LOCUS       sequence-244715-        9501 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses SpCas9 fused to DHFR domains on both the N-terminus and an
            internal loop in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9501)
  AUTHORS   Manna D, Maji B, Gangopadhyay SA, Cox KJ, Zhou Q, Law BK, Mazitschek
            R, Choudhary A
  TITLE     A Singular System with Precise Dosing and Spatiotemporal Control of 
            CRISPR-Cas9.
  JOURNAL   Angew Chem Int Ed Engl. 2019 May 6;58(19):6285-6289. doi: 
            10.1002/anie.201900788. Epub 2019 Apr 2.
  PUBMED    30834641
REFERENCE   2  (bases 1 to 9501)
  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: 
            "10.1002/anie.201900788"; journalName: "Angew Chem Int Ed Engl"; 
            date: "2019-05-6- 6"; volume: "58"; issue: "19"; pages: "6285-6289"
COMMENT     Sequence Label: pX330-NL-DHFR-SpCas9
FEATURES             Location/Qualifiers
     source          1..9501
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        268..343
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        440..725
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          1005..1232
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1244..1253
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1253..1318
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1325..1345
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1376..1849
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="ISLIAALAVDYVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRHT
                     WESIGRPLPGRKNIILSSQPSTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVIEQFLP
                     KAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     CDS             3968..6466
                     /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             6467..6514
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(6542..6559)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6548..6755
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6764..6904
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6764..6893
                     /label=AAV2 ITR
     rep_origin      6979..7434
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7066..7085)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7276..7297
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7451..7470)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7570..7592
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7630..7648)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7716..7820
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7821..7889,7890..8681)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(8039..8058)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8852..9440
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9341..9360
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg ggtcttcgag aagacctgtt ttagagctag aaatagcaag ttaaaataag
      301 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg ttttagagct
      361 agaaatagca agttaaaata aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac
      421 aaatggctct agaggtaccc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      481 ccaacgaccc ccgcccattg acgtcaatag taacgccaat agggactttc cattgacgtc
      541 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      601 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tgtgcccagt
      661 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      721 ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac
      781 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgaggggcg
      841 gggcggggcg aggcggrgag gkgcggcggs rgccaaknsr gmggggcgsg scgmgrsggw
      901 smkktkykgs ggcrgccaat cagrgcggcg cgctccgaaa gtttcctttt atggcgaggc
      961 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct
     1021 gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga
     1081 ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc
     1141 tgagcaagag gtaagggttt aagggatggt tggttggtgg ggtattaatg tttaattacc
     1201 tggagcacct gcctgaaatc actttttttc aggttggacc ggtgccacca tggactataa
     1261 ggaccacgac ggagactaca aggatcatga tattgattac aaagacgatg acgataagat
     1321 ggccccaaag aagaagcgga aggtcggtat ccacggagtc ccagcagccg gatccatcag
     1381 tctgattgcg gcgttagcgg tagattacgt tatcggcatg gaaaacgcca tgccgtggaa
     1441 cctgcctgcc gatctcgcct ggtttaaacg caacacctta aataaacccg tgattatggg
     1501 ccgccatacc tgggaatcaa tcggtcgtcc gttgccagga cgcaaaaata ttatcctcag
     1561 cagtcaaccg agtacggacg atcgcgtaac gtgggtgaag tcggtggatg aagccatcgc
     1621 ggcgtgtggt gacgtaccag aaatcatggt gattggcggc ggtcgcgtta ttgaacagtt
     1681 cttgccaaaa gcgcaaaaac tgtatctgac gcatatcgac gcagaagtgg aaggcgacac
     1741 ccatttcccg gattacgagc cggatgactg ggaatcggta ttcagcgaat tccacgatgc
     1801 tgatgcgcag aactctcaca gctattgctt tgagattctg gagcggcgag gaagcggaag
     1861 cggatccgac aagaagtaca gcatcggcct ggacatcggc accaactctg tgggctgggc
     1921 cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga
     1981 ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc
     2041 cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat
     2101 ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca
     2161 cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt
     2221 cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag
     2281 aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc
     2341 ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag
     2401 cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa
     2461 ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag
     2521 cagacggctg gaaaatctga tcgcccagct gcccggcgga agtggcagca tcagcctgat
     2581 cgctgctctg gcagtagact acgtgatagg aatggagaat gcaatgccct ggaatctgcc
     2641 cgcagacctg gcttggttta aaagaaacac gcttaataaa ccagtaatca tgggaagaca
     2701 tacttgggag agtatagggc ggcccctgcc cggccggaaa aatattatac tctcaagtca
     2761 accgagcact gacgatagag tcacctgggt gaagtccgtt gatgaggcta tagccgcatg
     2821 tggagatgtg cctgaaatca tggtaatcgg tggtggaagg gtcatagagc aattcctccc
     2881 gaaggctcag aaactttatc ttacacatat agacgccgag gtggagggcg acacacattt
     2941 ccctgactat gaacccgatg attgggaaag cgtatttagt gaatttcacg acgccgacgc
     3001 acagaactct cacagttatt gcttcgagat tctggagcga cgaggcagtg gaagcgagaa
     3061 gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc ccaacttcaa
     3121 gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga
     3181 cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc
     3241 caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac
     3301 caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac
     3361 cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga
     3421 ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg aagagttcta
     3481 caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct
     3541 gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca
     3601 gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct
     3661 gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg
     3721 ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat
     3781 caccccctgg aacttcgagg aagtggtgga caagggcgct tccgcccaga gcttcatcga
     3841 gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct
     3901 gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg
     3961 aatgagaaag cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt
     4021 caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga
     4081 gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac
     4141 ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga
     4201 ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga
     4261 ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg
     4321 gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa
     4381 gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt
     4441 catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt
     4501 gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat
     4561 taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg
     4621 gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg
     4681 acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag
     4741 ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct
     4801 gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct
     4861 gtccgactac gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg actccatcga
     4921 caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg tgccctccga
     4981 agaggtcgtg aagaagatga agaactactg gcggcagctg ctgaacgcca agctgattac
     5041 ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa
     5101 ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca
     5161 gatcctggac tcccggatga acactaagta cgacgagaat gacaagctga tccgggaagt
     5221 gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta
     5281 caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga acgccgtcgt
     5341 gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt acggcgacta
     5401 caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg gcaaggctac
     5461 cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc
     5521 caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt
     5581 gtgggataag ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa
     5641 tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa
     5701 gaggaacagc gataagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg
     5761 cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa
     5821 gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag
     5881 cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga
     5941 cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat
     6001 gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt
     6061 gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga
     6121 gcagaaacag ctgtttgtgg aacagcacaa gcactacctg gacgagatca tcgagcagat
     6181 cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaaag tgctgtccgc
     6241 ctacaacaag caccgggata agcccatcag agagcaggcc gagaatatca tccacctgtt
     6301 taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg
     6361 gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac
     6421 cggcctgtac gagacacgga tcgacctgtc tcagctggga ggcgacaaaa ggccggcggc
     6481 cacgaaaaag gccggccagg caaaaaagaa aaagtaagaa ttcctagagc tcgctgatca
     6541 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     6601 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     6661 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     6721 gaggattggg aagagaatag caggcatgct ggggagcggc cgcaggaacc cctagtgatg
     6781 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     6841 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
     6901 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     6961 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     7021 ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct
     7081 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     7141 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     7201 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     7261 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg
     7321 gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc
     7381 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     7441 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     7501 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     7561 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     7621 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     7681 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     7741 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     7801 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     7861 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     7921 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     7981 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     8041 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     8101 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     8161 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     8221 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     8281 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     8341 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     8401 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     8461 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     8521 gagccggtga gcgtggaagc cgcggtatca ttgcagcact ggggccagat ggtaagccct
     8581 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     8641 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     8701 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     8761 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     8821 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     8881 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     8941 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     9001 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     9061 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     9121 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     9181 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     9241 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     9301 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     9361 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     9421 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     9481 gctggccttt tgctcacatg t
//
