LOCUS       sequence-320077-       10632 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression, protein expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10632)
  AUTHORS   Sreekanth V, Zhou Q, Kokkonda P, Bermudez-Cabrera HC, Lim D, Law BK,
            Holmes BR, Chaudhary SK, Pergu R, Leger BS, Walker JA, Gifford DK, 
            Sherwood RI, Choudhary A
  TITLE     Chemogenetic System Demonstrates That Cas9 Longevity Impacts Genome 
            Editing Outcomes.
  JOURNAL   ACS Cent Sci. 2020 Dec 23;6(12):2228-2237. doi: 
            10.1021/acscentsci.0c00129. Epub 2020 Nov 18.
  PUBMED    33376784
REFERENCE   2  (bases 1 to 10632)
  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"; doi: 
            "10.1021/acscentsci.0c00129"; journalName: "ACS Cent Sci"; date: 
            "2020-12-23- 23"; volume: "6"; issue: "12"; pages: "2228-2237"
COMMENT     Sequence Label: pET SUMO NL-FKBP-Cas9
FEATURES             Location/Qualifiers
     source          1..10632
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     883..902
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(922..999)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1205..1224
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1308..1327
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1308..1326
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1327..1351
                     /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)."
     CDS             1403..1420
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1433..1720
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated and modified)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /note="modified to prevent cleavage by eukaryotic SUMO 
                     proteases"
                     /translation="DSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLRR
                     LMEAFAKRQGKEMDSLRFLYDGIRIQADQAPEDLDMEDNDIIEAHREQIGG"
     CDS             1727..1747
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(1748..1750,1751..2071)
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=FKBP-V
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     CDS             2774..3094
                     /codon_start=1
                     /product="F36V mutant of FK506-binding protein FKBP12"
                     /label=FKBP-V
                     /note="binds synthetic ligands that do not bind wild-type 
                     FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             4019..6517
                     /codon_start=1
                     /product="C-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(C)
                     /translation="KPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEI
                     SGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTY
                     AHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIH
                     DDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPE
                     NIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYL
                     QNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEV
                     VKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQI
                     LDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVV
                     GTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITL
                     ANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESIL
                     PKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIME
                     RSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALP
                     SKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLD
                     KVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATL
                     IHQSITGLYETRIDLSQLGGD"
     CDS             6518..6565
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             6587..6610
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6611..6655
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             6671..6688
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6741..6759)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6755..6802
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      6839..7294
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6926..6945)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7136..7157
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7320..7424
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7425..7493,7494..8285)
                     /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(7643..7662)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8456..9044
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8945..8964
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9198..9215
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    9230..9369
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(9295..9314)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9455..9477
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(9471..9662)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(10471..10632,1..921))
                     /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 gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt
       61 ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat
      121 atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg
      181 atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg
      241 ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt
      301 gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg
      361 gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg
      421 cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag
      481 aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag
      541 cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt
      601 acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc
      661 ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt
      721 ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat
      781 gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct
      841 ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc
      901 gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca
      961 tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct
     1021 tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg
     1081 ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg
     1141 ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc
     1201 catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg
     1261 ccacgatgcg tccggcgtag aggatcgaga tctcgatccc gcgaaattaa tacgactcac
     1321 tataggggaa ttgtgagcgg ataacaattc ccctctagaa ataattttgg tttaacttta
     1381 agaaggagat atacatatgg gtcatcacca tcatcatcac gggtccctgc aggactcaga
     1441 agtcaatcaa gaagctaagc cagaggtcaa gccagaagtc aagcctgaga ctcacatcaa
     1501 tttaaaggtg tccgatggat cttcagagat cttcttcaag atcaaaaaga ccactccttt
     1561 aagaaggctg atggaagcgt tcgctaaaag acagggtaag gaaatggact ccttaagatt
     1621 cttgtacgac ggtattagaa ttcaagctga tcaggcccct gaagatttgg acatggagga
     1681 taacgatatt attgaggctc accgcgaaca gattggaggt gctagcgaaa atctgtattt
     1741 tcagggaatg ggagtgcagg tggaaaccat ctccccagga gacgggcgca ccttccccaa
     1801 gcgcggccag acctgcgtgg tgcactacac cgggatgctt gaagatggaa agaaagttga
     1861 ttcctcccgg gacagaaaca agccctttaa gtttatgcta ggcaagcagg aggtgatccg
     1921 aggctgggaa gaaggggttg cccagatgag tgtgggtcag agagccaaac tgactatatc
     1981 tccagattat gcctatggtg ccactgggca cccaggcatc atcccaccac atgccactct
     2041 cgtcttcgat gtggagcttc taaaactgga agacaagaag tacagcatcg gcctggacat
     2101 cggcaccaac tctgtgggct gggccgtgat caccgacgag tacaaggtgc ccagcaagaa
     2161 attcaaggtg ctgggcaaca ccgaccggca cagcatcaag aagaacctga tcggagccct
     2221 gctgttcgac agcggcgaaa cagccgaggc cacccggctg aagagaaccg ccagaagaag
     2281 atacaccaga cggaagaacc ggatctgcta tctgcaagag atcttcagca acgagatggc
     2341 caaggtggac gacagcttct tccacagact ggaagagtcc ttcctggtgg aagaggataa
     2401 gaagcacgag cggcacccca tcttcggcaa catcgtggac gaggtggcct accacgagaa
     2461 gtaccccacc atctaccacc tgagaaagaa actggtggac agcaccgaca aggccgacct
     2521 gcggctgatc tatctggccc tggcccacat gatcaagttc cggggccact tcctgatcga
     2581 gggcgacctg aaccccgaca acagcgacgt ggacaagctg ttcatccagc tggtgcagac
     2641 ctacaaccag ctgttcgagg aaaaccccat caacgccagc ggcgtggacg ccaaggccat
     2701 cctgtctgcc agactgagca agagcagacg gctggaaaat ctgatcgccc agctgcccgg
     2761 cggaagtggc agcggagtgc aggtggaaac catctcccca ggagacgggc gcaccttccc
     2821 caagcgcggc cagacctgcg tggtgcacta caccgggatg cttgaagatg gaaagaaagt
     2881 tgattcctcc cgggacagaa acaagccctt taagtttatg ctaggcaagc aggaggtgat
     2941 ccgaggctgg gaagaagggg ttgcccagat gagtgtgggt cagagagcca aactgactat
     3001 atctccagat tatgcctatg gtgccactgg gcacccaggc atcatcccac cacatgccac
     3061 tctcgtcttc gatgtggagc ttctaaaact ggaaggcagt ggaagcgaga agaagaatgg
     3121 cctgttcgga aacctgattg ccctgagcct gggcctgacc cccaacttca agagcaactt
     3181 cgacctggcc gaggatgcca aactgcagct gagcaaggac acctacgacg acgacctgga
     3241 caacctgctg gcccagatcg gcgaccagta cgccgacctg tttctggccg ccaagaacct
     3301 gtccgacgcc atcctgctga gcgacatcct gagagtgaac accgagatca ccaaggcccc
     3361 cctgagcgcc tctatgatca agagatacga cgagcaccac caggacctga ccctgctgaa
     3421 agctctcgtg cggcagcagc tgcctgagaa gtacaaagag attttcttcg accagagcaa
     3481 gaacggctac gccggctaca ttgacggcgg agccagccag gaagagttct acaagttcat
     3541 caagcccatc ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc tgaacagaga
     3601 ggacctgctg cggaagcagc ggaccttcga caacggcagc atcccccacc agatccacct
     3661 gggagagctg cacgccattc tgcggcggca ggaagatttt tacccattcc tgaaggacaa
     3721 ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc tactacgtgg gccctctggc
     3781 caggggaaac agcagattcg cctggatgac cagaaagagc gaggaaacca tcaccccctg
     3841 gaacttcgag gaagtggtgg acaagggcgc ttccgcccag agcttcatcg agcggatgac
     3901 caacttcgat aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga
     3961 gtacttcacc gtgtataacg agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa
     4021 gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa
     4081 ccggaaagtg accgtgaagc agctgaaaga ggactacttc aagaaaatcg agtgcttcga
     4141 ctccgtggaa atctccggcg tggaagatcg gttcaacgcc tccctgggca cataccacga
     4201 tctgctgaaa attatcaagg acaaggactt cctggacaat gaggaaaacg aggacattct
     4261 ggaagatatc gtgctgaccc tgacactgtt tgaggacaga gagatgatcg aggaacggct
     4321 gaaaacctat gcccacctgt tcgacgacaa agtgatgaag cagctgaagc ggcggagata
     4381 caccggctgg ggcaggctga gccggaagct gatcaacggc atccgggaca agcagtccgg
     4441 caagacaatc ctggatttcc tgaagtccga cggcttcgcc aacagaaact tcatgcagct
     4501 gatccacgac gacagcctga cctttaaaga ggacatccag aaagcccagg tgtccggcca
     4561 gggcgatagc ctgcacgagc acattgccaa tctggccggc agccccgcca ttaagaaggg
     4621 catcctgcag acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc
     4681 cgagaacatc gtgatcgaaa tggccagaga gaaccagacc acccagaagg gacagaagaa
     4741 cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa gagctgggca gccagatcct
     4801 gaaagaacac cccgtggaaa acacccagct gcagaacgag aagctgtacc tgtactacct
     4861 gcagaatggg cgggatatgt acgtggacca ggaactggac atcaaccggc tgtccgacta
     4921 cgatgtggac catatcgtgc ctcagagctt tctgaaggac gactccatcg acaacaaggt
     4981 gctgaccaga agcgacaaga accggggcaa gagcgacaac gtgccctccg aagaggtcgt
     5041 gaagaagatg aagaactact ggcggcagct gctgaacgcc aagctgatta cccagagaaa
     5101 gttcgacaat ctgaccaagg ccgagagagg cggcctgagc gaactggata aggccggctt
     5161 catcaagaga cagctggtgg aaacccggca gatcacaaag cacgtggcac agatcctgga
     5221 ctcccggatg aacactaagt acgacgagaa tgacaagctg atccgggaag tgaaagtgat
     5281 caccctgaag tccaagctgg tgtccgattt ccggaaggat ttccagtttt acaaagtgcg
     5341 cgagatcaac aactaccacc acgcccacga cgcctacctg aacgccgtcg tgggaaccgc
     5401 cctgatcaaa aagtacccta agctggaaag cgagttcgtg tacggcgact acaaggtgta
     5461 cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta
     5521 cttcttctac agcaacatca tgaacttttt caagaccgag attaccctgg ccaacggcga
     5581 gatccggaag cggcctctga tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa
     5641 gggccgggat tttgccaccg tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa
     5701 aaagaccgag gtgcagacag gcggcttcag caaagagtct atcctgccca agaggaacag
     5761 cgataagctg atcgccagaa agaaggactg ggaccctaag aagtacggcg gcttcgacag
     5821 ccccaccgtg gcctattctg tgctggtggt ggccaaagtg gaaaagggca agtccaagaa
     5881 actgaagagt gtgaaagagc tgctggggat caccatcatg gaaagaagca gcttcgagaa
     5941 gaatcccatc gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat
     6001 caagctgcct aagtactccc tgttcgagct ggaaaacggc cggaagagaa tgctggcctc
     6061 tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc tccaaatatg tgaacttcct
     6121 gtacctggcc agccactatg agaagctgaa gggctccccc gaggataatg agcagaaaca
     6181 gctgtttgtg gaacagcaca agcactacct ggacgagatc atcgagcaga tcagcgagtt
     6241 ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa
     6301 gcaccgggat aagcccatca gagagcaggc cgagaatatc atccacctgt ttaccctgac
     6361 caatctggga gcccctgccg ccttcaagta ctttgacacc accatcgacc ggaagaggta
     6421 caccagcacc aaagaggtgc tggacgccac cctgatccac cagagcatca ccggcctgta
     6481 cgagacacgg atcgacctgt ctcagctggg aggcgacaaa aggccggcgg ccacgaaaaa
     6541 ggccggccag gcaaaaaaga aaaagtaact cgagggtggt ggtagcgatt ataaagatga
     6601 tgatgataaa ggcctgaacg atatctttga agcccagaaa attgaatggc acgagtaaaa
     6661 gcttgtcgag caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga
     6721 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     6781 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac
     6841 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     6901 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     6961 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     7021 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     7081 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     7141 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     7201 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     7261 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     7321 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     7381 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     7441 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     7501 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     7561 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     7621 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     7681 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     7741 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     7801 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     7861 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     7921 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa
     7981 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     8041 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     8101 gctggtttat tgctgataaa tctggagccg gtgagcgtgg tagtcgcggt atcattgcag
     8161 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     8221 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     8281 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     8341 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     8401 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     8461 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     8521 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     8581 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     8641 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     8701 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     8761 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     8821 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     8881 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     8941 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     9001 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     9061 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     9121 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     9181 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     9241 attttctcct tacgcatctg tgcggtattt cacaccgcaa tggtgcactc tcagtacaat
     9301 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
     9361 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     9421 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     9481 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga
     9541 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
     9601 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
     9661 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga
     9721 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt
     9781 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa
     9841 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg
     9901 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa
     9961 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag
    10021 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc
    10081 cagcctagcc gggtcctcaa cgacaggagc acgatcatgc gcacccgtgg ggccgccatg
    10141 ccggcgataa tggcctgctt ctcgccgaaa cgtttggtgg cgggaccagt gacgaaggct
    10201 tgagcgaggg cgtgcaagat tccgaatacc gcaagcgaca ggccgatcat cgtcgcgctc
    10261 cagcgaaagc ggtcctcgcc gaaaatgacc cagagcgctg ccggcacctg tcctacgagt
    10321 tgcatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg
    10381 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga
    10441 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
    10501 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
    10561 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
    10621 ctggccctga ga
//
