LOCUS       sequence-422954-       10505 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To express a single-guide RNA under a U6 promoter, accompanied by 
            the expression of eCas9 and the affinity sorting tag TST-EGFP-GPIDAF
            under separate promoters..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10505)
  AUTHORS   Yang L, Cui L, Ma S, Zuo Q, Huang Q
  TITLE     A Gene Transfer-Positive Cell Sorting System Utilizing 
            Membrane-Anchoring Affinity Tag.
  JOURNAL   Front Bioeng Biotechnol. 2022 Jun 16;10:930966. doi: 
            10.3389/fbioe.2022.930966. eCollection 2022.
  PUBMED    35782508
REFERENCE   2  (bases 1 to 10505)
  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: "Front 
            Bioeng Biotechnol."; date: "2022-06-16"; volume: "10:930966. doi"; 
            pages: " 10.3389/fbioe.2022.930966. eCollection 2022"
COMMENT     Sequence Label: pCRISPR/eCas9-GPIDAF
FEATURES             Location/Qualifiers
     source          1..10505
                     /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"
     promoter        727..1001
                     /label=chicken beta-actin promoter
     intron          1002..1229
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      1241..1250
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1250..1315
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             1322..1342
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1367..5467
                     /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
                     KTYAHLFDDKVMKQLKRRRYTGWGALSRKLINGIRDKQSGKTILDFLKSDGFANRNFMA
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRAITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5468..5515
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     primer_bind     complement(5543..5560)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5549..5756
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        5820..6123
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6124..6327
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6278..6298
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(6470..6475,6476..6499,6500..6535,6536..6559)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     regulatory      6566..6575
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6572..6574,6575..6577,6578..7288)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(6572..6574,6575..6577,6578..7288)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6878..6897)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     7225..7246
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    7454..7510
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    7634..7755
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7671..7690)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7725..7744
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   7768..7908
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7768..7897
                     /label=AAV2 ITR
     rep_origin      7983..8438
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8070..8089)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8280..8301
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(8455..8474)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8574..8596
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8634..8652)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8720..8824
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8825..8893,8894..9685)
                     /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(9043..9062)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9856..10444
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10345..10364
                     /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 gggggggggg
      841 gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg gggcggggcg aggcggagag
      901 gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc
      961 ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc
     1021 ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg
     1081 cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagctga
     1141 gcaagaggta agggtttaag ggatggttgg ttggtggggt attaatgttt aattacctgg
     1201 agcacctgcc tgaaatcact ttttttcagg ttggaccggt gccaccatgg actataagga
     1261 ccacgacgga gactacaagg atcatgatat tgattacaaa gacgatgacg ataagatggc
     1321 cccaaagaag aagcggaagg tcggtatcca cggagtccca gcagccgaca agaagtacag
     1381 catcggcctg gacatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa
     1441 ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa
     1501 cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag
     1561 aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt
     1621 cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag agtccttcct
     1681 ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt
     1741 ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac
     1801 cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg
     1861 ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat
     1921 ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt
     1981 ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat
     2041 cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg ccctgagcct
     2101 gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct
     2161 gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta
     2221 cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct
     2281 gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga
     2341 cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa
     2401 gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca ttgacggcgg
     2461 agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac
     2521 cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga
     2581 caacggcagc atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca
     2641 ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt
     2701 ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac
     2761 cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc
     2821 ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa
     2881 ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg agctgaccaa
     2941 agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa
     3001 ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga
     3061 ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg
     3121 gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt
     3181 cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt
     3241 tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa
     3301 agtgatgaag cagctgaagc ggcggagata caccggctgg ggcgcgctga gccggaagct
     3361 gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga
     3421 cggcttcgcc aacagaaact tcatggcgct gatccacgac gacagcctga cctttaaaga
     3481 ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa
     3541 tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga
     3601 gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga
     3661 gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga
     3721 gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct
     3781 gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca
     3841 ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc ctcagagctt
     3901 tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga accggggcaa
     3961 gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcggcagct
     4021 gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg ccgagagagg
     4081 cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg aaacccgggc
     4141 gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa
     4201 tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt
     4261 ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga
     4321 cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag
     4381 cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga
     4441 gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt
     4501 caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa
     4561 cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg tgcggaaagt
     4621 gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag
     4681 caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa agaaggactg
     4741 ggaccctaag aagtacggcg gcttcgatag ccccaccgtg gcctattctg tgctggtggt
     4801 ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc tgctggggat
     4861 caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg
     4921 ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct
     4981 ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact
     5041 ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa
     5101 gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca agcactacct
     5161 ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa
     5221 tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca gagagcaggc
     5281 cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta
     5341 ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc tggacgccac
     5401 cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg
     5461 aggcgacaaa aggccggcgg ccacgaaaaa ggccggccag gcaaaaaaga aaaagtaaga
     5521 attcctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     5581 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     5641 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     5701 gtggggcagg acagcaaggg ggaggattgg gaagagaata gcaggcatgc tggggagcgg
     5761 ccttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     5821 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     5881 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     5941 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     6001 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     6061 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     6121 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     6181 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     6241 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     6301 cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg ctagcgctac
     6361 cggtaccatg accgtggcta gacccagcgt gcctgctgcc ctgcctctgc tgggagagct
     6421 gcctagactg ctgctcctgg tgctcctgtg cctgcctgct gtgtggggaa gcgcctggag
     6481 ccaccctcag ttcgagaaag gaggaggcag cggcggcgga agcggaggct ccgcttggtc
     6541 ccacccacag ttcgaaaagc cggtcgccac catggtgagc aagggcgagg agctgttcac
     6601 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     6661 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     6721 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     6781 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     6841 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     6901 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     6961 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     7021 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     7081 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     7141 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     7201 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     7261 cactctcggc atggacgagc tgtacaagtc cggccggact cagatcagcc ggaccaccaa
     7321 gcacttccac gagaccaccc ctaacaaggg cagcggcacc accagcggca ccacccggct
     7381 gctgagcggc cacacctgct tcaccctgac cggcctgctg ggcaccctgg tgaccatggg
     7441 cctgctgacc tagagatctc gagctcaagc ttcgaattct gcagtcgacg gtaccgcggg
     7501 cccgggatcc accggatcta gataactgat cataatcagc cataccacat ttgtagaggt
     7561 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     7621 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     7681 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     7741 catcaatgta tcttaacgcg tggccgcagg aacccctagt gatggagttg gccactccct
     7801 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     7861 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     7921 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     7981 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     8041 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     8101 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     8161 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     8221 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     8281 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     8341 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     8401 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     8461 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     8521 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     8581 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     8641 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     8701 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     8761 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     8821 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     8881 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     8941 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     9001 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     9061 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     9121 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     9181 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     9241 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     9301 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     9361 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     9421 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     9481 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     9541 aagccgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     9601 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     9661 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     9721 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     9781 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     9841 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     9901 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     9961 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
    10021 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
    10081 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
    10141 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
    10201 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
    10261 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
    10321 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
    10381 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
    10441 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
    10501 catgt
//
