LOCUS       sequence-169633-       11846 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  donorAB targeting vector for human AAVS1 locus; knock-in of Dox 
            controlled H2BGFP expression system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11846)
  AUTHORS   Tejero R, Huang Y, Katsyv I, Kluge M, Lin JY, Tome-Garcia J, Daviaud
            N, Wang Y, Zhang B, Tsankova NM, Friedel CC, Zou H, Friedel RH
  TITLE     Gene signatures of quiescent glioblastoma cells reveal mesenchymal 
            shift and interactions with niche microenvironment.
  JOURNAL   EBioMedicine. 2019 Apr 2. pii: S2352-3964(19)30209-9. doi: 
            10.1016/j.ebiom.2019.03.064.
  PUBMED    30952620
REFERENCE   2  (bases 1 to 11846)
  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: 
            "EBioMedicine. 2019 Apr 2. pii: S2352-3964(19)30209-9. doi: 
            10.1016/j.ebiom.2019.03.064."
COMMENT     Sequence Label: pAAVS1-Neo-CAG-M2rtTA-H2BGFP (donor AB)
FEATURES             Location/Qualifiers
     source          1..11846
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    33..836
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     misc_feature    843..868
                     /label=SA
                     /note="splice acceptor site"
     CDS             892..945
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             961..1764
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     enhancer        2390..2693
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2695..2972
                     /label=chicken beta-actin promoter
     intron          2973..3989
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3913..3935
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3997..4016
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(4107..4724,4725..4730,4731..4766,4767..4769,4770..4805,
                     4806..4808,4809..4844,4845..4853)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     polyA_signal    4863..4997
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4887..4906)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    5348..5429
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(join(5576..6289,6290..6292,6293..6295))
                     /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(5579..6289,6290..6292,6293..6295))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5621..5642)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     5970..5989
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6229..6250
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      6292..6301
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(6314..6691)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     promoter        complement(6720..7034)
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     primer_bind     complement(6723..6747)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    6794..6812
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6830..6848
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6865..6883
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6901..6919
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6937..6955
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6972..6990
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7008..7026
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    7082..7918
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     primer_bind     complement(7952..7971)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7953..7971)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7978..7995)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7978..7994)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7987..8009)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             8132..8434
                     /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     8313..8332
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(join(9831..10622,10623..10691))
                     /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     10454..10473
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10815..11403
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11304..11323
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11557..11574
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11691..11712
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11727..11732,11733..11750,11751..11757)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11765..11781
                     /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)."
     primer_bind     11770..11792
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11789..11805
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11789..11805
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        11826..11844
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ctagtcctgc aggtttaaac gaattcgccc tttgctttct ctgaccagca ttctctcccc
       61 tgggcctgtg ccgctttctg tctgcagctt gtggcctggg tcacctctac ggctggccca
      121 gatccttccc tgccgcctcc ttcaggttcc gtcttcctcc actccctctt ccccttgctc
      181 tctgctgtgt tgctgcccaa ggatgctctt tccggagcac ttccttctcg gcgctgcacc
      241 acgtgatgtc ctctgagcgg atcctccccg tgtctgggtc ctctccgggc atctctcctc
      301 cctcacccaa ccccatgccg tcttcactcg ctgggttccc ttttccttct ccttctgggg
      361 cctgtgccat ctctcgtttc ttaggatggc cttctccgac ggatgtctcc cttgcgtccc
      421 gcctcccctt cttgtaggcc tgcatcatca ccgtttttct ggacaacccc aaagtacccc
      481 gtctccctgg ctttagccac ctctccatcc tcttgctttc tttgcctgga caccccgttc
      541 tcctgtggat tcgggtcacc tctcactcct ttcatttggg cagctcccct acccccctta
      601 cctctctagt ctgtgctagc tcttccagcc ccctgtcatg gcatcttcca ggggtccgag
      661 agctcagcta gtcttcttcc tccaacccgg gcccctatgt ccacttcagg acagcatgtt
      721 tgctgcctcc agggatcctg tgtccccgag ctgggaccac cttatattcc cagggccggt
      781 taatgtggct ctggttctgg gtacttttat ctgtcccctc caccccacag tggggcaagc
      841 ttctgacctc ttctcttcct cccacagggc ctcgagagat ctggcagcgg agagggcaga
      901 ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctaggct cgagacgcgt
      961 atgggatcgg ccattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     1021 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     1081 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     1141 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     1201 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     1261 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     1321 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     1381 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     1441 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     1501 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     1561 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     1621 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     1681 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     1741 cgccttcttg acgagttctt ctgaggggat ccgctgtaag tctgcagaaa ttgatgatct
     1801 attaaacaat aaagatgtcc actaaaatgg aagtttttcc tgtcatactt tgttaagaag
     1861 ggtgagaaca gatgtccact aaaatggaag tttttcctgt catactttgt taagaagggt
     1921 gagaacagag tacctacatt ttgaatggaa ggattggagc tacgggggtg ggggtggggt
     1981 gggattagat aaatgcctgc tctttactga aggctcttta ctattgcttt atgataatgt
     2041 ttcatagttg gatatcataa tttaaacaag caaaaccaaa ttaagggcca gctcattcct
     2101 cccactcatg atctatagat ctatagatct ctcgtgggat cattgttttt ctcttgattc
     2161 ccactttgtg gttctaagta ctgtggtttc caaatgtgtc agtttcatag cctgaagaac
     2221 gagatcagca gcctctgttc cacatacact tcattctcag tattgttttg ccaagttcta
     2281 attccatcag aagctgggag ctccaccgcg gtggcggccg ctctagaact agttattaat
     2341 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     2401 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     2461 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     2521 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
     2581 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
     2641 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtcgagg
     2701 tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt
     2761 atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg ggggggcgcg
     2821 cgccaggcgg ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg
     2881 cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc
     2941 ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg
     3001 tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc
     3061 cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagcgc ttggtttaat
     3121 gacggcttgt ttcttttctg tggctgcgtg aaagccttga ggggctccgg gagggccctt
     3181 tgtgcggggg gagcggctcg gggggtgcgt gcgtgtgtgt gtgcgtgggg agcgccgcgt
     3241 gcggctccgc gctgcccggc ggctgtgagc gctgcgggcg cggcgcgggg ctttgtgcgc
     3301 tccgcagtgt gcgcgagggg agcgcggccg ggggcggtgc cccgcggtgc ggggggggct
     3361 gcgaggggaa caaaggctgc gtgcggggtg tgtgcgtggg ggggtgagca gggggtgtgg
     3421 gcgcgtcggt cgggctgcaa ccccccctgc acccccctcc ccgagttgct gagcacggcc
     3481 cggcttcggg tgcggggctc cgtacggggc gtggcgcggg gctcgccgtg ccgggcgggg
     3541 ggtggcggca ggtgggggtg ccgggcgggg cggggccgcc tcgggccggg gagggctcgg
     3601 gggaggggcg cggcggcccc cggagcgccg gcggctgtcg aggcgcggcg agccgcagcc
     3661 attgcctttt atggtaatcg tgcgagaggg cgcagggact tcctttgtcc caaatctgtg
     3721 cggagccgaa atctgggagg cgccgccgca ccccctctag cgggcgcggg gcgaagcggt
     3781 gcggcgccgg caggaaggaa atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc
     3841 cccttctccc tctccagcct cggggctgtc cgcgggggga cggctgcctt cgggggggac
     3901 ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc
     3961 atgttcatgc cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt
     4021 ctcatcattt tggcaaagaa ttcggcttga tcgaagctag tggatccccc gggctgcagg
     4081 aattcaccat gtctagactg gacaagatgt ctagactgga caagagcaaa gtcataaacg
     4141 gcgctctgga attactcaat ggagtcggta tcgaaggcct gacgacaagg aaactcgctc
     4201 aaaagctggg agttgagcag cctaccctgt actggcacgt gaagaacaag cgggccctgc
     4261 tcgatgccct gccaatcgag atgctggaca ggcatcatac ccacttctgc cccctggaag
     4321 gcgagtcatg gcaagacttt ctgcggaaca acgccaagtc attccgctgt gctctcctct
     4381 cacatcgcga cggggctaaa gtgcatctcg gcacccgccc aacagagaaa cagtacgaaa
     4441 ccctggaaaa tcagctcgcg ttcctgtgtc agcaaggctt ctccctggag aacgcactgt
     4501 acgctctgtc cgccgtgggc cactttacac tgggctgcgt attggaggaa caggagcatc
     4561 aagtagcaaa agaggaaaga gagacaccta ccaccgattc tatgccccca cttctgagac
     4621 aagcaattga gctgttcgac cggcagggag ccgaacctgc cttccttttc ggcctggaac
     4681 taatcatatg tggcctggag aaacagctaa agtgcgaaag cggcgggccg gccgacgccc
     4741 ttgacgattt tgacttagac atgctcccag ccgatgccct tgacgacttt gaccttgata
     4801 tgctgcctgc tgacgctctt gacgattttg accttgacat gctccccggg taactaagta
     4861 aggatccaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     4921 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     4981 gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg
     5041 aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg gctgattatg
     5101 atcctgcaag cctcgtcgtc ctggccggac cacgctatct gtgcaaggtc cccggccccg
     5161 gacgcgcgct ccatgagcag agcgcccgcc gccgaggcga agactcgggc ggcgccctgc
     5221 ccgtcccacc aggtcaacag gcggtaaccg gcctcttcat cgggaatgcg cgcgaccttc
     5281 agcatcgccg gcatgtcccc ctggcggacg ggaagtatcc agctcgacca agcttagtac
     5341 tgtcgaggca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     5401 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     5461 aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta aaacctctac aaatgtggta
     5521 tggctgatta tgatcctcta gagatatcgt cgacaagctt atcgatgcgg ccgctttact
     5581 tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca
     5641 tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga ctgggtgctc aggtagtggt
     5701 tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga
     5761 gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg atgccgttct
     5821 tctgcttgtc ggccatgata tagacgttgt ggctgttgta gttgtactcc agcttgtgcc
     5881 ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg
     5941 tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga
     6001 tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca
     6061 tgtggtcggg gtagcggctg aagcactgca cgccgtaggt cagggtggtc acgagggtgg
     6121 gccagggcac gggcagcttg ccggtggtgc agatgaactt cagggtcagc ttgccgtagg
     6181 tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca
     6241 gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc accatggtgg
     6301 cgaccggtgg atccttagcg ctggtgtact tggtgatggc cttagtaccc tcggacacgg
     6361 cgtgcttggc caactcccca ggcagcagca ggcgcacggc cgtctggatc tccctggagg
     6421 tgatggtcga gcgcttgttg taatgcgcca ggcgggaagc ctcacctgcg atgcgctcga
     6481 aaatgtcgtt cacaaacgaa ttcatgatgc ccatggcctt ggacgaaatg ccggtgtcag
     6541 ggtggacctg cttcagaacc ttgtacacat agatggaata gctctccttg cggctgcgct
     6601 tgcgcttctt gccgcctttc ttctgcgcct tagtcaccgc cttcttggag ccctttttcg
     6661 gggcgggagc agacttcgct ggctctggca tggtggcggt accgagctcg aattctccag
     6721 gcgatctgac ggttcactaa acgagctctg cttatatagg cctcccaccg tacacgccta
     6781 cctcgacata cgttctctat cactgatagg gagtaaactc gacatacgtt ctctatcact
     6841 gatagggata aactcgacat acgttctcta tcactgatag ggagtaaact cgacatacgt
     6901 tctctatcac tgatagggag taaactcgac atacgttctc tatcactgat agggagtaaa
     6961 ctcgacatcg ttctctatca ctgataggga gtaaactcga catacgttct ctatcactga
     7021 tagggagtaa actcgaccca tagagcccac cgcatcccca gcatgcctgc tattaagctt
     7081 tactagggac aggattggtg acagaaaagc cccatcctta ggcctcctcc ttcctagtct
     7141 cctgatattg ggtctaaccc ccacctcctg ttaggcagat tccttatctg gtgacacacc
     7201 cccatttcct ggagccatct ctctccttgc cagaacctct aaggtttgct tacgatggag
     7261 ccagagagga tcctgggagg gagagcttgg cagggggtgg gagggaaggg ggggatgcgt
     7321 gacctgcccg gttctcagtg gccaccctgc gctaccctct cccagaacct gagctgctct
     7381 gacgcggctg tctggtgcgt ttcactgatc ctggtgctgc agcttcctta cacttcccaa
     7441 gaggagaagc agtttggaaa aacaaaatca gaataagttg gtcctgagtt ctaactttgg
     7501 ctcttcacct ttctagtccc caatttatat tgttcctccg tgcgtcagtt ttacctgtga
     7561 gataaggcca gtagccagcc ccgtcctggc agggctgtgg tgaggagggg ggtgtccgtg
     7621 tggaaaactc cctttgtgag aatggtgcgt cctaggtgtt caccaggtcg tggccgcctc
     7681 tactcccttt ctctttctcc atccttcttt ccttaaagag tccccagtgc tatctgggac
     7741 atattcctcc gcccagagca gggtcccgct tccctaaggc cctgctctgg gcttctgggt
     7801 ttgagtcctt ggcaagccca ggagaggcgc tcaggcttcc ctgtccccct tcctcgtcca
     7861 ccatctcatg cccctggctc tcctgcccct tccctacagg ggttcctggc tctgctctaa
     7921 gggcgaattc gcggccgcta aattcaattc gccctatagt gagtcgtatt acaattcact
     7981 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     8041 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     8101 ttcccaacag ttgcgcagcc tatacgtacg gcagtttaag gtttacacct ataaaagaga
     8161 gagccgttat cgtctgtttg tggatgtaca gagtgatatt attgacacgc cggggcgacg
     8221 gatggtgatc cccctggcca gtgcacgtct gctgtcagat aaagtctccc gtgaacttta
     8281 cccggtggtg catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt
     8341 gccggtctcc gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa
     8401 aaacgccatt aacctgatgt tctggggaat ataaatgtca ggcatgagat tatcaaaaag
     8461 gatcttcacc tagatccttt tcacgtagaa agccagtccg cagaaacggt gctgaccccg
     8521 gatgaatgtc agctactggg ctatctggac aagggaaaac gcaagcgcaa agagaaagca
     8581 ggtagcttgc agtgggctta catggcgata gctagactgg gcggttttat ggacagcaag
     8641 cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt gggaagccct gcaaagtaaa
     8701 ctggatggct ttcttgccgc caaggatctg atggcgcagg ggatcaagct ctgatcaaga
     8761 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     8821 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     8881 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     8941 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     9001 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     9061 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     9121 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     9181 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     9241 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     9301 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatgcat ggcgatgcct
     9361 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     9421 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     9481 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     9541 agcgcatcgc cttctatcgc cttcttgacg agttcttctg aattattaac gcttacaatt
     9601 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atcaggtggc
     9661 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     9721 atgtatccgc tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     9781 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     9841 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     9901 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     9961 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
    10021 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
    10081 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
    10141 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
    10201 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
    10261 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
    10321 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
    10381 gaatactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
    10441 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
    10501 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
    10561 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
    10621 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
    10681 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
    10741 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
    10801 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
    10861 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
    10921 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
    10981 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
    11041 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
    11101 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
    11161 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
    11221 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
    11281 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
    11341 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
    11401 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
    11461 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
    11521 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
    11581 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
    11641 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
    11701 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
    11761 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
    11821 ctcagaatta accctcacta aaggga
//
