LOCUS       sequence-418877-        9757 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  donor vector for AAVS1 targeting (puromycin selection) and 
            constitutive GFP expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9757)
  AUTHORS   Oceguera-Yanez F, Kim SI, Matsumoto T, Tan GW, Xiang L, Hatani T, 
            Kondo T, Ikeya M, Yoshida Y, Inoue H, Woltjen K
  TITLE     Engineering the AAVS1 locus for consistent and scalable transgene 
            expression in human iPSCs and their differentiated derivatives.
  JOURNAL   Methods. 2015 Dec 18. pii: S1046-2023(15)30181-X. doi: 
            10.1016/j.ymeth.2015.12.012.
  PUBMED    26707206
REFERENCE   2  (bases 1 to 9757)
  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: "Methods. 
            2015 Dec 18. pii: S1046-2023(15)30181-X. doi: 
            10.1016/j.ymeth.2015.12.012."
COMMENT     Sequence Label: pAAVS1-P-CAG-GFP
FEATURES             Location/Qualifiers
     source          1..9757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     89..106
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    223..244
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(259..264,265..282,283..289)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    297..313
                     /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     302..324
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     321..337
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     321..337
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        358..376
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    411..1214
                     /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    1221..1246
                     /label=SA
                     /note="splice acceptor site"
     CDS             1270..1323
                     /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             1333..1932
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1333..1352)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1829..1849
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(1964..1981)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1970..2194
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        2198..2577
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          3001..4017
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3941..3963
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     4025..4044
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      4072..4081
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4078..4080,4081..4083,4084..4797)
                     /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             join(4078..4080,4081..4083,4084..4794)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4123..4144)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4384..4403)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4731..4752
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4881..4900)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4946..5001
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5000..5019)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     misc_feature    5375..6211
                     /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(6245..6264)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6246..6264)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6271..6288)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6271..6287)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6280..6302)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             6425..6727
                     /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     6606..6625
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             7076..7870
                     /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             7076..7870
                     /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(7130..7149)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7740..7759
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(8120..8911,8912..8980))
                     /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     8743..8762
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9104..9692
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9593..9612
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
       61 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
      121 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
      181 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
      241 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
      301 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agctcagaat
      361 taaccctcac taaagggact agtcctgcag gtttaaacga attcgccctt tgctttctct
      421 gaccagcatt ctctcccctg ggcctgtgcc gctttctgtc tgcagcttgt ggcctgggtc
      481 acctctacgg ctggcccaga tccttccctg ccgcctcctt caggttccgt cttcctccac
      541 tccctcttcc ccttgctctc tgctgtgttg ctgcccaagg atgctctttc cggagcactt
      601 ccttctcggc gctgcaccac gtgatgtcct ctgagcggat cctccccgtg tctgggtcct
      661 ctccgggcat ctctcctccc tcacccaacc ccatgccgtc ttcactcgct gggttccctt
      721 ttccttctcc ttctggggcc tgtgccatct ctcgtttctt aggatggcct tctccgacgg
      781 atgtctccct tgcgtcccgc ctccccttct tgtaggcctg catcatcacc gtttttctgg
      841 acaaccccaa agtaccccgt ctccctggct ttagccacct ctccatcctc ttgctttctt
      901 tgcctggaca ccccgttctc ctgtggattc gggtcacctc tcactccttt catttgggca
      961 gctcccctac cccccttacc tctctagtct gtgctagctc ttccagcccc ctgtcatggc
     1021 atcttccagg ggtccgagag ctcagctagt cttcttcctc caacccgggc ccctatgtcc
     1081 acttcaggac agcatgtttg ctgcctccag ggatcctgtg tccccgagct gggaccacct
     1141 tatattccca gggccggtta atgtggctct ggttctgggt acttttatct gtcccctcca
     1201 ccccacagtg gggcaagctt ctgacctctt ctcttcctcc cacagggcct cgagagatct
     1261 ggcagcggag agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc
     1321 cctaggctcg agatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     1381 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     1441 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     1501 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     1561 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     1621 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     1681 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     1741 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     1801 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     1861 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     1921 cccggtgcct gatctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     1981 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     2041 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     2101 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     2161 agcaggcatg ctggggatgc ggtgggctct atgggtcgac attgattatt gactagttat
     2221 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     2281 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     2341 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     2401 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     2461 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     2521 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc
     2581 gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt
     2641 ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg gggggggggg
     2701 gcgggccagg gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg
     2761 ggggcggccg ccgacggcgg gccgccccgg ggagttcctt tttttggggg ggggggggcg
     2821 gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg gggggggggg
     2881 ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
     2941 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
     3001 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
     3061 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
     3121 ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa
     3181 agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc
     3241 gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc
     3301 tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg
     3361 ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
     3421 tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac
     3481 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt
     3541 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg
     3601 gggccgcctc gggccggggg gggctcgggg gaggggcgcg gcggcccccg gagcgccggc
     3661 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
     3721 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc
     3781 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag
     3841 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg
     3901 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
     3961 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
     4021 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt cgccaccatg
     4081 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     4141 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     4201 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     4261 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     4321 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     4381 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     4441 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     4501 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     4561 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     4621 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     4681 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     4741 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaagaa
     4801 ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc caatgccctg
     4861 gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga catcatgaag
     4921 ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt
     4981 tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat
     5041 cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac
     5101 aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta
     5161 ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt
     5221 ttttctttaa catccctaaa attttcctta catgttttac tagccagatt tttcctcctc
     5281 tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct cgacctgcag
     5341 cccaagcttg gatccctcga cagtactaag ctttactagg gacaggattg gtgacagaaa
     5401 agccccatcc ttaggcctcc tccttcctag tctcctgata ttgggtctaa cccccacctc
     5461 ctgttaggca gattccttat ctggtgacac acccccattt cctggagcca tctctctcct
     5521 tgccagaacc tctaaggttt gcttacgatg gagccagaga ggatcctggg agggagagct
     5581 tggcaggggg tgggagggaa gggggggatg cgtgacctgc ccggttctca gtggccaccc
     5641 tgcgctaccc tctcccagaa cctgagctgc tctgacgcgg ctgtctggtg cgtttcactg
     5701 atcctggtgc tgcagcttcc ttacacttcc caagaggaga agcagtttgg aaaaacaaaa
     5761 tcagaataag ttggtcctga gttctaactt tggctcttca cctttctagt ccccaattta
     5821 tattgttcct ccgtgcgtca gttttacctg tgagataagg ccagtagcca gccccgtcct
     5881 ggcagggctg tggtgaggag gggggtgtcc gtgtggaaaa ctccctttgt gagaatggtg
     5941 cgtcctaggt gttcaccagg tcgtggccgc ctctactccc tttctctttc tccatccttc
     6001 tttccttaaa gagtccccag tgctatctgg gacatattcc tccgcccaga gcagggtccc
     6061 gcttccctaa ggccctgctc tgggcttctg ggtttgagtc cttggcaagc ccaggagagg
     6121 cgctcaggct tccctgtccc ccttcctcgt ccaccatctc atgcccctgg ctctcctgcc
     6181 ccttccctac aggggttcct ggctctgctc taagggcgaa ttcgcggccg ctaaattcaa
     6241 ttcgccctat agtgagtcgt attacaattc actggccgtc gttttacaac gtcgtgactg
     6301 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     6361 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctatacgt
     6421 acggcagttt aaggtttaca cctataaaag agagagccgt tatcgtctgt ttgtggatgt
     6481 acagagtgat attattgaca cgccggggcg acggatggtg atccccctgg ccagtgcacg
     6541 tctgctgtca gataaagtct cccgtgaact ttacccggtg gtgcatatcg gggatgaaag
     6601 ctggcgcatg atgaccaccg atatggccag tgtgccggtc tccgttatcg gggaagaagt
     6661 ggctgatctc agccaccgcg aaaatgacat caaaaacgcc attaacctga tgttctgggg
     6721 aatataaatg tcaggcatga gattatcaaa aaggatcttc acctagatcc ttttcacgta
     6781 gaaagccagt ccgcagaaac ggtgctgacc ccggatgaat gtcagctact gggctatctg
     6841 gacaagggaa aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc ttacatggcg
     6901 atagctagac tgggcggttt tatggacagc aagcgaaccg gaattgccag ctggggcgcc
     6961 ctctggtaag gttgggaagc cctgcaaagt aaactggatg gctttcttgc cgccaaggat
     7021 ctgatggcgc aggggatcaa gctctgatca agagacagga tgaggatcgt ttcgcatgat
     7081 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     7141 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     7201 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     7261 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     7321 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     7381 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     7441 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     7501 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     7561 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     7621 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     7681 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     7741 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     7801 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     7861 gttcttctga attattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct
     7921 gtgcggtatt tcacaccgca tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     7981 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagatta tcaaaaagga
     8041 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     8101 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     8161 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     8221 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     8281 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     8341 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     8401 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     8461 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     8521 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     8581 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     8641 catccgtaag atgcttttct gtgactggtg aatactcaac caagtcattc tgagaatagt
     8701 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     8761 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     8821 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     8881 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     8941 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     9001 attgaagcat ttatcagggt tattgtctca tgaccaaaat cccttaacgt gagttttcgt
     9061 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     9121 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     9181 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     9241 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     9301 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     9361 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     9421 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     9481 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     9541 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     9601 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     9661 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     9721 tcctggcctt ttgctggcct tttgctcaca tgttctt
//
