LOCUS       sequence-363155-        8192 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor template for knock-in of p2a-CreERT2 into the medaka col10a1 
            locus via CRISPR/Cas9-mediated homology directed repair.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8192)
  AUTHORS   Hui TW, Winkler C
  TITLE     A novel non-disruptive and efficient knock-in allows fate tracing of
            resident osteoblast progenitors during repair of vertebral lesions 
            in medaka.
  JOURNAL   Development. 2022 May 20. pii: 275483. doi: 10.1242/dev.200238.
  PUBMED    35593425
REFERENCE   2  (bases 1 to 8192)
  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: 
            "Development. 2022 May 20. pii: 275483. doi: 10.1242/dev.200238."
COMMENT     Sequence Label: pTOPO-col10a1-KI-donor
FEATURES             Location/Qualifiers
     source          1..8192
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(908..1282)
                     /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"
     regulatory      1279..1288
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(1488..2282)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(1488..2282)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1599..1618)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2209..2228
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(2631..2933)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(2733..2752)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     3056..3078
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3070..3087
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3071..3087
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3094..3112
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3521..3577
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3578..4606
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     CDS             4619..5554
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             4667..5560
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATA"
     polyA_signal    5640..5721
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     5932..5951
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(6023..6736,6737..6739,6740..6742))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(6026..6736,6737..6739,6740..6742))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6068..6089)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     6417..6436
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     promoter        complement(7936..7954)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(7937..7954)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(7972..7988)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7972..7988)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7985..8007)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7996..8012
                     /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(8020..8026,8027..8044,8045..8050))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8065..8086
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 gcacgtgtca gtcctgctcc tcggccacga agtgcacgca gttgccggcc gggtcgcgca
      961 gggcgaactc ccgcccccac ggctgctcgc cgatctcggt catggccggc ccggaggcgt
     1021 cccggaagtt cgtggacacg acctccgacc actcggcgta cagctcgtcc aggccgcgca
     1081 cccacaccca ggccagggtg ttgtccggca ccacctggtc ctggaccgcg ctgatgaaca
     1141 gggtcacgtc gtcccggacc acaccggcga agtcgtcctc cacgaagtcc cgggagaacc
     1201 cgagccggtc ggtccagaac tcgaccgctc cggcgacgtc gcgcgcggtg agcaccggaa
     1261 cggcactggt caacttggcc atggtggccc tcctcacgtg ctattattga agcatttatc
     1321 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     1381 gggttccgcg cacatttccc cgaaaagtgc cacctgatgc ggtgtgaaat accgcacaga
     1441 tgcgtaagga gaaaataccg catcaggaaa ttgtaagcgt taataattca gaagaactcg
     1501 tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg
     1561 aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct
     1621 atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg
     1681 ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg
     1741 ccgtcgggca tgctcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc
     1801 tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg
     1861 atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc
     1921 cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga
     1981 tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg
     2041 agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct
     2101 tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc
     2161 gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag
     2221 ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc
     2281 atgcgaaacg atcctcatcc tgtctcttga tcagagcttg atcccctgcg ccatcagatc
     2341 cttggcggca agaaagccat ccagtttact ttgcagggct tcccaacctt accagagggc
     2401 gccccagctg gcaattccgg ttcgcttgct gtccataaaa ccgcccagtc tagctatcgc
     2461 catgtaagcc cactgcaagc tacctgcttt ctctttgcgc ttgcgttttc ccttgtccag
     2521 atagcccagt agctgacatt catccggggt cagcaccgtt tctgcggact ggctttctac
     2581 gtgaaaagga tctaggtgaa gatccttttt gataatctca tgcctgacat ttatattccc
     2641 cagaacatca ggttaatggc gtttttgatg tcattttcgc ggtggctgag atcagccact
     2701 tcttccccga taacggagac cggcacactg gccatatcgg tggtcatcat gcgccagctt
     2761 tcatccccga tatgcaccac cgggtaaagt tcacgggaga ctttatctga cagcagacgt
     2821 gcactggcca gggggatcac catccgtcgc cccggcgtgt caataatatc actctgtaca
     2881 tccacaaaca gacgataacg gctctctctt ttataggtgt aaaccttaaa ctgccgtacg
     2941 tataggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     3001 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     3061 tcacgacgtt gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattg
     3121 ggccctctag atgcatgctc gagcggccgc cagtgtgatg gatatctgca gaattcgccc
     3181 ttcgaccacc tctggaaccg cagcagcctc tgctgtttca ccccatagcc ctgagaccgg
     3241 catcttcact tgccaagttc caggtgttta ttatttctct tacggtatcc atgtaaatgg
     3301 aaatcatgcc ctggtggctt tgtacaagaa tggtcagccc attgtgttca cctatagtga
     3361 gtacaacaaa ggcttcctgg accagatgtc tggcagtgct gtcctcttgc ttgatgtgca
     3421 ggacacagtc tacatccaga tccctgatga tgaggccaat ggtgtctttg ctgctgagaa
     3481 tgtacattgt acgttttccg gttttctcat cgcttcaacg gctacaaact tcagcctgct
     3541 gaagcaggct ggagacgtgg aggagaaccc tggacctatg tccaatttac taaccgtaca
     3601 ccaaaatttg cctgcattac cggtcgatgc aacgagtgat gaggttcgca agaacctgat
     3661 ggacatgttc agggatcgcc aggcgttttc tgagcatacc tggaaaatgc ttctgtccgt
     3721 ttgccggtcg tgggcggcat ggtgcaagtt gaataaccgg aaatggtttc ccgcagaacc
     3781 tgaagatgtt cgcgattatc ttctatatct tcaggcgcgc ggtctggcag taaaaactat
     3841 ccagcaacat ttgggccagc taaacatgct tcatcgtcgg tccgggctgc cacgaccaag
     3901 tgacagcaat gctgtttcac tggttatgcg gcggatccga aaagaaaacg ttgatgccgg
     3961 tgaacgtgca aaacaggctc tagcgttcga acgcactgat ttcgaccagg ttcgttcact
     4021 catggaaaat agcgatcgct gccaggatat acgtaatctg gcatttctgg ggattgctta
     4081 taacaccctg ttacgtatag ccgaaattgc caggatcagg gttaaagata tctcacgtac
     4141 tgacggtggg agaatgttaa tccatattgg cagaacgaaa acgctggtta gcaccgcagg
     4201 tgtagagaag gcacttagcc tgggggtaac taaactggtc gagcgatgga tttccgtctc
     4261 tggtgtagct gatgatccga ataactacct gttttgccgg gtcagaaaaa atggtgttgc
     4321 cgcgccatct gccaccagcc agctatcaac tcgcgccctg gaagggattt ttgaagcaac
     4381 tcatcgattg atttacggcg ctaaggatga ctctggtcag agatacctgg cctggtctgg
     4441 acacagtgcc cgtgtcggag ccgcgcgaga tatggcccgc gctggagttt caataccgga
     4501 gatcatgcaa gctggtggct ggaccaatgt aaatattgtc atgaactata tccgtaacct
     4561 ggatagtgaa acaggggcaa tggtgcgcct gctggaagat ggcgatctcg agccatctgc
     4621 tggagacatg agagctgcca acctttggcc aagcccgctc atgatcaaac gctctaagaa
     4681 gaacagcctg gccttgtccc tgacggccga ccagatggtc agtgccttgt tggatgctga
     4741 gccccccata ctctattccg agtatgatcc taccagaccc ttcagtgaag cttcgatgat
     4801 gggcttactg accaacctgg cagacaggga gctggttcac atgatcaact gggcgaagag
     4861 ggtgccaggc tttgtggatt tgaccctcca tgatcaggtc caccttctag aatgtgcctg
     4921 gctagagatc ctgatgattg gtctcgtctg gcgctccatg gagcacccag tgaagctact
     4981 gtttgctcct aacttgctct tggacaggaa ccagggaaaa tgtgtagagg gcatggtgga
     5041 gatcttcgac atgctgctgg ctacatcatc tcggttccgc atgatgaatc tgcagggaga
     5101 ggagtttgtg tgcctcaaat ctattatttt gcttaattct ggagtgtaca catttctgtc
     5161 cagcaccctg aagtctctgg aagagaagga ccatatccac cgagtcctgg acaagatcac
     5221 agacactttg atccacctga tggccaaggc aggcctgacc ctgcagcagc agcaccagcg
     5281 gctggcccag ctcctcctca tcctctccca catcaggcac atgagtaaca aaggcatgga
     5341 gcatctgtac agcatgaagt gcaagaacgt ggtgcccctc tatgacctgc tgctggaggc
     5401 ggcggacgcc caccgcctac atgcgcccac tagccgtgga ggggcatccg tggaggagac
     5461 ggaccaaagc cacttggcca ctgcgggctc tacttcatcg cattccttgc aaaagtatta
     5521 catcacgggg gaggcagagg gtttccctgc cacagcttga taggttgatc acgcggcagg
     5581 ttccaaaagc caaagctagc aagggcgaat tccagcacac tggcggccgt tactagaatg
     5641 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     5701 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     5761 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tatggctgat
     5821 tatgatcctc tagatcagat ctcttgttta ttgcagctta taatggttac aaataaagca
     5881 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     5941 ccaaactcat caatgtatct tatcatgtct ggatctacgt aatacgactc actatagttc
     6001 tagaggctcg agaggggccg ctttacttgt acagctcgtc catgccgaga gtgatcccgg
     6061 cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg tctttgctca
     6121 gggcggactg ggtgctcagg tagtggttgt cgggcagcag cacggggccg tcgccgatgg
     6181 gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc gtcctcgatg ttgtggcgga
     6241 tcttgaagtt caccttgatg ccgttcttct gcttgtcggc catgatatag acgttgtggc
     6301 tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg aagtcgatgc
     6361 ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg gcgcgggtct
     6421 tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct tcgggcatgg
     6481 cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcggctgaag cactgcacgc
     6541 cgtaggtcag ggtggtcacg agggtgggcc agggcacggg cagcttgccg gtggtgcaga
     6601 tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg gacacgctga
     6661 acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc ccggtgaaca
     6721 gctcctcgcc cttgctcacc atggtcactg tctgctttgc tgttggtctg ggctcctggg
     6781 tcactggctt actaatggag tctttatgta tgaggactct tatcaattgt tcttctataa
     6841 aggtctgcag tgtttctgtt cgtcccctac atggacaccc agagcctcct aaatacagga
     6901 gccctgataa ctgcacaagt gctcagattc cagcagggtg gaaaatgaga taaagtgtgc
     6961 agatggggag ggggacgtga atgagagatt tgagggatga aagggatgga tgaacgcatt
     7021 gaaaatagcc cctttcacaa aataatacca gtaaattgtc atataattaa tgaccagacc
     7081 tttacctgta aatgatgtga tgttcatata cagaagaatg ccaacaattt acagatgatt
     7141 ttacaaattc ttaattttcc ttttctaaat ggatttgttc acacattatc tcttcaaagc
     7201 aattgaattt tctggaaaag attggctgtg tgtgtaaaag gtgctactga ggaatttaat
     7261 gtgacatgga atgaagcaaa cagcagccta agacaggatg gcaggaaaaa tgtcacatgt
     7321 ttaaaataag agtgaaacca acgacctgga cttatagaga ctgtgtgtcc ttaggcaaag
     7381 ctcttgtaca aacagtgctt gtgactgtga aattcggggt ttgcctggat tgtgttaaat
     7441 gtgtgcttgg atgtatcata aaaaagaatc ggttctctgt atttgacatt ttctgtaatt
     7501 tatctaaagt tacaactggc tatgcctgat ttttatcact attgaatcaa tcattttaaa
     7561 gaatgttaag tctgctgatt tggagagctc atttacattt attttgaatg tctcttattt
     7621 aacacaactg atttaagctt ttaggttgat cacgcatcag gttccaaaag ccaaaccaac
     7681 taattttgca atctattcct caaagtaaac tctctgaccc tccctattct ccaatagaca
     7741 ggcagttctt taaggaatag tagcatctca acttgaaaac tacaagaaaa aggtgctcaa
     7801 cagaaggaaa actaatttat gaaagcaaat gatcagggat ggggggaagc aaacaagagc
     7861 acttttttgc cgttccggct gctgcggttc cagaggtgga gtgagggtac caagcttgat
     7921 gcatagcttg agtattctat agtgtcacct aaatagcttg gcgtaatcat ggtcatagct
     7981 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     8041 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     8101 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     8161 cgcggggaga ggcggtttgc gtattgggcg ct
//
