wolfe SA
  TITLE     Enhanced Cas12a editing in mammalian cells and zebrafish.
  JOURNAL   Nucleic Acids Res. 2019 Mar 20. pii: 5403491. doi: 
            10.1093/nar/gkz184.
  PUBMED    30892626
REFERENCE   2  (bases 1 to 10551)
  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: "Nucleic 
            Acids Res. 2019 Mar 20. pii: 5403491. doi: 10.1093/nar/gkz184."
COMMENT     Sequence Label: pET-21a_2xNLS_FnCpf1_MBP_protein_expression
FEATURES             Location/Qualifiers
     source          1..10551
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     660..679
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(699..776)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     982..1001
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1085..1104
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1085..1103
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1104..1128
                     /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             1143..1165
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5073..5120
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5127..5207
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             5208..5228
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5235..5255
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             5256..6356
                     /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     6330..6353
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             6363..6380
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6433..6451)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6447..6494
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      6531..6986
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6618..6637)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6828..6849
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7013..7117
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7118..7186,7187..7978)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7336..7355)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8149..8737
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8638..8657
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8891..8908
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    8923..9065
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(8991..9010)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9151..9173
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(9167..9358)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(10167..10551,1..698))
                     /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"
ORIGIN
        1 agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct
       61 atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc
      121 gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga
      181 tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc
      241 tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg
      301 gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga
      361 ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg
      421 ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc
      481 agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt
      541 gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt
      601 ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg
      661 gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc
      721 tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg
      781 atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag
      841 gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc
      901 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
      961 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
     1021 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg
     1081 aaattaatac gactcactat aggggaattg tgagcggata acaattcccc tctagaaata
     1141 attttgttta actttaagaa ggagatatac atatgagcat ctaccaggag ttcgtcaaca
     1201 agtattcact gagtaagaca ctgcggttcg agctgatccc acagggcaag acactggaga
     1261 acatcaaggc ccgaggcctg attctggacg atgagaagcg ggcaaaagac tataagaaag
     1321 ccaagcagat cattgataaa taccaccagt tctttatcga ggaaattctg agctccgtgt
     1381 gcatcagtga ggatctgctg cagaattact cagacgtgta cttcaagctg aagaagagcg
     1441 acgatgacaa cctgcagaag gacttcaagt ccgccaagga caccatcaag aaacagatta
     1501 gcgagtacat caaggactcc gaaaagttta aaaatctgtt caaccagaat ctgatcgatg
     1561 ctaagaaagg ccaggagtcc gacctgatcc tgtggctgaa acagtctaag gacaatggga
     1621 ttgaactgtt caaggctaac tccgatatca ctgatattga cgaggcactg gaaatcatca
     1681 agagcttcaa gggatggacc acatacttta aaggcttcca cgagaaccgc aagaacgtgt
     1741 actccagcaa cgacattcct acctccatca tctaccgaat cgtcgatgac aatctgccaa
     1801 agttcctgga gaacaaggcc aaatatgaat ctctgaagga caaagctccc gaggcaatta
     1861 attacgaaca gatcaagaaa gatctggctg aggaactgac attcgatatc gactataaga
     1921 ctagcgaggt gaaccagagg gtcttttccc tggacgaggt gtttgaaatc gccaatttca
     1981 acaattacct gaaccagtcc ggcattacta aattcaatac catcattggc gggaagtttg
     2041 tgaacgggga gaataccaag cgcaagggaa ttaacgaata catcaatctg tatagccagc
     2101 agatcaacga caaaactctg aagaaataca agatgtctgt gctgttcaaa cagatcctga
     2161 gtgataccga gtccaagtct tttgtcattg ataaactgga agatgactca gacgtggtca
     2221 ctaccatgca gagcttttat gagcagatcg ccgctttcaa gacagtggag gaaaaatcta
     2281 ttaaggaaac tctgagtctg ctgttcgatg acctgaaagc ccagaagctg gacctgagta
     2341 agatctactt caaaaacgat aagagtctga cagacctgtc acagcaggtg tttgatgact
     2401 attccgtgat tgggaccgcc gtcctggagt acattacaca gcagatcgct ccaaagaacc
     2461 tggataatcc ctctaagaaa gagcaggaac tgatcgctaa gaaaaccgag aaggcaaaat
     2521 atctgagtct ggaaacaatt aagctggcac tggaggagtt caacaagcac agggatattg
     2581 acaaacagtg ccgctttgag gaaatcctgg ccaacttcgc agccatcccc atgatttttg
     2641 atgagatcgc ccagaacaaa gacaatctgg ctcagatcag tattaagtac cagaaccagg
     2701 gcaagaaaga cctgctgcag gcttcagcag aagatgacgt gaaagccatc aaggatctgc
     2761 tggaccagac caacaatctg ctgcacaagc tgaaaatctt ccatattagt cagtcagagg
     2821 ataaggctaa tatcctggat aaagacgaac acttctacct ggtgttcgag gaatgttact
     2881 tcgagctggc aaacattgtc cccctgtata acaagattag gaactacatc acacagaagc
     2941 cttactctga cgagaagttt aaactgaact tcgaaaatag taccctggcc aacgggtggg
     3001 ataagaacaa ggagcctgac aacacagcta tcctgttcat caaggatgac aagtactatc
     3061 tgggagtgat gaataagaaa aacaataaga tcttcgatga caaagccatt aaggagaaca
     3121 aaggggaagg atacaagaaa atcgtgtata agctgctgcc cggcgcaaat aagatgctgc
     3181 ctaaggtgtt cttcagcgcc aagagtatca aattctacaa cccatccgag gacatcctgc
     3241 ggattagaaa tcactcaaca catactaaga acgggagccc ccagaaggga tatgagaaat
     3301 ttgagttcaa catcgaggat tgcaggaagt ttattgactt ctacaagcag agcatctcca
     3361 aacaccctga atggaaggat tttggcttcc ggttttccga cacacagaga tataactcta
     3421 tcgacgagtt ctaccgcgag gtggaaaatc aggggtataa gctgactttt gagaacattt
     3481 ctgaaagtta catcgacagc gtggtcaatc agggaaagct gtacctgttc cagatctata
     3541 acaaagattt ttcagcatac agcaagggca gaccaaacct gcatacactg tactggaagg
     3601 ccctgttcga tgagaggaat ctgcaggacg tggtctataa actgaacgga gaggccgaac
     3661 tgttttaccg gaagcagtct attcctaaga aaatcactca cccagctaag gaggccatcg
     3721 ctaacaagaa caaggacaat cctaagaaag agagcgtgtt cgaatacgat ctgattaagg
     3781 acaagcggtt caccgaagat aagttctttt tccattgtcc aatcaccatt aacttcaagt
     3841 caagcggcgc taacaagttc aacgacgaga tcaatctgct gctgaaggaa aaagcaaacg
     3901 atgtgcacat cctgagcatt gaccgaggag agcggcatct ggcctactat accctggtgg
     3961 atggcaaagg gaatatcatt aagcaggata cattcaacat cattggcaat gaccggatga
     4021 aaaccaacta ccacgataaa ctggctgcaa tcgagaagga tagagactca gctaggaagg
     4081 actggaagaa aatcaacaac attaaggaga tgaaggaagg ctatctgagc caggtggtcc
     4141 atgagattgc aaagctggtc atcgaataca atgccattgt ggtgttcgag gatctgaact
     4201 tcggctttaa gagggggcgc tttaaggtgg aaaaacaggt ctatcagaag ctggagaaaa
     4261 tgctgatcga aaagctgaat tacctggtgt ttaaagataa cgagttcgac aagaccggag
     4321 gcgtcctgag agcctaccag ctgacagctc cctttgaaac tttcaagaaa atgggaaaac
     4381 agacaggcat catctactat gtgccagccg gattcacttc caagatctgc cccgtgaccg
     4441 gctttgtcaa ccagctgtac cctaaatatg agtcagtgag caagtcccag gaatttttca
     4501 gcaagttcga taagatctgt tataatctgg acaaggggta cttcgagttt tccttcgatt
     4561 acaagaactt cggcgacaag gccgctaagg ggaaatggac cattgcctcc ttcggatctc
     4621 gcctgatcaa ctttcgaaat tccgataaaa accacaattg ggacactagg gaggtgtacc
     4681 caaccaagga gctggaaaag ctgctgaaag actactctat cgagtatgga catggcgaat
     4741 gcatcaaggc agccatctgt ggcgagagtg ataagaaatt tttcgccaag ctgacctcag
     4801 tgctgaatac aatcctgcag atgcggaact caaagaccgg gacagaactg gactatctga
     4861 ttagccccgt ggctgatgtc aacggaaact tcttcgacag cagacaggca cccaaaaata
     4921 tgcctcagga tgcagacgcc aacggggcct accacatcgg gctgaaggga ctgatgctgc
     4981 tgggccggat caagaacaat caggagggga agaagctgaa cctggtcatt aagaacgagg
     5041 aatacttcga gtttgtccag aatagaaata acaaaaggcc ggcggccacg aaaaaggccg
     5101 gccaggcaaa aaagaaaaag ggatcctacc catacgatgt tccagattac gcttatccct
     5161 acgacgtgcc tgattatgca tacccatatg atgtccccga ctatgcccca aagaagaagc
     5221 ggaaggtcaa gcttgaaaac ctgtacttcc aatccatgaa aatcgaagaa ggtaaactgg
     5281 taatctggat taacggcgat aaaggctata acggtctcgc tgaagtcggt aagaaattcg
     5341 agaaagatac cggaattaaa gtcaccgttg agcatccgga taaactggaa gagaaattcc
     5401 cacaggttgc ggcaactggc gatggccctg acattatctt ctgggcacac gaccgctttg
     5461 gtggctacgc tcaatctggc ctgttggctg aaatcacccc ggacaaagcg ttccaggaca
     5521 agctgtatcc gtttacctgg gatgccgtac gttacaacgg caagctgatt gcttacccga
     5581 tcgctgttga agcgttatcg ctgatttata acaaagatct gctgccgaac ccgccaaaaa
     5641 cctgggaaga gatcccggcg ctggataaag aactgaaagc gaaaggtaag agcgcgctga
     5701 tgttcaacct gcaagaaccg tacttcacct ggccgctgat tgctgctgac gggggttatg
     5761 cgttcaagta tgaaaacggc aagtacgaca ttaaagacgt gggcgtggat aacgctggcg
     5821 cgaaagcggg tctgaccttc ctggttgacc tgattaaaaa caaacacatg aatgcagaca
     5881 ccgattactc catcgcagaa gctgccttta ataaaggcga aacagcgatg accatcaacg
     5941 gcccgtgggc atggtccaac atcgacacca gcaaagtgaa ttatggtgta acggtactgc
     6001 cgaccttcaa gggtcaacca tccaaaccgt tcgttggcgt gctgagcgca ggtattaacg
     6061 ccgccagtcc gaacaaagag ctggcaaaag agttcctcga aaactatctg ctgactgatg
     6121 aaggtctgga agcggttaat aaagacaaac cgctgggtgc cgtagcgctg aagtcttacg
     6181 aggaagagtt ggcgaaagat ccacgtattg ccgccactat ggaaaacgcc cagaaaggtg
     6241 aaatcatgcc gaacatcccg cagatgtccg ctttctggta tgccgtgcgt actgcggtga
     6301 tcaacgccgc cagcggtcgt cagactgtcg atgaagccct gaaagacgcg cagactctcg
     6361 agcaccacca ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt
     6421 tggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
     6481 tgaggggttt tttgctgaaa ggaggaacta tatccggatt ggcgaatggg acgcgccctg
     6541 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     6601 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     6661 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     6721 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     6781 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     6841 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     6901 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     6961 taacaaaata ttaacgttta caatttcagg tggcactttt cggggaaatg tgcgcggaac
     7021 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     7081 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     7141 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     7201 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     7261 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     7321 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     7381 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     7441 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     7501 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     7561 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     7621 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt
     7681 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     7741 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     7801 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     7861 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     7921 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     7981 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     8041 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     8101 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     8161 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     8221 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     8281 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     8341 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     8401 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     8461 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     8521 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     8581 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     8641 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     8701 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     8761 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     8821 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     8881 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     8941 ccttacgcat ctgtgcggta tttcacaccg catatatggt gcactctcag tacaatctgc
     9001 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     9061 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     9121 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     9181 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     9241 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     9301 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg
     9361 atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga
     9421 ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg
     9481 gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc
     9541 agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc
     9601 agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac
     9661 ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc
     9721 ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc
     9781 ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggggcc gccatgccgg
     9841 cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag
     9901 cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc
     9961 gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca
    10021 tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg
    10081 agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga
    10141 gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
    10201 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc
    10261 agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg
    10321 ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt
    10381 ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact
    10441 accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc
    10501 gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt c
//
