LOCUS       sequence-242946-       11838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  donorAB-Neon targeting vector for human AAVS1 locus; knock-in of Dox
            controlled H2BGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11838)
  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 11838)
  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-H2BmNeon (donorAB-mNeon)
FEATURES             Location/Qualifiers
     source          1..11838
                     /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        2389..2692
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2694..2971
                     /label=chicken beta-actin promoter
     intron          2973..3990
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3914..3936
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3998..4017
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(4108..4725,4726..4731,4732..4767,4768..4770,4771..4806,
                     4807..4809,4810..4845,4846..4854)
                     /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    4864..4998
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4888..4907)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    5349..5430
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(5577..6287)
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      6023..6032
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      6284..6293
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(6306..6683)
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     promoter        complement(6712..7026)
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     primer_bind     complement(6715..6739)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    6786..6804
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6822..6840
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6857..6875
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6893..6911
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6929..6947
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    6964..6982
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    7000..7018
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    7074..7910
                     /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(7944..7963)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7945..7963)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7970..7987)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7970..7986)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7979..8001)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             8124..8426
                     /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     8305..8324
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(join(9823..10614,10615..10683))
                     /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     10446..10465
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      10807..11395
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     11296..11315
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     11549..11566
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    11683..11704
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(11719..11724,11725..11742,11743..11749)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11757..11773
                     /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     11762..11784
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     11781..11797
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11781..11797
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        11818..11836
                     /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 actaaaagga agtttttctg cagaaattga tgatctatta
     1861 aacaataaag atgtccacta aaatgaagtt ttttactgtc atactttgtt aagaagggtg
     1921 agaacagagt acctacattt tgaatggaag gattggagct acgggggtgg gggtggggtg
     1981 ggattagata aatgcctgct ctttactgaa ggctctttac tattgcttta tgataatgtt
     2041 tcatagttgg atatcataat ttaaacaagc aaaaccaaat taagggccag ctcattcctc
     2101 ccactcatga tctatagatc tatagatctc tcgtgggatc attgtttttc tcttgattcc
     2161 cactttgtgg ttctaagtac tgtggtttcc aaatgtgtca gtttcatagc ctgaagaacg
     2221 agatcagcag cctctgttcc acatacactt cattctcagt attgttttgc caagttctaa
     2281 ttccatcaga agctgggagc tccaccgcgg tggcggccgc tctagaacta gttattaata
     2341 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     2401 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     2461 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     2521 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     2581 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     2641 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt
     2701 gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta
     2761 tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg gggggcgcgc
     2821 gccaggcggg gcggggcggg gcgaggggcg gggcggggcg aggcggagag gtgcggcggc
     2881 agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg
     2941 gccctataaa aagcgaagcg cgcggcgggc ggggagtcgc tgcgacgctg ccttcgcccc
     3001 gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc
     3061 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa
     3121 tgacggcttg tttcttttct gtggctgcgt gaaagccttg aggggctccg ggagggccct
     3181 ttgtgcgggg ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg
     3241 tgcggctccg cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg
     3301 ctccgcagtg tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg cggggggggc
     3361 tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg
     3421 ggcgcgtcgg tcgggctgca accccccctg cacccccctc cccgagttgc tgagcacggc
     3481 ccggcttcgg gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg
     3541 gggtggcggc aggtgggggt gccgggcggg gcggggccgc ctcgggccgg ggagggctcg
     3601 ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc
     3661 cattgccttt tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt
     3721 gcggagccga aatctgggag gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg
     3781 tgcggcgccg gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt
     3841 ccccttctcc ctctccagcc tcggggctgt ccgcgggggg acggctgcct tcggggggga
     3901 cggggcaggg cggggttcgg cttctggcgt gtgaccggcg gctctagagc ctctgctaac
     3961 catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt tattgtgctg
     4021 tctcatcatt ttggcaaaga attcggcttg atcgaagcta gtggatcccc cgggctgcag
     4081 gaattcacca tgtctagact ggacaagatg tctagactgg acaagagcaa agtcataaac
     4141 ggcgctctgg aattactcaa tggagtcggt atcgaaggcc tgacgacaag gaaactcgct
     4201 caaaagctgg gagttgagca gcctaccctg tactggcacg tgaagaacaa gcgggccctg
     4261 ctcgatgccc tgccaatcga gatgctggac aggcatcata cccacttctg ccccctggaa
     4321 ggcgagtcat ggcaagactt tctgcggaac aacgccaagt cattccgctg tgctctcctc
     4381 tcacatcgcg acggggctaa agtgcatctc ggcacccgcc caacagagaa acagtacgaa
     4441 accctggaaa atcagctcgc gttcctgtgt cagcaaggct tctccctgga gaacgcactg
     4501 tacgctctgt ccgccgtggg ccactttaca ctgggctgcg tattggagga acaggagcat
     4561 caagtagcaa aagaggaaag agagacacct accaccgatt ctatgccccc acttctgaga
     4621 caagcaattg agctgttcga ccggcaggga gccgaacctg ccttcctttt cggcctggaa
     4681 ctaatcatat gtggcctgga gaaacagcta aagtgcgaaa gcggcgggcc ggccgacgcc
     4741 cttgacgatt ttgacttaga catgctccca gccgatgccc ttgacgactt tgaccttgat
     4801 atgctgcctg ctgacgctct tgacgatttt gaccttgaca tgctccccgg gtaactaagt
     4861 aaggatccag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     4921 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     4981 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     5041 gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtat ggctgattat
     5101 gatcctgcaa gcctcgtcgt cctggccgga ccacgctatc tgtgcaaggt ccccggcccc
     5161 ggacgcgcgc tccatgagca gagcgcccgc cgccgaggcg aagactcggg cggcgccctg
     5221 cccgtcccac caggtcaaca ggcggtaacc ggcctcttca tcgggaatgc gcgcgacctt
     5281 cagcatcgcc ggcatgtccc cctggcggac gggaagtatc cagctcgacc aagcttagta
     5341 ctgtcgaggc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     5401 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     5461 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     5521 atggctgatt atgatcctct agagatatcg tcgacaagct tatcgatgcg gccgctttac
     5581 ttgtacagct cgtccatgcc catcacatcg gtaaaggcct tttgccactc cttgaagttg
     5641 agctcggtct tggagtgctt gagctccgtc ttacggaaca cgtacatcgg ctggttcttc
     5701 agatagttag ccgccattgg cttggcaaag gtgtaggtgg tccgcgcagt gctccggtag
     5761 cgcttgccat ttccagtggt gtaactccac ttaaaggtac tgatgatggt tttgtcgttg
     5821 gggtaagtct tcttcgacct gcaccagtcc gcagcggtca gcgagttggt catcacagga
     5881 ccgtcagcag ggaaaccagt ccccttcacc tgggcctctc ctttgatgtg gcttccctcg
     5941 taggtgtagc ggtagttaac agtaagggag gcaccatctt caaactgcat tgtgcgatgg
     6001 acttggtatc cggagccatc taccatggcg gcctggaaag gcgacatccc gtcagggtag
     6061 ggcaggtact gatggaagcc atacccgata tgagggacca gaatccaggg ggagaactgg
     6121 aggtcaccct tggtggactt caggtttaac tcctcataac catcatttgg attgccggtg
     6181 ccctgaccca ccatgtcaaa gtccacaccg ttgatggagc caaagatgtg taactcatgt
     6241 gtcgctggga gagaggccat gttatcctcc tcgcccttgc tcaccatggt ggcgaccggt
     6301 ggatccttag cgctggtgta cttggtgatg gccttagtac cctcggacac ggcgtgcttg
     6361 gccaactccc caggcagcag caggcgcacg gccgtctgga tctccctgga ggtgatggtc
     6421 gagcgcttgt tgtaatgcgc caggcgggaa gcctcacctg cgatgcgctc gaaaatgtcg
     6481 ttcacaaacg aattcatgat gcccatggcc ttggacgaaa tgccggtgtc agggtggacc
     6541 tgcttcagaa ccttgtacac atagatggaa tagctctcct tgcggctgcg cttgcgcttc
     6601 ttgccgcctt tcttctgcgc cttagtcacc gccttcttgg agcccttttt cggggcggga
     6661 gcagacttcg ctggctctgg catggtggcg gtaccgagct cgaattctcc aggcgatctg
     6721 acggttcact aaacgagctc tgcttatata ggcctcccac cgtacacgcc tacctcgaca
     6781 tacgttctct atcactgata gggagtaaac tcgacatacg ttctctatca ctgataggga
     6841 taaactcgac atacgttctc tatcactgat agggagtaaa ctcgacatac gttctctatc
     6901 actgataggg agtaaactcg acatacgttc tctatcactg atagggagta aactcgacat
     6961 cgttctctat cactgatagg gagtaaactc gacatacgtt ctctatcact gatagggagt
     7021 aaactcgacc catagagccc accgcatccc cagcatgcct gctattaagc tttactaggg
     7081 acaggattgg tgacagaaaa gccccatcct taggcctcct ccttcctagt ctcctgatat
     7141 tgggtctaac ccccacctcc tgttaggcag attccttatc tggtgacaca cccccatttc
     7201 ctggagccat ctctctcctt gccagaacct ctaaggtttg cttacgatgg agccagagag
     7261 gatcctggga gggagagctt ggcagggggt gggagggaag ggggggatgc gtgacctgcc
     7321 cggttctcag tggccaccct gcgctaccct ctcccagaac ctgagctgct ctgacgcggc
     7381 tgtctggtgc gtttcactga tcctggtgct gcagcttcct tacacttccc aagaggagaa
     7441 gcagtttgga aaaacaaaat cagaataagt tggtcctgag ttctaacttt ggctcttcac
     7501 ctttctagtc cccaatttat attgttcctc cgtgcgtcag ttttacctgt gagataaggc
     7561 cagtagccag ccccgtcctg gcagggctgt ggtgaggagg ggggtgtccg tgtggaaaac
     7621 tccctttgtg agaatggtgc gtcctaggtg ttcaccaggt cgtggccgcc tctactccct
     7681 ttctctttct ccatccttct ttccttaaag agtccccagt gctatctggg acatattcct
     7741 ccgcccagag cagggtcccg cttccctaag gccctgctct gggcttctgg gtttgagtcc
     7801 ttggcaagcc caggagaggc gctcaggctt ccctgtcccc cttcctcgtc caccatctca
     7861 tgcccctggc tctcctgccc cttccctaca ggggttcctg gctctgctct aagggcgaat
     7921 tcgcggccgc taaattcaat tcgccctata gtgagtcgta ttacaattca ctggccgtcg
     7981 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     8041 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     8101 agttgcgcag cctatacgta cggcagttta aggtttacac ctataaaaga gagagccgtt
     8161 atcgtctgtt tgtggatgta cagagtgata ttattgacac gccggggcga cggatggtga
     8221 tccccctggc cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg
     8281 tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga tatggccagt gtgccggtct
     8341 ccgttatcgg ggaagaagtg gctgatctca gccaccgcga aaatgacatc aaaaacgcca
     8401 ttaacctgat gttctgggga atataaatgt caggcatgag attatcaaaa aggatcttca
     8461 cctagatcct tttcacgtag aaagccagtc cgcagaaacg gtgctgaccc cggatgaatg
     8521 tcagctactg ggctatctgg acaagggaaa acgcaagcgc aaagagaaag caggtagctt
     8581 gcagtgggct tacatggcga tagctagact gggcggtttt atggacagca agcgaaccgg
     8641 aattgccagc tggggcgccc tctggtaagg ttgggaagcc ctgcaaagta aactggatgg
     8701 ctttcttgcc gccaaggatc tgatggcgca ggggatcaag ctctgatcaa gagacaggat
     8761 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     8821 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     8881 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     8941 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     9001 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     9061 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     9121 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     9181 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     9241 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     9301 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgc atggcgatgc ctgcttgccg
     9361 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     9421 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     9481 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     9541 gccttctatc gccttcttga cgagttcttc tgaattatta acgcttacaa tttcctgatg
     9601 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatcaggtg gcacttttcg
     9661 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     9721 gctcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     9781 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     9841 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     9901 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     9961 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
    10021 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
    10081 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
    10141 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
    10201 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
    10261 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
    10321 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgaatactc
    10381 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
    10441 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
    10501 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
    10561 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
    10621 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
    10681 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgaccaa
    10741 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
    10801 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
    10861 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
    10921 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
    10981 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
    11041 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
    11101 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
    11161 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
    11221 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
    11281 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
    11341 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
    11401 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
    11461 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
    11521 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
    11581 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
    11641 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
    11701 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
    11761 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agctcagaat
    11821 taaccctcac taaaggga
//
