LOCUS       sequence-360110-        9892 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FlpON knock-in for GluN1-smFP-V5 (amino acid position: A20).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9892)
  AUTHORS   Droogers WJ, Willems J, MacGillavry HD, de Jong APH
  TITLE     Duplex Labeling and Manipulation of Neuronal Proteins Using 
            Sequential CRISPR/Cas9 Gene Editing.
  JOURNAL   eNeuro. 2022 Jul 18;9(4):ENEURO.0056-22.2022. doi: 
            10.1523/ENEURO.0056-22.2022.
  PUBMED    35851300
REFERENCE   2  (bases 1 to 9892)
  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: 
            "10.1523/ENEURO.0056-22"; journalName: "eNeuro"; date: "2022-07-18- 
            18"; volume: "9"; issue: "4"; pages: "ENEURO.0056-22.2022"
COMMENT     Sequence Label: pOC4 Grin1-smFP-V5 KI
FEATURES             Location/Qualifiers
     source          1..9892
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        62..310
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     62..82
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     233..252
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     protein_bind    349..382
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    421..454
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     regulatory      649..658
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             658..699
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             709..750
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             760..801
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1333..1374
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1384..1425
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1444..1485
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1495..1536
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1750..1791
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1801..1842
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1852..1893
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     enhancer        1960..2245
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        2247..2524
                     /label=chicken beta-actin promoter
     intron          2525..2752
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     regulatory      2764..2773
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2773..2796
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2800..2820
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2830..6930
                     /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             2830..6930
                     /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             6934..6954
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6994..7011)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7000..7207
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   7216..7356
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   7216..7345
                     /label=AAV2 ITR
     rep_origin      7431..7886
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7518..7537)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7728..7749
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7903..7922)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8022..8044
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8082..8100)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8168..8272
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8273..8341,8342..9133)
                     /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(8491..8510)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9304..9892
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9793..9812
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 tgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg ctgttagaga
      121 gataattgga attaatttga ctgtaaacac aaagatatta gtacaaaata cgtgacgtag
      181 aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa tggactatca
      241 tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct tgtggaaagg
      301 acgaaacacc gcttggggtc gcaggcggcg cgtttaagag ctatgctgga agttcctatt
      361 ctctagaaag tataggaact tctttttttt ttgcggccgc ccagaattct ggcctgcagg
      421 gaagttccta ttctctagaa agtataggaa cttccagcat agcaagttta aataaggcta
      481 gtccgttatc aacttgaaaa agtggcaccg agtcggtgct tttttgtttt agagctagaa
      541 atagcaagtt aaaataaggc tagtccgttt ttagcgcgtg cgccaattct gcagacaaat
      601 ggctctagaa gcttcttggg gtcgcaggcg gcgctggcag ctagcggagc caccatggga
      661 aaacctatac cgaaccccct ccttggactt gatagtactc agcagcaggg taagcccatt
      721 ccaaacccac ttctcggtct ggatagcaca cagcaacaag gtaagcctat ccctaaccct
      781 ctgttgggcc tggactccac gggccaaggt gtgagtaagg gcgaagaact gtttaccggc
      841 gtggtcccaa ttctggttga gctcgacggt gacgttaacg gacacaagtt tagtgtgcgc
      901 ggagaaggag agggagatgc caccaacgga aaattgacgc tgaagtttat atgcaccacg
      961 ggaaagctgc cggtgccgtg gccgaccttg gttacaaccc tgggtggcgg agtccagtgt
     1021 ttcagccgtt acccagacca catgaagcag catgacttct ttaagagtgc tatgccggag
     1081 ggatacgtgc aggaacggac gatcagtttc aaagatgacg gtacttacaa gacgcgagcc
     1141 gaggtgaagt tcgagggaga caccttggtg aacaggatag aacttaaggg aattgatttc
     1201 aaggaagacg gtaatattct gggtcacaag ctggaataca actttaacag ccataacgtc
     1261 tacattactg ccgacaagca gaagaacggc ataaaggcga atttcaaaat tcggcacaac
     1321 gtggaaggtg gtggcaagcc gatcccgaat cccctgctgg gcttggatag cacgcagcaa
     1381 cagggaaagc ccatccctaa ccccttgctg ggactcgaca gcaccggagg ccagcaaggc
     1441 ggtggcaaac cgatacccaa tcccctgctc ggcctggata gcacccagca gcaaggcaag
     1501 ccaatcccca accctctgct gggcctcgac tcgacaggtg gcgatggaag tgttcaactg
     1561 gcggaccact atcagcagaa tacgccgatc ggagatggcc ccgtgctgtt gccggataat
     1621 cactatctgt cgactcagag cgttctgagt aaggacccaa acgaaaaacg agatcacatg
     1681 gtgttgttgg agttcgtgac cgccgccggc attacgttgg gaatggacga gctgtacaag
     1741 ggccaaggag gcaagcccat accaaatccc ttgctgggcc tcgacagcac tcagcaacaa
     1801 ggaaagccaa taccaaatcc actgttgggc ctcgacagta cgcaacagca gggcaagccc
     1861 atcccgaacc cgttgctggg actggatagt acctcaagcg cttccttggg gtcgcaggcg
     1921 gcgctggctc gaggatccta ggacgcgtcc agaggtaccc gttacataac ttacggtaaa
     1981 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaatag taacgccaat
     2041 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     2101 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     2161 cgcctggcat tgtgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     2221 cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct tcactctccc
     2281 catctccccc ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc
     2341 agcgatgggg gcgggggggg ggggggggcg cgcgccrggs ggggsggggs ggggsgrggg
     2401 gsggggsggg gsgaggcgga gaggkgcggc ggcagccaat cagagcggcg cgctccgaaa
     2461 gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg
     2521 ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg
     2581 cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc
     2641 tcctccgggc tgtaattagc tgagcaagag gtaagggttt aagggatggt tggttggtgg
     2701 ggtattaatg tttaattacc tggagcacct gcctgaaatc actttttttc aggttggacc
     2761 ggtgccacca tggattacaa agatgatgac gataaatcgc cgaagaaaaa gcgcaaggtc
     2821 gaagcgtccg acaagaagta cagcatcggc ctggacatcg gcaccaactc tgtgggctgg
     2881 gccgtgatca ccgacgagta caaggtgccc agcaagaaat tcaaggtgct gggcaacacc
     2941 gaccggcaca gcatcaagaa gaacctgatc ggagccctgc tgttcgacag cggcgaaaca
     3001 gccgaggcca cccggctgaa gagaaccgcc agaagaagat acaccagacg gaagaaccgg
     3061 atctgctatc tgcaagagat cttcagcaac gagatggcca aggtggacga cagcttcttc
     3121 cacagactgg aagagtcctt cctggtggaa gaggataaga agcacgagcg gcaccccatc
     3181 ttcggcaaca tcgtggacga ggtggcctac cacgagaagt accccaccat ctaccacctg
     3241 agaaagaaac tggtggacag caccgacaag gccgacctgc ggctgatcta tctggccctg
     3301 gcccacatga tcaagttccg gggccacttc ctgatcgagg gcgacctgaa ccccgacaac
     3361 agcgacgtgg acaagctgtt catccagctg gtgcagacct acaaccagct gttcgaggaa
     3421 aaccccatca acgccagcgg cgtggacgcc aaggccatcc tgtctgccag actgagcaag
     3481 agcagacggc tggaaaatct gatcgcccag ctgcccggcg agaagaagaa tggcctgttc
     3541 ggaaacctga ttgccctgag cctgggcctg acccccaact tcaagagcaa cttcgacctg
     3601 gccgaggatg ccaaactgca gctgagcaag gacacctacg acgacgacct ggacaacctg
     3661 ctggcccaga tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa cctgtccgac
     3721 gccatcctgc tgagcgacat cctgagagtg aacaccgaga tcaccaaggc ccccctgagc
     3781 gcctctatga tcaagagata cgacgagcac caccaggacc tgaccctgct gaaagctctc
     3841 gtgcggcagc agctgcctga gaagtacaaa gagattttct tcgaccagag caagaacggc
     3901 tacgccggct acattgacgg cggagccagc caggaagagt tctacaagtt catcaagccc
     3961 atcctggaaa agatggacgg caccgaggaa ctgctcgtga agctgaacag agaggacctg
     4021 ctgcggaagc agcggacctt cgacaacggc agcatccccc accagatcca cctgggagag
     4081 ctgcacgcca ttctgcggcg gcaggaagat ttttacccat tcctgaagga caaccgggaa
     4141 aagatcgaga agatcctgac cttccgcatc ccctactacg tgggccctct ggccagggga
     4201 aacagcagat tcgcctggat gaccagaaag agcgaggaaa ccatcacccc ctggaacttc
     4261 gaggaagtgg tggacaaggg cgcttccgcc cagagcttca tcgagcggat gaccaacttc
     4321 gataagaacc tgcccaacga gaaggtgctg cccaagcaca gcctgctgta cgagtacttc
     4381 accgtgtata acgagctgac caaagtgaaa tacgtgaccg agggaatgag aaagcccgcc
     4441 ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc tgttcaagac caaccggaaa
     4501 gtgaccgtga agcagctgaa agaggactac ttcaagaaaa tcgagtgctt cgactccgtg
     4561 gaaatctccg gcgtggaaga tcggttcaac gcctccctgg gcacatacca cgatctgctg
     4621 aaaattatca aggacaagga cttcctggac aatgaggaaa acgaggacat tctggaagat
     4681 atcgtgctga ccctgacact gtttgaggac agagagatga tcgaggaacg gctgaaaacc
     4741 tatgcccacc tgttcgacga caaagtgatg aagcagctga agcggcggag atacaccggc
     4801 tggggcaggc tgagccggaa gctgatcaac ggcatccggg acaagcagtc cggcaagaca
     4861 atcctggatt tcctgaagtc cgacggcttc gccaacagaa acttcatgca gctgatccac
     4921 gacgacagcc tgacctttaa agaggacatc cagaaagccc aggtgtccgg ccagggcgat
     4981 agcctgcacg agcacattgc caatctggcc ggcagccccg ccattaagaa gggcatcctg
     5041 cagacagtga aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa gcccgagaac
     5101 atcgtgatcg aaatggccag agagaaccag accacccaga agggacagaa gaacagccgc
     5161 gagagaatga agcggatcga agagggcatc aaagagctgg gcagccagat cctgaaagaa
     5221 caccccgtgg aaaacaccca gctgcagaac gagaagctgt acctgtacta cctgcagaat
     5281 gggcgggata tgtacgtgga ccaggaactg gacatcaacc ggctgtccga ctacgatgtg
     5341 gaccatatcg tgcctcagag ctttctgaag gacgactcca tcgacaacaa ggtgctgacc
     5401 agaagcgaca agaaccgggg caagagcgac aacgtgccct ccgaagaggt cgtgaagaag
     5461 atgaagaact actggcggca gctgctgaac gccaagctga ttacccagag aaagttcgac
     5521 aatctgacca aggccgagag aggcggcctg agcgaactgg ataaggccgg cttcatcaag
     5581 agacagctgg tggaaacccg gcagatcaca aagcacgtgg cacagatcct ggactcccgg
     5641 atgaacacta agtacgacga gaatgacaag ctgatccggg aagtgaaagt gatcaccctg
     5701 aagtccaagc tggtgtccga tttccggaag gatttccagt tttacaaagt gcgcgagatc
     5761 aacaactacc accacgccca cgacgcctac ctgaacgccg tcgtgggaac cgccctgatc
     5821 aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg actacaaggt gtacgacgtg
     5881 cggaagatga tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa gtacttcttc
     5941 tacagcaaca tcatgaactt tttcaagacc gagattaccc tggccaacgg cgagatccgg
     6001 aagcggcctc tgatcgagac aaacggcgaa accggggaga tcgtgtggga taagggccgg
     6061 gattttgcca ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt gaaaaagacc
     6121 gaggtgcaga caggcggctt cagcaaagag tctatcctgc ccaagaggaa cagcgataag
     6181 ctgatcgcca gaaagaagga ctgggaccct aagaagtacg gcggcttcga cagccccacc
     6241 gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa gaaactgaag
     6301 agtgtgaaag agctgctggg gatcaccatc atggaaagaa gcagcttcga gaagaatccc
     6361 atcgactttc tggaagccaa gggctacaaa gaagtgaaaa aggacctgat catcaagctg
     6421 cctaagtact ccctgttcga gctggaaaac ggccggaaga gaatgctggc ctctgccggc
     6481 gaactgcaga agggaaacga actggccctg ccctccaaat atgtgaactt cctgtacctg
     6541 gccagccact atgagaagct gaagggctcc cccgaggata atgagcagaa acagctgttt
     6601 gtggaacagc acaagcacta cctggacgag atcatcgagc agatcagcga gttctccaag
     6661 agagtgatcc tggccgacgc taatctggac aaagtgctgt ccgcctacaa caagcaccgg
     6721 gataagccca tcagagagca ggccgagaat atcatccacc tgtttaccct gaccaatctg
     6781 ggagcccctg ccgccttcaa gtactttgac accaccatcg accggaagag gtacaccagc
     6841 accaaagagg tgctggacgc caccctgatc caccagagca tcaccggcct gtacgagaca
     6901 cggatcgacc tgtctcagct gggaggcgac agccccaaga agaagagaaa ggtggaggcc
     6961 agctaagaat tctaactaga gctcgctgat cagcctcgac tgtgccttct agttgccagc
     7021 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     7081 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     7141 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagagaat agcaggcatg
     7201 ctggggagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     7261 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     7321 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
     7381 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     7441 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     7501 cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     7561 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     7621 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     7681 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     7741 ccaaactgga acaacactca actctatctc gggctattct tttgatttat aagggatttt
     7801 gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     7861 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     7921 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     7981 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     8041 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     8101 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     8161 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     8221 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     8281 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     8341 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     8401 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     8461 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     8521 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     8581 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     8641 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     8701 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     8761 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     8821 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     8881 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     8941 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtggaa gccgcggtat
     9001 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     9061 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     9121 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     9181 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     9241 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     9301 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     9361 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     9421 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     9481 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     9541 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     9601 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     9661 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     9721 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     9781 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     9841 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aa
//
