LOCUS       sequence-147885-       12744 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAVS1 targeting donor plasmid with Flpe-ERT2 expression cassette and
            Neo selection gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12744)
  AUTHORS   Chen Y, Cao J, Xiong M, Petersen AJ, Dong Y, Tao Y, Huang CT, Du Z, 
            Zhang SC
  TITLE     Engineering Human Stem Cell Lines with Inducible Gene Knockout using
            CRISPR/Cas9.
  JOURNAL   Cell Stem Cell. 2015 Jul 1. pii: S1934-5909(15)00261-1. doi: 
            10.1016/j.stem.2015.06.001.
  PUBMED    26145478
REFERENCE   2  (bases 1 to 12744)
  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: "Cell Stem 
            Cell. 2015 Jul 1. pii: S1934-5909(15)00261-1. doi: 
            10.1016/j.stem.2015.06.001."
COMMENT     Sequence Label: AAVS1-Neo-CAG-Flpe-ERT2
FEATURES             Location/Qualifiers
     source          1..12744
                     /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             949..1752
                     /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
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1784..1801)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1790..2014
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        2082..2461
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2463..2740
                     /label=chicken beta-actin promoter
     intron          2741..3757
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3681..3703
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3765..3784
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      3821..3830
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5126..6061
                     /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             5174..6067
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATA"
     misc_feature    6101..6689
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6154..6174)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6572..6583)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6721..7197
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(6920..6939)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(7307..7326)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7372..7427
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7426..7445)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7788..7804)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7812..7828
                     /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(7836..7842,7843..7860,7861..7866))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7881..7902
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    7976..8812
                     /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(8846..8865)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8847..8865)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8872..8889)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8872..8888)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8881..8903)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             9026..9328
                     /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     9207..9226
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(join(10725..11516,11517..11585))
                     /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     11348..11367
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11709..12297
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12198..12217
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     12451..12468
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    12585..12606
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(12621..12626,12627..12644,12645..12651)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12659..12675
                     /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     12683..12699
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        12720..12738
                     /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 gccctaggat gggatcggcc
      961 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     1021 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     1081 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     1141 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     1201 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     1261 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     1321 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     1381 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     1441 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     1501 gatgatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     1561 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     1621 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     1681 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     1741 gagttcttct gaggggatca attctctaga gctcgctgat cagcctcgac tgtgccttct
     1801 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     1861 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     1921 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     1981 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     2041 ggctcgacta gagcttgcgg aacccttaat ggtaccgtct cgacattgat tattgactag
     2101 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     2161 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     2221 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     2281 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     2341 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     2401 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     2461 ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc
     2521 aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggggg
     2581 ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg ggcggggcga ggcggagagg
     2641 tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg
     2701 gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct
     2761 tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc
     2821 gttactccca caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt
     2881 ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa agccttgagg ggctccggga
     2941 gggccctttg tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag
     3001 cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct
     3061 ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc cgcggtgcgg
     3121 ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg ggtgagcagg
     3181 gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc gagttgctga
     3241 gcacggcccg gcttcgggtg cggggctccg tacggggcgt ggcgcggggc tcgccgtgcc
     3301 gggcgggggg tggcggcagg tgggggtgcc gggcggggcg gggccgcctc gggccgggga
     3361 gggctcgggg gaggggcgcg gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag
     3421 ccgcagccat tgccttttat ggtaatcgtg cgagagggcg cagggacttc ctttgtccca
     3481 aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc
     3541 gaagcggtgc ggcgccggca ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg
     3601 ccgccgtccc cttctccctc tccagcctcg gggctgtccg cggggggacg gctgccttcg
     3661 ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc
     3721 tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat
     3781 tgtgctgtct catcattttg gcaaagaatt cgtcgacgcc gccaccatgg ctatgagcca
     3841 gttcgacatc ctgtgcaaga ccccccccaa ggtgctggtg cggcagttcg tggagagatt
     3901 cgagaggccc agcggcgaga agatcgccag ctgtgccgcc gagctgacct acctgtgctg
     3961 gatgatcacc cacaacggca ccgccatcaa gagggccacc ttcatgagct acaacaccat
     4021 catcagcaac agcctgagct tcgacatcgt gaacaagagc ctgcagttca agtacaagac
     4081 ccagaaggcc accatcctgg aggccagcct gaagaagctg atccccgcct gggagttcac
     4141 catcatccct tacaacggcc agaagcacca gagcgacatc accgacatcg tgtccagcct
     4201 gcagctgcag ttcgagagca gcgaggaggc cgacaagggc aacagccaca gcaagaagat
     4261 gctgaaggcc ctgctgtccg agggcgagag catctgggag atcaccgaga agatcctgaa
     4321 cagcttcgag tacaccagca ggttcaccaa gaccaagacc ctgtaccagt tcctgttcct
     4381 ggccacattc atcaactgcg gcaggttcag cgacatcaag aacgtggacc ccaagagctt
     4441 caagctggtg cagaacaagt acctgggcgt gatcattcag tgcctggtga ccgagaccaa
     4501 gacaagcgtg tccaggcaca tctacttttt cagcgccaga ggcaggatcg accccctggt
     4561 gtacctggac gagttcctga ggaacagcga gcccgtgctg aagagagtga acaggaccgg
     4621 caacagcagc agcaacaagc aggagtacca gctgctgaag gacaacctgg tgcgcagcta
     4681 caacaaggcc ctgaagaaga acgcccccta ccccatcttc gctatcaaga acggccctaa
     4741 gagccacatc ggcaggcacc tgatgaccag ctttctgagc atgaagggcc tgaccgagct
     4801 gacaaacgtg gtgggcaact ggagcgacaa gagggcctcc gccgtggcca ggaccaccta
     4861 cacccaccag atcaccgcca tccccgacca ctacttcgcc ctggtgtcca ggtactacgc
     4921 ctacgacccc atcagcaagg agatgatcgc cctgaaggac gagaccaacc ccatcgagga
     4981 gtggcagcac atcgagcagc tgaagggcag cgccgagggc agcatcagat accccgcctg
     5041 gaacggcatc atcagccagg aggtgctgga ctacctgagc agctacatca acaggcggat
     5101 caccggttgc gtacgcggat cctctgctgg agacatgaga gctgccaacc tttggccaag
     5161 cccgctcatg atcaaacgct ctaagaagaa cagcctggcc ttgtccctga cggccgacca
     5221 gatggtcagt gccttgttgg atgctgagcc ccccatactc tattccgagt atgatcctac
     5281 cagacccttc agtgaagctt cgatgatggg cttactgacc aacctggcag acagggagct
     5341 ggttcacatg atcaactggg cgaagagggt gccaggcttt gtggatttga ccctccatga
     5401 tcaggtccac cttctagaat gtgcctggct agagatcctg atgattggtc tcgtctggcg
     5461 ctccatggag cacccagtga agctactgtt tgctcctaac ttgctcttgg acaggaacca
     5521 gggaaaatgt gtagagggca tggtggagat cttcgacatg ctgctggcta catcatctcg
     5581 gttccgcatg atgaatctgc agggagagga gtttgtgtgc ctcaaatcta ttattttgct
     5641 taattctgga gtgtacacat ttctgtccag caccctgaag tctctggaag agaaggacca
     5701 tatccaccga gtcctggaca agatcacaga cactttgatc cacctgatgg ccaaggcagg
     5761 cctgaccctg cagcagcagc accagcggct ggcccagctc ctcctcatcc tctcccacat
     5821 caggcacatg agtaacaaag gcatggagca tctgtacagc atgaagtgca agaacgtggt
     5881 gcccctctat gacctgctgc tggaggcggc ggacgcccac cgcctacatg cgcccactag
     5941 ccgtggaggg gcatccgtgg aggagacgga ccaaagccac ttggccactg cgggctctac
     6001 ttcatcgcat tccttgcaaa agtattacat cacgggggag gcagagggtt tccctgccac
     6061 agcttgatag acgcgtgaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa
     6121 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     6181 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     6241 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     6301 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     6361 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     6421 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     6481 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctatgttg ccacctggat
     6541 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     6601 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     6661 tcggatctcc ctttgggccg cctccccgca tcgataccga gcgctgctcg agagatctac
     6721 gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca
     6781 gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc
     6841 ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca
     6901 acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg
     6961 ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg
     7021 ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg
     7081 ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct
     7141 tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg
     7201 attttgtagg taaccaggcg cgtgaattca ctcctcaggt gcaggctgcc tatcagaagg
     7261 tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg
     7321 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
     7381 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
     7441 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
     7501 atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga
     7561 aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt
     7621 tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg
     7681 ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc
     7741 tcttatggag atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt
     7801 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     7861 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     7921 cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcg acagtactaa gctttactag
     7981 ggacaggatt ggtgacagaa aagccccatc cttaggcctc ctccttccta gtctcctgat
     8041 attgggtcta acccccacct cctgttaggc agattcctta tctggtgaca cacccccatt
     8101 tcctggagcc atctctctcc ttgccagaac ctctaaggtt tgcttacgat ggagccagag
     8161 aggatcctgg gagggagagc ttggcagggg gtgggaggga agggggggat gcgtgacctg
     8221 cccggttctc agtggccacc ctgcgctacc ctctcccaga acctgagctg ctctgacgcg
     8281 gctgtctggt gcgtttcact gatcctggtg ctgcagcttc cttacacttc ccaagaggag
     8341 aagcagtttg gaaaaacaaa atcagaataa gttggtcctg agttctaact ttggctcttc
     8401 acctttctag tccccaattt atattgttcc tccgtgcgtc agttttacct gtgagataag
     8461 gccagtagcc agccccgtcc tggcagggct gtggtgagga ggggggtgtc cgtgtggaaa
     8521 actccctttg tgagaatggt gcgtcctagg tgttcaccag gtcgtggccg cctctactcc
     8581 ctttctcttt ctccatcctt ctttccttaa agagtcccca gtgctatctg ggacatattc
     8641 ctccgcccag agcagggtcc cgcttcccta aggccctgct ctgggcttct gggtttgagt
     8701 ccttggcaag cccaggagag gcgctcaggc ttccctgtcc cccttcctcg tccaccatct
     8761 catgcccctg gctctcctgc cccttcccta caggggttcc tggctctgct ctaagggcga
     8821 attcgcggcc gctaaattca attcgcccta tagtgagtcg tattacaatt cactggccgt
     8881 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     8941 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     9001 acagttgcgc agcctatacg tacggcagtt taaggtttac acctataaaa gagagagccg
     9061 ttatcgtctg tttgtggatg tacagagtga tattattgac acgccggggc gacggatggt
     9121 gatccccctg gccagtgcac gtctgctgtc agataaagtc tcccgtgaac tttacccggt
     9181 ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc gatatggcca gtgtgccggt
     9241 ctccgttatc ggggaagaag tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc
     9301 cattaacctg atgttctggg gaatataaat gtcaggcatg agattatcaa aaaggatctt
     9361 cacctagatc cttttcacgt agaaagccag tccgcagaaa cggtgctgac cccggatgaa
     9421 tgtcagctac tgggctatct ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc
     9481 ttgcagtggg cttacatggc gatagctaga ctgggcggtt ttatggacag caagcgaacc
     9541 ggaattgcca gctggggcgc cctctggtaa ggttgggaag ccctgcaaag taaactggat
     9601 ggctttcttg ccgccaagga tctgatggcg caggggatca agctctgatc aagagacagg
     9661 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     9721 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     9781 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     9841 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     9901 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     9961 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
    10021 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
    10081 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
    10141 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
    10201 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat gcatggcgat gcctgcttgc
    10261 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
    10321 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
    10381 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
    10441 tcgccttcta tcgccttctt gacgagttct tctgaattat taacgcttac aatttcctga
    10501 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatcagg tggcactttt
    10561 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
    10621 ccgctcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
    10681 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
    10741 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
    10801 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
    10861 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
    10921 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
    10981 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
    11041 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
    11101 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
    11161 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
    11221 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgaatac
    11281 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
    11341 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
    11401 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
    11461 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
    11521 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
    11581 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgacc
    11641 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
    11701 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
    11761 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
    11821 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
    11881 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
    11941 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
    12001 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
    12061 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
    12121 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
    12181 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
    12241 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
    12301 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
    12361 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
    12421 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
    12481 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
    12541 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
    12601 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
    12661 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagctcaga
    12721 attaaccctc actaaaggga ctag
//
