LOCUS       sequence-407654-       18343 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Knock-in vector to insert a motor neuron-specific MACS-sortable 
            genetic reporter into the AAVS1 locus in human pluripotent stem 
            cells. Allows isolation of human iPSC-derived motor neurons..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 18343)
  AUTHORS   Harley P, Kerins C, Gatt A, Neves G, Riccio F, Machado CB, 
            Cheesbrough A, R'Bibo L, Burrone J, Lieberam I
  TITLE     Aberrant axon initial segment plasticity and intrinsic excitability 
            of ALS hiPSC motor neurons.
  JOURNAL   Cell Rep. 2023 Dec 26;42(12):113509. doi: 
            10.1016/j.celrep.2023.113509. Epub 2023 Nov 28.
  PUBMED    38019651
REFERENCE   2  (bases 1 to 18343)
  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 Rep.";
            date: "2023-12-26"; volume: "42(12):113509. doi"; pages: " 
            10.1016/j.celrep.2023.113509. Epub 2023 Nov 28"
COMMENT     Sequence Label: AAVS1-Hb9-CD14
FEATURES             Location/Qualifiers
     source          1..18343
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          8624..8853
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     8836..8855
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     polyA_signal    10089..10313
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(10457..10476)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    10522..10577
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(10576..10595)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(10938..10954)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    10962..10978
                     /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(10986..10992,10993..11010,11011..11016))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11031..11052
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    11126..11962
                     /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(12000..12019)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(12001..12019)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(12026..12043)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(12026..12042)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12035..12057)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             12180..12482
                     /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     12361..12380
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     complement(12885..12904)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     13499..13518
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(13879..14670,14671..14739))
                     /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     14502..14521
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      14863..15451
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     15352..15371
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     15605..15622
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    15739..15760
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(15775..15780,15781..15798,15799..15805)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    15813..15829
                     /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     15837..15853
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        15874..15892
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    15931..16734
                     /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    16741..16766
                     /label=SA
                     /note="splice acceptor site"
     CDS             16790..16843
                     /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             16853..17452
                     /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(16853..16872)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     17349..17369
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    17490..17714
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        17768..17786
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gggacagtgc acttttcctt tcttcatgca agggccaggg ctagaggtcc ctggactcag
       61 tcaagaagtg gagagagctg agggaggcct gggaggccct tattctgtac ccagtgtagc
      121 acacctgcag ccctgattgg gggggggggg cgggggcaat tcacaccctg tattctcacc
      181 aggctaaagt ttcttctatc tggttccttc tctccaaaaa atcagtcaga agggacctcc
      241 cttggctgag agtagaagtg aggtggtcag cgtgctgcag acaagcctgt cctccatctg
      301 cctaaggggt ctgatatcct acaccaagcc ccccctaccc cttctggaga cacataggag
      361 ataagcattt tcaaaaccac catggcactc tcctgttctc ccctgcaccc cttcctccca
      421 gactccatcc tgcctgagga tgatttggtc caacttgaat aatagagtac tctctttcca
      481 catctttcca catcggaaag agtagtccgg gatccagtcc acctgcagcc ctgattaaag
      541 gaaggaagga agactagagg cgcagttcta cctattatac ttattttaag gttcccaggt
      601 cagagatgcc tcatggacat ttctccctaa ggcaggtcct gaccagccct ccccctgaat
      661 ggtgctgatc tgtgacttgt tgcttctatt gaacaaatac agctttgatt tgtttccagc
      721 cagagtggtt agctgatgaa ttgacaaaaa ctaatcagct ttattgggaa acaggtttaa
      781 gggcacggac gtgtcaataa cgctcagcct gaccccctct tccattagct aggcaggctg
      841 attagagtaa gaagaccctc tcgccgattg caccgaaatt gctttcataa actcacaata
      901 ttagcagtcg atgtgagcag ctaatttaga gaagacatcc aggcccgtca catcgatcat
      961 cgggtttatg tatcattatc tcatttacag aatgtcgcag tgtaatacat gtttttggca
     1021 ataggcttgg ccgctcgccg gagccgcgcg ctgttccttt cggccacact gcaggggact
     1081 tcctcaacaa tctacgatgt ctcttcgcgt gaacaaggag gcactttgca cacatatgca
     1141 cacttgcgcc caggctctca gtgcgcgctc agtgcacaca tgcacgtgga cacatgctcc
     1201 gacatactac ctgagctgcc tttccttctc atgcggggat gctggcattg ctctctcctg
     1261 cgagcccctg gtctgaagcc caatgccgga cgctcaccaa ccttggagtc aacctcggag
     1321 agctcaaggt tcggttctgt caccagcata cagagatggt gggtggcggc gcaatcagcc
     1381 accttctctc aaggctttgg tcgcccagtt ggcgggtgtc cagaaatcca caggcctgcg
     1441 aaggtccttc tccgccctcc caggctccag cttgacctgc cctgccctgg gcgggccttt
     1501 ctcgcaacac ttccaggctc agccagatgc gcctggtctt agggagtggg ccacctgttc
     1561 ctttgggtaa ttattttaat gtaaggggtc tacttcaggc ctcctgcagt gacagacggc
     1621 agcggagctg caaagcgctc tcagcctttc agtggaattt cccagatggg ccaaggggag
     1681 gccagggcgg gtcaccaggg gccctttaac gagttcatta gccaaattat cttgcctgca
     1741 gcaaggaccc cagagactgt agggagagta ggggagaagg tgggtctttc tccctccctg
     1801 ctagtctgca agaacagttg tgggaaggaa agagaaagga catcctcccc aaaggggcct
     1861 aggaacacta aaggcaaaga tggcatctct gatgtgactt atgcgccaga ccagcgaggt
     1921 ggtggagtgg tgggaaggaa tcataatgat taggaaacac ggagattcta acaaccaggg
     1981 cactgaccca tattgtggaa caaacctagt gcagaaatca aagtcagagc ccctggaact
     2041 ttattctgtg cttcttaagc aaagaacctg gacatcttgg agccttggtc tgtccgtcca
     2101 tctgtagagt agaaggtagg tgtgtttcct tccacaattt gtgagatcac tgccctaaga
     2161 aaagcaactc tgggaagagc ttctggcatg accctgttgg gtccttcggg cccccttact
     2221 cctgtggtag gaggctttgg caggttgagg tgagcctgat cctagatgcc tgtgctggaa
     2281 aatgaaccca actctcacct gtcgtcgcgc cccccacccc caccccacag ctgtgaggcg
     2341 ctgctctgga ggtagctcag catccccgcc cctcctcctt gtcctcactc tgcctcctgt
     2401 agtgttagct tctgctacct cagcaagtag tgagtgaata gaccaattag ccaggacttc
     2461 ctgccattat ctgctgtaga cttgggcatt agtacgccct gattcaccat aatttcaagt
     2521 cagaacacac taaaaaattc actcaccgag gcattaaaat gttcccagac aacgtctgtg
     2581 tttgtgtaac tgggtttctt cccttcgtgt ttgatgcaca ccagcatcat ataggtcagt
     2641 ggtggagtcc aggctaggca gacaattttt gtcgcttttt gtttgttttt atatttttaa
     2701 aacatgaatt gcattgtttc ccggtggttg taaactttgt ttctggggct tcagtattgc
     2761 cagtgtaatc ttcttctgct tgatcagggg caaacaagtg acttagaaag ttgtgggagt
     2821 gccctaagaa cagtagccag agcctttaca gagccaggct ttgtgtccta aacgtacatc
     2881 gccccagcta tgggaagctg ggttatcacc ttcaaagcag accctttctg tggatgactt
     2941 accaagccct gcaggtgggt agtggaggtg gggagagggc tgcacagcct gcttggagct
     3001 tgcttctgtt ctttgtcctg cccatgatat gaagctccta tacaatgaat ttgcagtaag
     3061 atcttggagc aaaagtacag gtttcctgag aagtctttcg tttcttctga gggtgctgtt
     3121 ttacttacac acacacacac acacacacac acacacacac acacacacac accctagctg
     3181 attaaaagaa ctgaggtgac ctctcttagc cttgtttctc tttatgattc atacctatgg
     3241 cctctaggat tgtaagagaa attgaaaatg tctttggctt cctgggtcag aaagcatcca
     3301 gcgcagggct gagctcagtc tcttctctat gttctaagta cacagcaaaa caaacaaaaa
     3361 cccgaaacct tgagtaaaat tttcaaatat agaatatatt tttgtgctgg agtttttcaa
     3421 atttatatct tactgtcttt ctccctgtac ccctacaacc caggtttggg ctggacttat
     3481 ttatttattt acaaggagga atggggccag tggagaaggg cagccagtga agatggcctg
     3541 agggtggacc aggcagtagg gagggcagag taagctttct aaaaagggag gaaggaaatc
     3601 caagtgtgct gggagaggta gagaaggggg gggggctggc aaagaaaagg actggcttgt
     3661 agtgtccttg gcaagagcag ttcacctcag gttggaacac tgggtcttca ggtcacctgt
     3721 cttctgagcc acctttgcaa accacccctt taaagctgaa ctggggatgg aggtctgaga
     3781 aaagggtggt agatcaaagg atggtaataa aggagctgat ggtacttttg ggctctgaag
     3841 gggctgtaat gagtgtgtgt gtgtgggggg ggactcatca gacatccctt ctcctgatct
     3901 cacctccctg tgaccccagg ctccccattg gattctgtgt gtcctcaggc tggggtgggt
     3961 ggacgggtta tgtccccaga gacattccaa gcactgattt cagtctgact tgaaggaagt
     4021 tggtctatga ccttccaggt ccagcattca cacttcctgc ttgctttctg tcctcctgag
     4081 tacttattca gcctctgaaa aataccacca tcaccacccc tgtgggcttg tgtcctggac
     4141 ctcactggcc tcactacatc ctccaggctc attttcctga agggaagcct taaaatgttt
     4201 ttttttttaa agatttattt atttattata tgtaagtaca ttgtagctgt cttcagacac
     4261 accagaagag ggagtcagat ctcgttacag atggttgtga gccaccatgt ggttgctggg
     4321 atttgaactc tggaccttcg gaagagcagt agggtgctct aacccactga gccatctcac
     4381 cagccccgcc tcaaaatgtt gatgtgagtt aactctcatg gagttcaaaa gggctgagaa
     4441 gtcagtcttc atgaaccttt ctccatgagg ctgctgtggg cagggggttg cttgttttaa
     4501 gttttggaaa cagccagaca aatggcaggt gtgtggaggg agatgcaggg aatgctgcct
     4561 tccatccagt ctcctagcca tacctccagg ctcttggcct ttttgttaat ggctattata
     4621 tattgtacct gctgcattcc acagaaaggg atgtgtttgt aggtgatcag ttatagaatc
     4681 aatgctgacc tgacctgtga atatgggtgc atgtaggttt tgttctgagt tcttgtataa
     4741 tgtcaggggt aaaacttgta gattataaca tctttgtaag ttttcttttt atgtttgcat
     4801 tgtgtgtatg tttgtgtcta tttatgggaa tgcacgtacc caaagaggcc ataagaggga
     4861 atccgatttc ctgtagctgg agttggaggt ggttgtgagt gtgagtggtg ggaacccaac
     4921 ttgggtcctc ctcaagagct gcactcactc ttaagcactg aactgtggaa ccactatact
     4981 ccccacccca acatcttatt tccttgtggt tatttgtata aatcaacaca cctttctcag
     5041 taatgagagg aggctcaatc acaagttata ttcagtaact taatgcaaga cccaaaagag
     5101 ggatcctggc tagacgcaga gaaaaaaatg actgggagcc agatgattaa atgtacaccg
     5161 ggccttaaat tccagcctct tttgggtagg agaggtcttc ctctctgccc tccaagagcc
     5221 tcatttggcc cctggggctt ctggagaagg gagctacctt gaggcacaac aatgggaatg
     5281 ctgtgctcac ttcggcagca cctatactaa aactgaaccg atacagagaa gattagcatg
     5341 gcccctgcgc aaggatgaca tgcaaatttg tgaagcattc cttatttttt tttgccagga
     5401 ccaggaatgg gaacaggtgg gttggggagc agagggaggg gagaggggat aggggatttg
     5461 gggaggggaa actaggaaag gggataacat ttgaaatgta aataaagaaa atatctaata
     5521 aaaaaaaaaa aaagaatagg catgcttctg ggaccagacc tttgatctct caagggtacc
     5581 gtagtaaacc cccagctcag tcccaggctg ccccagagcc agctgggctg ggtcaataga
     5641 cgctgggaac aggaagagac gcagcatgct cactccactc gtaaaacaca caaagatccc
     5701 agttccagac gactcccagt ctctctccca actcccacct ctcaggatac aagttcccgc
     5761 tcaaaccgaa gcagaatata tgatcatatt caagcccttt gacttcagag tctttttggc
     5821 ccaaccccca ctttgtcaaa caaagaaata tatccatcaa gtctgatatg caaatttccc
     5881 cttcgctgaa ggggctcttg atacaacctc actcagccca caggtggtgg ggactggtgt
     5941 gggttagcct ggtggatggt caaacaccag acgaagtggg aagggtttga aagggattta
     6001 gaagcttttg gatcacatcc cctacacaac aaacaggaaa tgaattcctc ttctcggatg
     6061 actttactaa ggaaacctgt gcctaaactc agtagaccta atactgtatc atcaccaaaa
     6121 acaaccctgg ttgtgtagtt tagagatgat tcgggtccat gaggtccacc ccaggaaaat
     6181 ctcatctgca gccacccttt ttttcctcct tcttctcctt tgtttgtttt gttgttgttt
     6241 ttttcaagac agggtatctc tgtgtagcat ttttggttgc tgtcctggaa ctggctctgt
     6301 agactaggct ggcctagaac tcacagaccc cacctgcctc tgcctcctga gtgctgggat
     6361 tagagacttg cctcaccatg gcttaaacac aagtttctgt aaatgcaatg gggtcaagac
     6421 tgaatcctgg ggatgggggt agggcatccc agcatcctta cttgccttaa ttgggcttcc
     6481 aagataggag aactggcagg gaaccactca tggaacacaa gtgttccttt aaccctctaa
     6541 aaggcagagc tgcattttca tcaccccaaa gagacagcac ctaaagacaa gagctatgtg
     6601 cttgtactat tttcagacta gctggaggcc tttgcaaagc tgtgcacaac ttactccaag
     6661 atgttccccg ctgggttcca gattttccag ccaggaccac cctgtcataa taaaaactcc
     6721 actcagccga gcatggtggc gcacaccttt aatcccagca cttgggaggc agaggcagga
     6781 ggatttctga gttcgaggcc agcctggtct acagagtgag ttccaagaca gccagggcta
     6841 gacagagaaa ctctgtctcc aaaatcaaaa acaaaaacaa aacaaaaaca acaaaaacaa
     6901 aaaacctact cactcatccc aacagaatca ctgcgttaaa tctctcctaa acaaaacttg
     6961 ttgacaggga aaatatagtc cttacttata tttatcgtta ccccccaccc ccaatccctc
     7021 ttgaaacagc tttgcctgct acttttaaat tcacccaaca aggattgtcc tagcctcctg
     7081 gccttccctg ggccatttcc gatcttttat taacctctca cctggttggg aagccgcgat
     7141 ccagagacct ggaactccct cccactttcc cctcgtgttt ggggaagata cccgagaatg
     7201 gagctagcga gaaagcttga tttctcaaac tacttattga ttcgttacat taaaaaaaaa
     7261 aagcccccgg aagcgtgttt acagtaaata tttgatatgt gcgcaggctg aggtctggat
     7321 tcacacttta atacggtctc tggaagccct ttcttgattt tcctgaactc ggccggcagg
     7381 aggaggcgag agcctgatag tggctccctt agcctgcgaa ctggctcagt ccgagggagc
     7441 agtcctcggc caattcccgc aaagcctttt tcctacaccc ccacaaggca aggctctaat
     7501 agaacttggt tctcacctag gtccggctca gtggcacctg tgcaaacgac cttcaagtca
     7561 atgtgcatct ctgagttgga aaactgctct ctctccactc tgactgaaga tttgtaggct
     7621 tcaacctcta ataccgtatt tatacacacc gaaaccctac aaacaaacat ttgcggcgca
     7681 tgatactttt gggggagggg cagataagaa ggttcgacct gctgagggtt aatgacaggg
     7741 acgcaaacca tcttgaagca atcaacactc aggaacgcga gagattttgg gaaaccgaac
     7801 agtcttcagt gtactaagct cagggtcccg ttttgggtac ctagtggact tcaggggtga
     7861 ggacccacct agttcctcag tgcagaacat tccacaactc cctcaaggga gcgcgagtag
     7921 ctggaggaag ataaggcgtc cgcctctgga ggaacaaggt gtcaaaggga ctcggggggc
     7981 actcactggg acccagggtt actggcgctc accactctcc cagcagcaag gaagcagtaa
     8041 gcttgtgttc agcctccaaa ctctccaacc tgcctcctcc ggccacctac gctgttcacc
     8101 accggctgct taggacctgg gggcgcaggg gcggggcgtc aggctaattg gaccgtgaca
     8161 ggtgagcccc tcggccaatc gccgggcaga cagccctcct cgggcccccg ctgagcgcgc
     8221 caatcccaga ccgccctgag cttcaagtgt gcggaggggc gcgggtcccc accaaggcac
     8281 cagtggcctc gatgcggtcg cttgagctct tgagagcaga gagccggtct ccccaacaca
     8341 aactcgcagg agcgctccgg tggaagttca taccagtgag tggagcgcgg tccccacgag
     8401 gaccgcaaag aggaggcgcc tggacgatgg tcgctctcag ctgagtttcc ggctgcgact
     8461 ttattggcaa aaaatcgcag taacaatacc ggccccaagg ctggccaccg ccacgctcag
     8521 tctccgtaag cccgcacacg actgcatcgc ccacccttgg accctgattg tcagctccaa
     8581 tcgagccggc gcgcctcgct gtctgcgagg gccagctgtt ggggtgagta ctccctctca
     8641 aaagcgggca tgacttctgc gctaagattg tcagtttcca aaaacgagga ggatttgata
     8701 ttcacctggc ccgcggtgat gcctttgagg gtggccgcgt ccatctggtc agaaaagaca
     8761 atctttttgt tgtcaagctt gaggtgtggc aggcttgaga tctggccata cacttgagtg
     8821 acaatgacat ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcagt
     8881 ccaagcggat cacccaccgg tccaccatgg agcgcgcgtc ctgcttgttg ctgctgctgc
     8941 tgccgctggt gcacgtctct gcgaccacgc cagaaccttg tgagctggac gatgaagatt
     9001 tccgctgcgt ctgcaacttc tccgaacctc agcccgactg gtccgaagcc ttccagtgtg
     9061 tgtctgcagt agaggtggag atccatgccg gcggtctcaa cctagagccg tttctaaagc
     9121 gcgtcgatgc ggacgccgac ccgcggcagt atgctgacac ggtcaaggct ctccgcgtgc
     9181 ggcggctcac agtgggagcc gcacaggttc ctgctcagct actggtaggc gccctgcgtg
     9241 tgctagcgta ctcccgcctc aaggaactga cgctcgagga cctaaagata accggcacca
     9301 tgcctccgct gcctctggaa gccacaggac ttgcactttc cagcttgcgc ctacgcaacg
     9361 tgtcgtgggc gacagggcgt tcttggctcg ccgagctgca gcagtggctc aagccaggcc
     9421 tcaaggtact gagcattgcc caagcacact cgcctgcctt ttcctgcgaa caggttcgcg
     9481 ccttcccggc ccttaccagc ctagacctgt ctgacaatcc tggactgggc gaacgcggac
     9541 tgatggcggc tctctgtccc cacaagttcc cggccatcca gaatctagcg ctgcgcaaca
     9601 caggaatgga gacgcccaca ggcgtgtgcg ccgcactggc ggcggcaggt gtgcagcccc
     9661 acagcctaga cctcagccac aactcgctgc gcgccaccgt aaaccctagc gctccgagat
     9721 gcatgtggtc cagcgccctg aactccctca atctgtcgtt cgctgggctg gaacaggtgc
     9781 ctaaaggact gccagccaag ctcagagtgc tcgatctcag ctgcaacaga ctgaacaggg
     9841 cgccgcagcc tgacgagctg cccgaggtgg ataacctgac actggacggg aatcccttcc
     9901 tggtccctgg aactgccctc ccccacgagg gctcaatgaa ctccggcgtg gtcccagcct
     9961 gtgcacgttc gaccctgtcg gtgggggtgt cgggaaccct ggtgctgctc caaggggccc
    10021 ggggctttgc ctaaactagt ggatcccatg ggggcccgct agccctagag ctcgctgatc
    10081 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
    10141 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
    10201 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
    10261 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
    10321 ggcggaaaga accagctggg gctcgagtct agtttaatta agatatcacg cgtgaattca
    10381 ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg tgtggccaat gccctggctc
    10441 acaaatacca ctgagatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc
    10501 ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga
    10561 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga
    10621 atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag
    10681 gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc
    10741 atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt
    10801 ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct
    10861 gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac ctgcagccca
    10921 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
    10981 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
    11041 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
    11101 cagcggatcg acagtactaa gctttactag ggacaggatt ggtgacagaa aagccccatc
    11161 cttaggcctc ctccttccta gtctcctgat attgggtcta acccccacct cctgttaggc
    11221 agattcctta tctggtgaca cacccccatt tcctggagcc atctctctcc ttgccagaac
    11281 ctctaaggtt tgcttacgat ggagccagag aggatcctgg gagggagagc ttggcagggg
    11341 gtgggaggga agggggggat gcgtgacctg cccggttctc agtggccacc ctgcgctacc
    11401 ctctcccaga acctgagctg ctctgacgcg gctgtctggt gcgtttcact gatcctggtg
    11461 ctgcagcttc cttacacttc ccaagaggag aagcagtttg gaaaaacaaa atcagaataa
    11521 gttggtcctg agttctaact ttggctcttc acctttctag tccccaattt atattgttcc
    11581 tccgtgcgtc agttttacct gtgagataag gccagtagcc agccccgtcc tggcagggct
    11641 gtggtgagga ggggggtgtc cgtgtggaaa actccctttg tgagaatggt gcgtcctagg
    11701 tgttcaccag gtcgtggccg cctctactcc ctttctcttt ctccatcctt ctttccttaa
    11761 agagtcccca gtgctatctg ggacatattc ctccgcccag agcagggtcc cgcttcccta
    11821 aggccctgct ctgggcttct gggtttgagt ccttggcaag cccaggagag gcgctcaggc
    11881 ttccctgtcc cccttcctcg tccaccatct catgcccctg gctctcctgc cccttcccta
    11941 caggggttcc tggctctgct ctaagggcga attcgcggcc ggccgctaaa ttcaattcgc
    12001 cctatagtga gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
    12061 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
    12121 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagccta tacgtacggc
    12181 agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga
    12241 gtgatattat tgacacgccg gggcgacgga tggtgatccc cctggccagt gcacgtctgc
    12301 tgtcagataa agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc
    12361 gcatgatgac caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg
    12421 atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat
    12481 aaatgtcagg catgagatta tcaaaaagga tcttcaccta gatccttttc acgtagaaag
    12541 ccagtccgca gaaacggtgc tgaccccgga tgaatgtcag ctactgggct atctggacaa
    12601 gggaaaacgc aagcgcaaag agaaagcagg tagcttgcag tgggcttaca tggcgatagc
    12661 tagactgggc ggttttatgg acagcaagcg aaccggaatt gccagctggg gcgccctctg
    12721 gtaaggttgg gaagccctgc aaagtaaact ggatggcttt cttgccgcca aggatctgat
    12781 ggcgcagggg atcaagctct gatcaagaga caggatgagg atcgtttcgc atgattgaac
    12841 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
    12901 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
    12961 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
    13021 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
    13081 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
    13141 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
    13201 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
    13261 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
    13321 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
    13381 tcgtcgtgac ccatgcatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
    13441 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
    13501 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
    13561 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
    13621 ttcttctgaa ttattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg
    13681 tgcggtattt cacaccgcat caggtggcac ttttcgggga aatgtgcgcg gaacccctat
    13741 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagattat caaaaaggat
    13801 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
    13861 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
    13921 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
    13981 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
    14041 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
    14101 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
    14161 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
    14221 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
    14281 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
    14341 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
    14401 atccgtaaga tgcttttctg tgactggtga atactcaacc aagtcattct gagaatagtg
    14461 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
    14521 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
    14581 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
    14641 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
    14701 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
    14761 ttgaagcatt tatcagggtt attgtctcat gaccaaaatc ccttaacgtg agttttcgtt
    14821 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
    14881 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
    14941 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
    15001 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
    15061 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
    15121 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
    15181 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
    15241 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
    15301 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
    15361 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
    15421 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
    15481 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
    15541 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
    15601 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
    15661 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
    15721 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
    15781 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
    15841 aaacagctat gaccatgatt acgccaagct cagaattaac cctcactaaa gggactagct
    15901 agtcctgcag gtttaaacga attcgccctt tgctttctct gaccagcatt ctctcccctg
    15961 ggcctgtgcc gctttctgtc tgcagcttgt ggcctgggtc acctctacgg ctggcccaga
    16021 tccttccctg ccgcctcctt caggttccgt cttcctccac tccctcttcc ccttgctctc
    16081 tgctgtgttg ctgcccaagg atgctctttc cggagcactt ccttctcggc gctgcaccac
    16141 gtgatgtcct ctgagcggat cctccccgtg tctgggtcct ctccgggcat ctctcctccc
    16201 tcacccaacc ccatgccgtc ttcactcgct gggttccctt ttccttctcc ttctggggcc
    16261 tgtgccatct ctcgtttctt aggatggcct tctccgacgg atgtctccct tgcgtcccgc
    16321 ctccccttct tgtaggcctg catcatcacc gtttttctgg acaaccccaa agtaccccgt
    16381 ctccctggct ttagccacct ctccatcctc ttgctttctt tgcctggaca ccccgttctc
    16441 ctgtggattc gggtcacctc tcactccttt catttgggca gctcccctac cccccttacc
    16501 tctctagtct gtgctagctc ttccagcccc ctgtcatggc atcttccagg ggtccgagag
    16561 ctcagctagt cttcttcctc caacccgggc ccctatgtcc acttcaggac agcatgtttg
    16621 ctgcctccag ggatcctgtg tccccgagct gggaccacct tatattccca gggccggtta
    16681 atgtggctct ggttctgggt acttttatct gtcccctcca ccccacagtg gggcaagctt
    16741 ctgacctctt ctcttcctcc cacagggcct cgagagatct ggcagcggag agggcagagg
    16801 aagtcttcta acatgcggtg acgtggagga gaatcccggc cctaggctcg agatgaccga
    16861 gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct
    16921 cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat
    16981 cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa
    17041 ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga
    17101 agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct
    17161 ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg
    17221 gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt
    17281 cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc
    17341 cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga
    17401 ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gatctagagg
    17461 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
    17521 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
    17581 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
    17641 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
    17701 ggtgggctct atgggtctcg acattgatta ttgactagca ctagtgggcc catgcggccg
    17761 cgagctcaat taaccctcac taaagggagt cgactcgatc ctgtctccct ggccaggagt
    17821 ctggggtccg cagatatatt acaacaaaat tactaattgg aaaatgaaat tgtcaataaa
    17881 gcacacccca cgcgtgtact atatctttaa atattatgat gtcttattgg tcccatttcc
    17941 ttacaataat ttattgcaat aaaacagaca ctgcaggtcc ccttaaattg tcttttcata
    18001 atgaataaat ttaagcaggc taattaatat ataaactagc tcaatttgtc aagttgattt
    18061 gtattttagt taattgtgaa agtaattacc acatggtcaa attaacagct ttctggaaat
    18121 gaccaagcct gaggttttat ttccttcctg ggtgaagaaa attcattttt ccaagctctt
    18181 gatgtgatga ataaaagtca taaatctggg tgattggtgc aggcagagtc taaatggctt
    18241 catatttcat tttaggttta atagaaatat tcatgctctg ttttaatgaa attaaattga
    18301 agggggatgg ggctctggag gcggccctgg ctgggggctt aaa
//
