LOCUS       sequence-315280-       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"
     rep_origin      532..1120
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1021..1040
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1274..1291
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1306..1445
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1371..1390)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1531..1553
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1547..1738)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2547..3629)
                     /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     3591..3610
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3630..3707)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3913..3932
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4016..4035
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4016..4034
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4035..4059
                     /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             4111..4128
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4141..4428
                     /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             4435..4455
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(4456..4458,4459..4779)
                     /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             5482..5802
                     /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             6727..9225
                     /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             9226..9273
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             9295..9318
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             9319..9363
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             9379..9396
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(9449..9467)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      9463..9510
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      9547..10002
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9634..9653)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9844..9865
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        10028..10132
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(10133..10201,10202..10632,1..361)
                     /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(10351..10370)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac
       61 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
      121 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
      181 gtttattgct gataaatctg gagccggtga gcgtggtagt cgcggtatca ttgcagcact
      241 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
      301 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
      361 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
      421 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
      481 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
      541 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
      601 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
      661 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
      721 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
      781 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
      841 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
      901 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
      961 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     1021 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     1081 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     1141 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     1201 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     1261 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     1321 tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc
     1381 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     1441 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     1501 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     1561 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     1621 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     1681 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg
     1741 atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga
     1801 ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg
     1861 gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc
     1921 agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc
     1981 agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac
     2041 ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc
     2101 ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc
     2161 ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggggcc gccatgccgg
     2221 cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag
     2281 cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc
     2341 gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca
     2401 tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg
     2461 agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga
     2521 gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     2581 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc
     2641 agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg
     2701 ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt
     2761 ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact
     2821 accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc
     2881 gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc
     2941 atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga
     3001 atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa
     3061 cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg
     3121 cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag
     3181 acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg
     3241 tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc
     3301 gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc
     3361 agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga
     3421 ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg
     3481 ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa
     3541 acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct
     3601 gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg
     3661 cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg
     3721 ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag
     3781 caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac
     3841 ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg
     3901 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
     3961 gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc gaaattaata
     4021 cgactcacta taggggaatt gtgagcggat aacaattccc ctctagaaat aattttggtt
     4081 taactttaag aaggagatat acatatgggt catcaccatc atcatcacgg gtccctgcag
     4141 gactcagaag tcaatcaaga agctaagcca gaggtcaagc cagaagtcaa gcctgagact
     4201 cacatcaatt taaaggtgtc cgatggatct tcagagatct tcttcaagat caaaaagacc
     4261 actcctttaa gaaggctgat ggaagcgttc gctaaaagac agggtaagga aatggactcc
     4321 ttaagattct tgtacgacgg tattagaatt caagctgatc aggcccctga agatttggac
     4381 atggaggata acgatattat tgaggctcac cgcgaacaga ttggaggtgc tagcgaaaat
     4441 ctgtattttc agggaatggg agtgcaggtg gaaaccatct ccccaggaga cgggcgcacc
     4501 ttccccaagc gcggccagac ctgcgtggtg cactacaccg ggatgcttga agatggaaag
     4561 aaagttgatt cctcccggga cagaaacaag ccctttaagt ttatgctagg caagcaggag
     4621 gtgatccgag gctgggaaga aggggttgcc cagatgagtg tgggtcagag agccaaactg
     4681 actatatctc cagattatgc ctatggtgcc actgggcacc caggcatcat cccaccacat
     4741 gccactctcg tcttcgatgt ggagcttcta aaactggaag acaagaagta cagcatcggc
     4801 ctggacatcg gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc
     4861 agcaagaaat tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc
     4921 ggagccctgc tgttcgacag cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc
     4981 agaagaagat acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac
     5041 gagatggcca aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa
     5101 gaggataaga agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac
     5161 cacgagaagt accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag
     5221 gccgacctgc ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc
     5281 ctgatcgagg gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg
     5341 gtgcagacct acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc
     5401 aaggccatcc tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag
     5461 ctgcccggcg gaagtggcag cggagtgcag gtggaaacca tctccccagg agacgggcgc
     5521 accttcccca agcgcggcca gacctgcgtg gtgcactaca ccgggatgct tgaagatgga
     5581 aagaaagttg attcctcccg ggacagaaac aagcccttta agtttatgct aggcaagcag
     5641 gaggtgatcc gaggctggga agaaggggtt gcccagatga gtgtgggtca gagagccaaa
     5701 ctgactatat ctccagatta tgcctatggt gccactgggc acccaggcat catcccacca
     5761 catgccactc tcgtcttcga tgtggagctt ctaaaactgg aaggcagtgg aagcgagaag
     5821 aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag
     5881 agcaacttcg acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac
     5941 gacctggaca acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc
     6001 aagaacctgt ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc
     6061 aaggcccccc tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc
     6121 ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac
     6181 cagagcaaga acggctacgc cggctacatt gacggcggag ccagccagga agagttctac
     6241 aagttcatca agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg
     6301 aacagagagg acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag
     6361 atccacctgg gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg
     6421 aaggacaacc gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc
     6481 cctctggcca ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc
     6541 accccctgga acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag
     6601 cggatgacca acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg
     6661 ctgtacgagt acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga
     6721 atgagaaagc ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc
     6781 aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag
     6841 tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca
     6901 taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag
     6961 gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag
     7021 gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg
     7081 cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag
     7141 cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc
     7201 atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg
     7261 tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt
     7321 aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg
     7381 cacaagcccg agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga
     7441 cagaagaaca gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc
     7501 cagatcctga aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg
     7561 tactacctgc agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg
     7621 tccgactacg atgtggacca tatcgtgcct cagagctttc tgaaggacga ctccatcgac
     7681 aacaaggtgc tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa
     7741 gaggtcgtga agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc
     7801 cagagaaagt tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag
     7861 gccggcttca tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag
     7921 atcctggact cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg
     7981 aaagtgatca ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac
     8041 aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg
     8101 ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac
     8161 aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc
     8221 gccaagtact tcttctacag caacatcatg aactttttca agaccgagat taccctggcc
     8281 aacggcgaga tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg
     8341 tgggataagg gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat
     8401 atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag
     8461 aggaacagcg ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc
     8521 ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag
     8581 tccaagaaac tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc
     8641 ttcgagaaga atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac
     8701 ctgatcatca agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg
     8761 ctggcctctg ccggcgaact gcagaaggga aacgaactgg ccctgccctc caaatatgtg
     8821 aacttcctgt acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag
     8881 cagaaacagc tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc
     8941 agcgagttct ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc
     9001 tacaacaagc accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt
     9061 accctgacca atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg
     9121 aagaggtaca ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc
     9181 ggcctgtacg agacacggat cgacctgtct cagctgggag gcgacaaaag gccggcggcc
     9241 acgaaaaagg ccggccaggc aaaaaagaaa aagtaactcg agggtggtgg tagcgattat
     9301 aaagatgatg atgataaagg cctgaacgat atctttgaag cccagaaaat tgaatggcac
     9361 gagtaaaagc ttgtcgagca ccaccaccac caccactgag atccggctgc taacaaagcc
     9421 cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg
     9481 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cggattggcg
     9541 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     9601 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     9661 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     9721 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     9781 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     9841 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     9901 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     9961 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
    10021 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
    10081 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
    10141 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
    10201 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
    10261 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
    10321 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
    10381 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
    10441 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
    10501 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
    10561 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
    10621 ggaaccggag ct
//
