LOCUS       sequence-350715-        9451 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pSpCas9(BB)-2A-tCD19 (pDU1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9451)
  AUTHORS   Ramachandran H, Martins S, Kontarakis Z, Krutmann J, Rossi A
  TITLE     Fast but not furious: a streamlined selection method for 
            genome-edited cells.
  JOURNAL   Life Sci Alliance. 2021 Apr 26;4(6). pii: 4/6/e202101051. doi: 
            10.26508/lsa.202101051. Print 2021 Jun.
  PUBMED    33903218
REFERENCE   2  (bases 1 to 9451)
  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: "Life Sci 
            Alliance."; date: "2021-04-26"; volume: "4(6). pii: 4/6/e202101051. 
            doi"; pages: " 10.26508/lsa.202101051. Print 2021 Jun"
COMMENT     Sequence Label: pDU1
FEATURES             Location/Qualifiers
     source          1..9451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        82..330
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     82..102
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     253..272
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        349..424
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        521..806
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     intron          970..1197
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1209..1218
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1218..1283
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1290..1310
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1335..5435
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             1335..5435
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5436..5483
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             5490..5543
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     primer_bind     complement(6573..6590)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6579..6786
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   6795..6935
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6795..6924
                     /label=AAV2 ITR
     rep_origin      7010..7465
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7097..7116)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7307..7328
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7482..7501)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7601..7623
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7661..7679)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7747..7851
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7852..7920,7921..8712)
                     /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(8070..8089)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8883..9451,1..20)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9372..9391
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
       61 gctggccttt tgctcacatg tgagggccta tttcccatga ttccttcata tttgcatata
      121 cgatacaagg ctgttagaga gataattgga attaatttga ctgtaaacac aaagatatta
      181 gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta
      241 tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct
      301 ttatatatct tgtggaaagg acgaaacacc gggtcttcga gaagacctgt tttagagcta
      361 gaaatagcaa gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
      421 gtgctttttt gttttagagc tagaaatagc aagttaaaat aaggctagtc cgtttttagc
      481 gcgtgcgcca attctgcaga caaatggctc tagaggtacc cgttacataa cttacggtaa
      541 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata gtaacgccaa
      601 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      661 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      721 ccgcctggca ttgtgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      781 acgtattagt catcgctatt accatggtcg aggtgagccc cacgttctgc ttcactctcc
      841 ccatctcccc cccctcccag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc
      901 cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg
      961 cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg
     1021 cgccgcccgc cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc
     1081 ccttctcctc cgggctgtaa ttagctgagc aagaggtaag ggtttaaggg atggttggtt
     1141 ggtggggtat taatgtttaa ttacctggag cacctgcctg aaatcacttt ttttcaggtt
     1201 ggaccggtgc caccatggac tataaggacc acgacggaga ctacaaggat catgatattg
     1261 attacaaaga cgatgacgat aagatggccc caaagaagaa gcggaaggtc ggtatccacg
     1321 gagtcccagc agccgacaag aagtacagca tcggcctgga catcggcacc aactctgtgg
     1381 gctgggccgt gatcaccgac gagtacaagg tgcccagcaa gaaattcaag gtgctgggca
     1441 acaccgaccg gcacagcatc aagaagaacc tgatcggagc cctgctgttc gacagcggcg
     1501 aaacagccga ggccacccgg ctgaagagaa ccgccagaag aagatacacc agacggaaga
     1561 accggatctg ctatctgcaa gagatcttca gcaacgagat ggccaaggtg gacgacagct
     1621 tcttccacag actggaagag tccttcctgg tggaagagga taagaagcac gagcggcacc
     1681 ccatcttcgg caacatcgtg gacgaggtgg cctaccacga gaagtacccc accatctacc
     1741 acctgagaaa gaaactggtg gacagcaccg acaaggccga cctgcggctg atctatctgg
     1801 ccctggccca catgatcaag ttccggggcc acttcctgat cgagggcgac ctgaaccccg
     1861 acaacagcga cgtggacaag ctgttcatcc agctggtgca gacctacaac cagctgttcg
     1921 aggaaaaccc catcaacgcc agcggcgtgg acgccaaggc catcctgtct gccagactga
     1981 gcaagagcag acggctggaa aatctgatcg cccagctgcc cggcgagaag aagaatggcc
     2041 tgttcggaaa cctgattgcc ctgagcctgg gcctgacccc caacttcaag agcaacttcg
     2101 acctggccga ggatgccaaa ctgcagctga gcaaggacac ctacgacgac gacctggaca
     2161 acctgctggc ccagatcggc gaccagtacg ccgacctgtt tctggccgcc aagaacctgt
     2221 ccgacgccat cctgctgagc gacatcctga gagtgaacac cgagatcacc aaggcccccc
     2281 tgagcgcctc tatgatcaag agatacgacg agcaccacca ggacctgacc ctgctgaaag
     2341 ctctcgtgcg gcagcagctg cctgagaagt acaaagagat tttcttcgac cagagcaaga
     2401 acggctacgc cggctacatt gacggcggag ccagccagga agagttctac aagttcatca
     2461 agcccatcct ggaaaagatg gacggcaccg aggaactgct cgtgaagctg aacagagagg
     2521 acctgctgcg gaagcagcgg accttcgaca acggcagcat cccccaccag atccacctgg
     2581 gagagctgca cgccattctg cggcggcagg aagattttta cccattcctg aaggacaacc
     2641 gggaaaagat cgagaagatc ctgaccttcc gcatccccta ctacgtgggc cctctggcca
     2701 ggggaaacag cagattcgcc tggatgacca gaaagagcga ggaaaccatc accccctgga
     2761 acttcgagga agtggtggac aagggcgctt ccgcccagag cttcatcgag cggatgacca
     2821 acttcgataa gaacctgccc aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt
     2881 acttcaccgt gtataacgag ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc
     2941 ccgccttcct gagcggcgag cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc
     3001 ggaaagtgac cgtgaagcag ctgaaagagg actacttcaa gaaaatcgag tgcttcgact
     3061 ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca taccacgatc
     3121 tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag gacattctgg
     3181 aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag gaacggctga
     3241 aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg cggagataca
     3301 ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag cagtccggca
     3361 agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc atgcagctga
     3421 tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg tccggccagg
     3481 gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt aagaagggca
     3541 tcctgcagac agtgaaggtg gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg
     3601 agaacatcgt gatcgaaatg gccagagaga accagaccac ccagaaggga cagaagaaca
     3661 gccgcgagag aatgaagcgg atcgaagagg gcatcaaaga gctgggcagc cagatcctga
     3721 aagaacaccc cgtggaaaac acccagctgc agaacgagaa gctgtacctg tactacctgc
     3781 agaatgggcg ggatatgtac gtggaccagg aactggacat caaccggctg tccgactacg
     3841 atgtggacca tatcgtgcct cagagctttc tgaaggacga ctccatcgac aacaaggtgc
     3901 tgaccagaag cgacaagaac cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga
     3961 agaagatgaa gaactactgg cggcagctgc tgaacgccaa gctgattacc cagagaaagt
     4021 tcgacaatct gaccaaggcc gagagaggcg gcctgagcga actggataag gccggcttca
     4081 tcaagagaca gctggtggaa acccggcaga tcacaaagca cgtggcacag atcctggact
     4141 cccggatgaa cactaagtac gacgagaatg acaagctgat ccgggaagtg aaagtgatca
     4201 ccctgaagtc caagctggtg tccgatttcc ggaaggattt ccagttttac aaagtgcgcg
     4261 agatcaacaa ctaccaccac gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc
     4321 tgatcaaaaa gtaccctaag ctggaaagcg agttcgtgta cggcgactac aaggtgtacg
     4381 acgtgcggaa gatgatcgcc aagagcgagc aggaaatcgg caaggctacc gccaagtact
     4441 tcttctacag caacatcatg aactttttca agaccgagat taccctggcc aacggcgaga
     4501 tccggaagcg gcctctgatc gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg
     4561 gccgggattt tgccaccgtg cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa
     4621 agaccgaggt gcagacaggc ggcttcagca aagagtctat cctgcccaag aggaacagcg
     4681 ataagctgat cgccagaaag aaggactggg accctaagaa gtacggcggc ttcgacagcc
     4741 ccaccgtggc ctattctgtg ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac
     4801 tgaagagtgt gaaagagctg ctggggatca ccatcatgga aagaagcagc ttcgagaaga
     4861 atcccatcga ctttctggaa gccaagggct acaaagaagt gaaaaaggac ctgatcatca
     4921 agctgcctaa gtactccctg ttcgagctgg aaaacggccg gaagagaatg ctggcctctg
     4981 ccggcgaact gcagaaggga aacgaactgg ccctgccctc caaatatgtg aacttcctgt
     5041 acctggccag ccactatgag aagctgaagg gctcccccga ggataatgag cagaaacagc
     5101 tgtttgtgga acagcacaag cactacctgg acgagatcat cgagcagatc agcgagttct
     5161 ccaagagagt gatcctggcc gacgctaatc tggacaaagt gctgtccgcc tacaacaagc
     5221 accgggataa gcccatcaga gagcaggccg agaatatcat ccacctgttt accctgacca
     5281 atctgggagc ccctgccgcc ttcaagtact ttgacaccac catcgaccgg aagaggtaca
     5341 ccagcaccaa agaggtgctg gacgccaccc tgatccacca gagcatcacc ggcctgtacg
     5401 agacacggat cgacctgtct cagctgggag gcgacaaaag gccggcggcc acgaaaaagg
     5461 ccggccaggc aaaaaagaaa aaggaattcg agggcagggg cagcctgctg acctgcggcg
     5521 acgtggagga gaaccccggc cccatgcccc cccccaggct gctgttcttc ctgctgttcc
     5581 tgacccccat ggaggtgagg cccgaggagc ccctggtggt gaaggtggag gagggcgaca
     5641 acgccgtgct gcagtgcctg aagggcacca gcgacggccc cacccagcag ctgacctgga
     5701 gcagggagag ccccctgaag cccttcctga agctgagcct gggcctgccc ggcctgggca
     5761 tccacatgag gcccctggcc atctggctgt tcatcttcaa cgtgagccag cagatgggcg
     5821 gcttctacct gtgccagccc ggccccccca gcgagaaggc ctggcagccc ggctggaccg
     5881 tgaacgtgga gggcagcggc gagctgttca ggtggaacgt gagcgacctg ggcggcctgg
     5941 gctgcggcct gaagaacagg agcagcgagg gccccagcag ccccagcggc aagctgatga
     6001 gccccaagct gtacgtgtgg gccaaggaca ggcccgagat ctgggagggc gagcccccct
     6061 gcctgccccc cagggacagc ctgaaccaga gcctgagcca ggacctgacc atggcccccg
     6121 gcagcaccct gtggctgagc tgcggcgtgc cccccgacag cgtgagcagg ggccccctga
     6181 gctggaccca cgtgcacccc aagggcccca agagcctgct gagcctggag ctgaaggacg
     6241 acaggcccgc cagggacatg tgggtgatgg agaccggcct gctgctgccc agggccaccg
     6301 cccaggacgc cggcaagtac tactgccaca ggggcaacct gaccatgagc ttccacctgg
     6361 agatcaccgc caggcccgtg ctgtggcact ggctgctgag gaccggcggc tggaaggtga
     6421 gcgccgtgac cctggcctac ctgatcttct gcctgtgcag cctggtgggc atcctgcacc
     6481 tgcagagggc cctggtgctg aggaggaaga ggaagaggat gaccgacccc accaggaggt
     6541 tctgagaatt ctaactagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
     6601 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     6661 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     6721 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagagaata gcaggcatgc
     6781 tggggagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     6841 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     6901 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct
     6961 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt
     7021 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     7081 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     7141 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     7201 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga
     7261 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     7321 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg
     7381 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     7441 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc
     7501 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     7561 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     7621 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt
     7681 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     7741 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     7801 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     7861 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     7921 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     7981 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     8041 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     8101 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     8161 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     8221 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     8281 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     8341 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     8401 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     8461 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     8521 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtggaag ccgcggtatc
     8581 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     8641 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     8701 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     8761 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     8821 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     8881 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     8941 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     9001 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     9061 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     9121 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     9181 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     9241 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     9301 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     9361 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     9421 cttgagcgtc gatttttgtg atgctcgtca g
//
