LOCUS       sequence-160091-        9411 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Bacterial expression - MBP-his tagged LbCpf1 (E.coli codon 
            optimized).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9411)
  AUTHORS   Kim D, Kim J, Hur JK, Been KW, Yoon SH, Kim JS
  TITLE     Genome-wide analysis reveals specificities of Cpf1 endonucleases in 
            human cells.
  JOURNAL   Nat Biotechnol. 2016 Jun 6. doi: 10.1038/nbt.3609.
  PUBMED    27272384
REFERENCE   2  (bases 1 to 9411)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2016 Jun 6. doi: 10.1038/nbt.3609."
COMMENT     Sequence Label: pMAL-his-LbCpf1-EC
FEATURES             Location/Qualifiers
     source          1..9411
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..1101
                     /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
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELVKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1075..1098
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1150..1161
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1174..1191
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1192..1212
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1231..1251
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1252..4935
                     /codon_start=1
                     /product="CRISPR-associated protein from Lachnospiraceae 
                     bacterium ND2006 (Zetsche et al., 2015)"
                     /label=LbCpf1
                     /translation="MSKLEKFTNCYSLSKTLRFKAIPVGKTQENIDNKRLLVEDEKRAE
                     DYKGVKKLLDRYYLSFINDVLHSIKLKNLNNYISLFRKKTRTEKENKELENLEINLRKE
                     IAKAFKGNEGYKSLFKKDIIETILPEFLDDKDEIALVNSFNGFTTAFTGFFDNRENMFS
                     EEAKSTSIAFRCINENLTRYISNMDIFEKVDAIFDKHEVQEIKEKILNSDYDVEDFFEG
                     EFFNFVLTQEGIDVYNAIIGGFVTESGEKIKGLNEYINLYNQKTKQKLPKFKPLYKQVL
                     SDRESLSFYGEGYTSDEEVLEVFRNTLNKNSEIFSSIKKLEKLFKNFDEYSSAGIFVKN
                     GPAISTISKDIFGEWNVIRDKWNAEYDDIHLKKKAVVTEKYEDDRRKSFKKIGSFSLEQ
                     LQEYADADLSVVEKLKEIIIQKVDEIYKVYGSSEKLFDADFVLEKSLKKNDAVVAIMKD
                     LLDSVKSFENYIKAFFGEGKETNRDESFYGDFVLAYDILLKVDHIYDAIRNYVTQKPYS
                     KDKFKLYFQNPQFMGGWDKDKETDYRATILRYGSKYYLAIMDKKYAKCLQKIDKDDVNG
                     NYEKINYKLLPGPNKMLPKVFFSKKWMAYYNPSEDIQKIYKNGTFKKGDMFNLNDCHKL
                     IDFFKDSISRYPKWSNAYDFNFSETEKYKDIAGFYREVEEQGYKVSFESASKKEVDKLV
                     EEGKLYMFQIYNKDFSDKSHGTPNLHTMYFKLLFDENNHGQIRLSGGAELFMRRASLKK
                     EELVVHPANSPIANKNPDNPKKTTTLSYDVYKDKRFSEDQYELHIPIAINKCPKNIFKI
                     NTEVRVLLKHDDNPYVIGIDRGERNLLYIVVVDGKGNIVEQYSLNEIINNFNGIRIKTD
                     YHSLLDKKEKERFEARQNWTSIENIKELKAGYISQVVHKICELVEKYDAVIALEDLNSG
                     FKNSRVKVEKQVYQKFEKMLIDKLNYMVDKKSNPCATGGALKGYQITNKFESFKSMSTQ
                     NGFIFYIPAWLTSKIDPSTGFVNLLKTKYTSIADSKKFISSFDRIMYVPEEDLFEFALD
                     YKNFSRTDADYIKKWKLYSYGNRIRIFRNPKKNNVFDWEEVCLTSAYKELFNKYGINYQ
                     QGDIRALLCEQSDKAFYSSFMALMSLMLQMRNSITGRTDVDFLISPVKNSDGIFYDSRN
                     YEAQENAILPKNADANGAYNIARKVLWAIGQFKKAEDEKLDKVKIAISNKEWLEYAQTS
                     VKH"
     terminator      4970..5056
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5148..5175
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        5203..5293
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5294..5362,5363..6154)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5512..5531)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6242..6830
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6731..6750
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6984..7001
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7184..7206
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(7200..7391)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        7887..7964
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             7965..9047
                     /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     complement(7984..8003)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(9290..9295,9296..9311,9312..9318)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    9326..9342
                     /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)."
     primer_bind     9331..9353
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 atgaaaatcg aagaaggtaa actggtaatc tggattaacg gcgataaagg ctataacggt
       61 ctcgctgaag tcggtaagaa attcgagaaa gataccggaa ttaaagtcac cgttgagcat
      121 ccggataaac tggaagagaa attcccacag gttgcggcaa ctggcgatgg ccctgacatt
      181 atcttctggg cacacgaccg ctttggtggc tacgctcaat ctggcctgtt ggctgaaatc
      241 accccggaca aagcgttcca ggacaagctg tatccgttta cctgggatgc cgtacgttac
      301 aacggcaagc tgattgctta cccgatcgct gttgaagcgt tatcgctgat ttataacaaa
      361 gatctgctgc cgaacccgcc aaaaacctgg gaagagatcc cggcgctgga taaagaactg
      421 aaagcgaaag gtaagagcgc gctgatgttc aacctgcaag aaccgtactt cacctggccg
      481 ctgattgctg ctgacggggg ttatgcgttc aagtatgaaa acggcaagta cgacattaaa
      541 gacgtgggcg tggataacgc tggcgcgaaa gcgggtctga ccttcctggt tgacctgatt
      601 aaaaacaaac acatgaatgc agacaccgat tactccatcg cagaagctgc ctttaataaa
      661 ggcgaaacag cgatgaccat caacggcccg tgggcatggt ccaacatcga caccagcaaa
      721 gtgaattatg gtgtaacggt actgccgacc ttcaagggtc aaccatccaa accgttcgtt
      781 ggcgtgctga gcgcaggtat taacgccgcc agtccgaaca aagagctggc aaaagagttc
      841 ctcgaaaact atctgctgac tgatgaaggt ctggaagcgg ttaataaaga caaaccgctg
      901 ggtgccgtag cgctgaagtc ttacgaggaa gagttggtga aagatccgcg tattgccgcc
      961 actatggaaa acgcccagaa aggtgaaatc atgccgaaca tcccgcagat gtccgctttc
     1021 tggtatgccg tgcgtactgc ggtgatcaac gccgccagcg gtcgtcagac tgtcgatgaa
     1081 gccctgaaag acgcgcagac taattcgagc tcgaacaaca acaacaataa caataacaac
     1141 aacctcggga tcgagggaag gatttcacat atgcatcatc atcatcatca tgagaacctg
     1201 tacttccagg gcgaattcgg ctccggacct ccaaagaaaa agagaaaagt aatgagcaaa
     1261 ctggaaaaat ttacgaattg ttatagcctg tccaagaccc tgcgtttcaa agccatcccc
     1321 gttggcaaaa cccaggagaa tattgataat aaacgtctgc tggttgagga tgaaaaaaga
     1381 gcagaagact ataagggagt caaaaaactg ctggatcggt actacctgag ctttataaat
     1441 gacgtgctgc atagcattaa actgaaaaat ctgaataact atattagtct gttccgcaag
     1501 aaaacccgaa cagagaaaga aaataaagag ctggaaaacc tggagatcaa tctgcgtaaa
     1561 gagatcgcaa aagcttttaa aggaaatgaa ggttataaaa gcctgttcaa aaaagacatt
     1621 attgaaacca tcctgccgga atttctggat gataaagacg agatagcgct cgtgaacagc
     1681 ttcaacgggt tcacgaccgc cttcacgggc tttttcgata acagggaaaa tatgttttca
     1741 gaggaagcca aaagcacctc gatagcgttc cgttgcatta atgaaaattt gacaagatat
     1801 atcagcaaca tggatatttt cgagaaagtt gatgcgatct ttgacaaaca tgaagtgcag
     1861 gagattaagg aaaaaattct gaacagcgat tatgatgttg aggatttttt cgagggggaa
     1921 ttttttaact ttgtactgac acaggaaggt atagatgtgt ataatgctat tatcggcggg
     1981 ttcgttaccg aatccggcga gaaaattaag ggtctgaatg agtacatcaa tctgtataac
     2041 caaaagacca aacagaaact gccaaaattc aaaccgctgt acaagcaagt cctgagcgat
     2101 cgggaaagct tgagctttta cggtgaaggt tataccagcg acgaggaggt actggaggtc
     2161 tttcgcaata ccctgaacaa gaacagcgaa attttcagct ccattaaaaa gctggagaaa
     2221 ctgtttaaga attttgacga gtacagcagc gcaggtattt ttgtgaagaa cggacctgcc
     2281 ataagcacca ttagcaagga tatttttgga gagtggaatg ttatccgtga taaatggaac
     2341 gcggaatatg atgacataca cctgaaaaag aaggctgtgg taactgagaa atatgaagac
     2401 gatcgccgca aaagctttaa aaaaatcggc agctttagcc tggagcagct gcaggaatat
     2461 gcggacgccg acctgagcgt ggtcgagaaa ctgaaggaaa ttattatcca aaaagtggat
     2521 gagatttaca aggtatatgg tagcagcgaa aaactgtttg atgcggactt cgttctggaa
     2581 aaaagcctga aaaaaaatga tgctgttgtt gcgatcatga aagacctgct cgatagcgtt
     2641 aagagctttg aaaattacat taaagcattc tttggcgagg gcaaagaaac aaacagagac
     2701 gaaagctttt atggcgactt cgtcctggct tatgacatcc tgttgaaggt agatcatata
     2761 tatgatgcaa ttcgtaatta cgtaacccaa aagccgtaca gcaaagataa gttcaaactg
     2821 tatttccaga acccgcagtt tatgggtggc tgggacaaag acaaggagac agactatcgc
     2881 gccactattc tgcgttacgg cagcaagtac tatctcgcca tcatggacaa aaaatatgca
     2941 aagtgtctgc agaaaatcga taaagacgac gtgaacggaa attacgaaaa gattaattat
     3001 aagctgctgc cagggcccaa caagatgtta ccgaaagtat ttttttccaa aaaatggatg
     3061 gcatactata acccgagcga ggatatacag aagatttaca aaaatgggac cttcaaaaag
     3121 ggggatatgt tcaatctgaa tgactgccac aaactgatcg atttttttaa agatagcatc
     3181 agccgttatc ctaaatggtc aaacgcgtat gattttaatt tctccgaaac ggagaaatat
     3241 aaagacattg ctggtttcta tcgcgaagtc gaagaacagg gttataaagt tagctttgaa
     3301 tcggccagca agaaagaggt tgataaactg gtggaggagg gtaagctgta tatgtttcag
     3361 atttataaca aagactttag cgacaaaagc cacggtactc ctaatctgca tacgatgtac
     3421 tttaaactgc tgtttgatga gaataaccac ggccaaatcc gtctctccgg tggagcagaa
     3481 ctttttatgc ggcgtgcgag cctaaaaaag gaagaactgg tggtgcatcc cgccaacagc
     3541 ccgattgcta acaaaaatcc agataatcct aagaagacca ccacactgtc gtacgatgtc
     3601 tataaggata aacgtttctc ggaagaccag tatgaattgc atataccgat agcaattaat
     3661 aaatgcccaa aaaacatttt caaaatcaac actgaagttc gtgtgctgct gaaacatgat
     3721 gataatccgt atgtgatcgg aattgaccgt ggggagagaa atctgctgta tattgtagtc
     3781 gttgatggca agggcaacat cgttgagcag tatagcctga atgaaataat taataatttt
     3841 aacggtatac gtattaaaac cgactatcat agcctgctgg ataaaaagga gaaagagcgt
     3901 tttgaggcac gccaaaattg gacgagcatc gaaaacatca aggaactgaa ggcaggatat
     3961 atcagccaag tagtccataa aatctgtgaa ctggtggaga agtacgacgc tgtcattgcc
     4021 ctggaagacc tcaatagcgg ctttaaaaac agccgggtga aggtggagaa acaggtatac
     4081 caaaagtttg aaaagatgct cattgataag ctgaactata tggttgataa aaagagcaac
     4141 ccgtgcgcca ctggcggtgc actgaaaggg taccaaatta ccaataaatt tgaaagcttt
     4201 aaaagcatga gcacgcagaa tgggtttatt ttttatatac cagcatggct gacgagcaag
     4261 attgacccca gcactggttt tgtcaatctg ctgaaaacca aatacacaag cattgcggat
     4321 agcaaaaaat ttatttcgag cttcgaccgt attatgtatg ttccggagga agatctgttt
     4381 gaatttgccc tggattataa aaacttcagc cgcaccgatg cagattatat caaaaaatgg
     4441 aagctgtaca gttatggtaa tcgtatacgt atcttccgta atccgaagaa aaacaatgtg
     4501 ttcgattggg aagaggtctg tctgaccagc gcgtataaag aactgttcaa caagtacgga
     4561 ataaattatc agcaaggtga cattcgcgca ctgctgtgtg aacagtcaga taaagcattt
     4621 tatagcagct ttatggcgct gatgagcctg atgctccaga tgcgcaacag cataaccggt
     4681 cgcacagatg ttgactttct gatcagccct gtgaagaata gcgacggcat cttctacgat
     4741 tccaggaact atgaagcaca ggaaaacgct attctgccta aaaatgccga tgccaacggc
     4801 gcctataata ttgcacggaa ggttctgtgg gcgattggac agttcaagaa agcggaagat
     4861 gagaagctgg ataaggtaaa aattgctatt agcaataagg aatggctgga gtacgcacag
     4921 acatcggtta aacacgcggc cgcttccctg caggtaatta aataagcttc aaataaaacg
     4981 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     5041 cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg
     5101 gtggcgggca ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct
     5161 gacggatggc ctttttgcgt ttctacaaac tctttcggtc cgttgtttat ttttctaaat
     5221 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     5281 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     5341 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     5401 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     5461 gagttttcgc cccgaagaac gtttcccaat gatgagcact tttaaagttc tgctatgtgg
     5521 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     5581 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     5641 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     5701 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     5761 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     5821 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     5881 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     5941 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     6001 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     6061 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     6121 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     6181 actttagatt gatttcctta ggactgagcg tcaaccccgt agaaaagatc aaaggatctt
     6241 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6301 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6361 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     6421 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6481 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6541 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6601 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6661 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6721 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6781 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6841 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     6901 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     6961 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     7021 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatataa ggtgcactgt
     7081 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     7141 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     7201 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg
     7261 tggtcgtgca gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc
     7321 tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc
     7381 tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg
     7441 atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg
     7501 gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact
     7561 cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag
     7621 catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga
     7681 ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg
     7741 cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg
     7801 caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggc
     7861 caggacccaa cgctgcccga aattccgaca ccatcgaatg gtgcaaaacc tttcgcggta
     7921 tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt
     7981 tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc
     8041 aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga
     8101 attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg
     8161 ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc
     8221 gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag
     8281 cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact
     8341 atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt
     8401 tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg
     8461 gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag
     8521 cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca
     8581 ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt
     8641 ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca
     8701 acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg
     8761 cggatatttc ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt
     8821 taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc
     8881 aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa
     8941 gaaaaaccac cctggcgccc aatacacaaa ccgcctctcc ccgcgcgttg gccgattcat
     9001 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     9061 aatgtaagtt agctcactca ttaggcacaa ttctcatgtt tgacagctta tcatcgactg
     9121 cacggtgcac caatgcttct ggcgtcaggc agccatcgga agctgtggta tggctgtgca
     9181 ggtcgtaaat cactgcataa ttcgtgtcgc tcaaggcgca ctcccgttct ggataatgtt
     9241 ttttgcgccg acatcataac ggttctggca aatattctga aatgagctgt tgacaattaa
     9301 tcatcggctc gtataatgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     9361 cagtccgttt aggtgttttc acgagcaatt gaccaacaag gaccatagat t
//
