LOCUS       sequence-257280-       11472 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Endogenous tagging of beta-3 Tubulin: C-terminal (amino acid 
            position: STOP codon).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11472)
  AUTHORS   Willems J, de Jong APH, Scheefhals N, Mertens E, Catsburg LAE, 
            Poorthuis RB, de Winter F, Verhaagen J, Meye FJ, MacGillavry HD
  TITLE     ORANGE: A CRISPR/Cas9-based genome editing toolbox for epitope 
            tagging of endogenous proteins in neurons.
  JOURNAL   PLoS Biol. 2020 Apr 10;18(4):e3000665. doi: 
            10.1371/journal.pbio.3000665.
  PUBMED    32275651
REFERENCE   2  (bases 1 to 11472)
  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: "PLoS 
            Biol."; date: "2020-04-10"; volume: "18(4):e3000665. doi"; pages: " 
            10.1371/journal.pbio.3000665"
COMMENT     Sequence Label: pFUGW ORANGE Tubb3-GFP KI _ mCherry-KASH
FEATURES             Location/Qualifiers
     source          1..11472
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    277..394
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        436..684
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     436..456
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     607..626
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        704..779
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     CDS             join(906..908,909..1622)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(906..908,909..1619)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(948..969)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1209..1228)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1556..1577
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        1741..2952
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     primer_bind     2789..2808
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             2999..3709
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MTVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYE
                     GTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMN
                     FEDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGAL
                     KGEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYER
                     AEGRHSTGGMDELYK"
     primer_bind     complement(3147..3165)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3415..3434
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    4001..4589
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4054..4074)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4472..4483)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4592..4608)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4593..4609)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             5114..5294
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(5320..5337)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5326..5550
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5596..6024
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5683..5702)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5893..5914
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6033..6053)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6038..6367
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6218..6353
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6280..6299
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6415..6462
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6481..6855
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    6985..7106
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7022..7041)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7076..7095
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7155..7171)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7155..7171)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7168..7190)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7179..7195
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(7203..7209,7210..7227,7228..7233))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7248..7269
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7386..7403)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7557..8145)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7637..7656)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8316..9107,9108..9176))
                     /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     8939..8958
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9177..9281)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9356..9375)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        9547..9926
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9928..10126
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     10081..10101
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             10144..10324
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    10371..10496
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    10989..11222
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             11407..11451
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
ORIGIN
        1 aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt aggtttaaga
       61 atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc accattatcg
      121 tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat agaagaagaa
      181 ggtggagaga gagacagaga cagatccatt cgattagtga acggatcggc actgcgtgcg
      241 ccaattctgc agacaaatgg cagtattcat ccacaatttt aaaagaaaag gggggattgg
      301 ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga
      361 attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg acagcagaga
      421 tccagtttgg ttagtgaggg cctatttccc atgattcctt catatttgca tatacgatac
      481 aaggctgtta gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa
      541 aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt
      601 aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat
      661 atcttgtgga aaggacgaaa caccgctgcg agcaacttca cttgttttag agctagaaat
      721 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
      781 tttttgtttt agagctagaa atagcaagtt aaaataaggc tagtccgttt ttagcgcgtg
      841 cgccaattct gcagacaaat ggctctagaa gcttgctgcg agcaacttca cttgggggat
      901 caggcgtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
      961 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     1021 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     1081 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     1141 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     1201 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     1261 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     1321 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     1381 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     1441 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     1501 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     1561 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     1621 aatgtgctgc gagcaacttc acttgggctc gaggatccta ggacgcgtcc taggacgcgt
     1681 ccagaggtac ccgttacata acttacggta aatggcctaa cccgtgtcgg ctccagatct
     1741 ggcctccgcg ccgggttttg gcgcctcccg cgggcgcccc cctcctcacg gcgagcgctg
     1801 ccacgtcaga cgaagggcgc agcgagcgtc ctgatccttc cgcccggacg ctcaggacag
     1861 cggcccgctg ctcataagac tcggccttag aaccccagta tcagcagaag gacattttag
     1921 gacgggactt gggtgactct agggcactgg ttttctttcc agagagcgga acaggcgagg
     1981 aaaagtagtc ccttctcggc gattctgcgg agggatctcc gtggggcggt gaacgccgat
     2041 gattatataa ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg ggatttgggt
     2101 cgcggttctt gtttgtggat cgctgtgatc gtcacttggt gagtagcggg ctgctgggct
     2161 ggccggggct ttcgtggccg ccgggccgct cggtgggacg gaagcgtgtg gagagaccgc
     2221 caagggctgt agtctgggtc cgcgagcaag gttgccctga actgggggtt ggggggagcg
     2281 cagcaaaatg gcggctgttc ccgagtcttg aatggaagac gcttgtgagg cgggctgtga
     2341 ggtcgttgaa acaaggtggg gggcatggtg ggcggcaaga acccaaggtc ttgaggcctt
     2401 cgctaatgcg ggaaagctct tattcgggtg agatgggctg gggcaccatc tggggaccct
     2461 gacgtgaagt ttgtcactga ctggagaact cggtttgtcg tctgttgcgg gggcggcagt
     2521 tatggcggtg ccgttgggca gtgcacccgt acctttggga gcgcgcgccc tcgtcgtgtc
     2581 gtgacgtcac ccgttctgtt ggcttataat gcagggtggg gccacctgcc ggtaggtgtg
     2641 cggtaggctt ttctccgtcg caggacgcag ggttcgggcc tagggtaggc tctcctgaat
     2701 cgacaggcgc cggacctctg gtgaggggag ggataagtga ggcgtcagtt tctttggtcg
     2761 gttttatgta cctatcttct taagtagctg aagctccggt tttgaactat gcgctcgggg
     2821 ttggcgagtg tgttttgtga agttttttag gcaccttttg aaatgtaatc atttgggtca
     2881 atatgtaatt ttcagtgtta gactagtaaa ttgtccgcta aattctggcc gtttttggct
     2941 tttttgttag acgaagcttg ggctgcaggt cgactctaga ggatccccgg gtaccggtat
     3001 gacggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
     3061 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     3121 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
     3181 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
     3241 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
     3301 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
     3361 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     3421 aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
     3481 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     3541 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     3601 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     3661 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt ccggactcag
     3721 atctcgagag gaggaggagg agacagacag caggatgccc cacctcgaca gccccggcag
     3781 ctcccagccg agacgctcct tcctctcaag ggtgatcagg gcagcgctac cgttgcagct
     3841 gcttctgctg ctgctgctgc tcctggcctg cctgctacct gcctctgaag atgactacag
     3901 ctgcacccag gccaacaact ttgcccgatc cttctacccc atgctgcggt acaccaacgg
     3961 gccacctccc acctaggaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa
     4021 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     4081 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     4141 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     4201 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     4261 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     4321 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     4381 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
     4441 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     4501 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     4561 tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacctcgag acctagaaaa
     4621 acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga
     4681 agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat
     4741 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
     4801 gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg
     4861 ctacttccct gattggcaga actacacacc agggccaggg atcagatatc cactgacctt
     4921 tggatggtgc tacaagctag taccagttga gcaagagaag gtagaagaag ccaatgaagg
     4981 agagaacacc cgcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga
     5041 agtattagag tggaggtttg acagccgcct agcatttcat cacatggccc gagagctgca
     5101 tccggactgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     5161 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     5221 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     5281 gaaaatctct agcagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
     5341 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     5401 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     5461 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     5521 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     5581 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     5641 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     5701 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     5761 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     5821 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     5881 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     5941 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     6001 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     6061 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     6121 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     6181 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     6241 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     6301 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     6361 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac
     6421 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
     6481 atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc
     6541 gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga cttcgccggt
     6601 gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt ggtgccggac
     6661 aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag
     6721 gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat cggcgagcag
     6781 ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca cttcgtggcc
     6841 gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg
     6901 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     6961 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     7021 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     7081 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     7141 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     7201 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     7261 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     7321 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     7381 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     7441 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     7501 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     7561 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     7621 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     7681 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     7741 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     7801 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     7861 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     7921 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     7981 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     8041 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     8101 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     8161 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     8221 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     8281 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     8341 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     8401 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     8461 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     8521 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     8581 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     8641 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     8701 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     8761 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     8821 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     8881 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     8941 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     9001 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     9061 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     9121 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     9181 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     9241 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     9301 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
     9361 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     9421 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
     9481 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata
     9541 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
     9601 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     9661 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     9721 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     9781 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
     9841 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     9901 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
     9961 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
    10021 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
    10081 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt
    10141 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
    10201 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
    10261 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
    10321 agcagtggcg cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac
    10381 gcaggactcg gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta
    10441 cgccaaaaat tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta
    10501 ttaagcgggg gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga
    10561 aaaaatataa attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta
    10621 atcctggcct gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat
    10681 cccttcagac aggatcagaa gaacttagat cattatataa tacagtagca accctctatt
    10741 gtgtgcatca aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag
    10801 agcaaaacaa aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga
    10861 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
    10921 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
    10981 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
    11041 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
    11101 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
    11161 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
    11221 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
    11281 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
    11341 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
    11401 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
    11461 tttaacataa ca
//
