LOCUS       sequence-200136-       12408 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PiggyBac transposon system construct for expression of 10 gRNAs 
            targeting OCT4, MYC, KLF4, SOX2 and LIN28A promoters and 5 gRNAs 
            targeting EEA-motif. Includes PGK-puro selection cassette..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12408)
  AUTHORS   Weltner J, Balboa D, Katayama S, Bespalov M, Krjutskov K, Jouhilahti
            EM, Trokovic R, Kere J, Otonkoski T
  TITLE     Human pluripotent reprogramming with CRISPR activators.
  JOURNAL   Nat Commun. 2018 Jul 6;9(1):2643. doi: 10.1038/s41467-018-05067-x.
  PUBMED    29980666
REFERENCE   2  (bases 1 to 12408)
  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: "Nat 
            Commun."; date: "2018-07-6"; volume: "9(1):2643. doi"; pages: " 
            10.1038/s41467-018-05067-x"
COMMENT     Sequence Label: PB-EEA-5g-OSK2M2L1-PGK-Puro
FEATURES             Location/Qualifiers
     source          1..12408
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    118..151
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     494..510
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        511..759
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        780..855
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     887..903
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        904..1152
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        1174..1249
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(1261..1277)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1281..1297
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1298..1546
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        1568..1643
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     1675..1691
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1692..1940
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        1962..2037
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     2069..2085
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2086..2334
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        2356..2431
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    2589..2622
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     2965..2981
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2982..3230
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        3252..3327
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     3359..3375
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3376..3624
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        3646..3721
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(3733..3749)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3753..3769
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3770..4018
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        4040..4115
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     4147..4163
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4164..4412
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        4434..4509
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     4541..4557
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4558..4806
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        4828..4903
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    5061..5094
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5155..5654
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     5587..5606
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             5694..6293
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APHNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5694..5713)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     complement(6374..6391)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6380..6604
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6651..6667)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6675..6691
                     /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(6699..6705,6706..6723,6724..6729))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6744..6765
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    complement(6824..6948)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     repeat_region   7238..7272
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(7301..7320)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7302..7320)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7330..7346)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7339..7361)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7488..7943
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7575..7594)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7785..7806
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7969..8073
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8074..8142,8143..8934)
                     /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     complement(8292..8311)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9105..9693
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9594..9613
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    9981..10002
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10017..10022,10023..10040,10041..10047)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10055..10071
                     /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     10079..10095
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10114..10134
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        10116..10134
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   10185..10247
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     10460..10476
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        10477..10725
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        10741..10816
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     10848..10864
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        10865..11113
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        11129..11204
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(11216..11232)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     11236..11252
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        11253..11501
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        11517..11592
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     11624..11640
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        11641..11889
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        11905..11980
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     12012..12028
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        12029..12277
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     misc_RNA        12293..12368
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
       61 ccgggcgacc aaaggtcgcc cgacgcccaa ttccgatcat attcaataac ccttaatata
      121 acttcgtata atgtatgcta tacgaagtta ttaggtctga agaggagttt acgtccagcc
      181 aagcttagga tctcgacctc gaaattctac cgggtagggg aggcgctttt cccaaggcag
      241 tctggagcat gcgctttagc agccccgctg ggcacttggc gctacacaag tggcctctgg
      301 cctcgcacac attccacatc caccggtagg cgccaaccgg ctccgttctt tggtggcccc
      361 ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg
      421 tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta gaggatctcg agctcaagct
      481 tcgaattccc cctgtaaaac gacggccagt gagggcctat ttcccatgat tccttcatat
      541 ttgcatatac gatacaaggc tgttagagag ataattggaa ttaatttgac tgtaaacaca
      601 aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt
      661 ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat
      721 ttcttggctt tatatatctt gtggaaagga cgaaacaccg gggggagaaa ctgaggcgag
      781 ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg
      841 gcaccgagtc ggtgcttttt ttcatggtca tagctgtttc ctcatggtaa aacgacggcc
      901 agtgagggcc tatttcccat gattccttca tatttgcata tacgatacaa ggctgttaga
      961 gagataattg gaattaattt gactgtaaac acaaagatat tagtacaaaa tacgtgacgt
     1021 agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat
     1081 catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat cttgtggaaa
     1141 ggacgaaaca ccgcaaaacc cggcagcgag gctgttttag agctagaaat agcaagttaa
     1201 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct ttttttcatg
     1261 gtcatagctg tttcctggac gtaaaacgac ggccagtgag ggcctatttc ccatgattcc
     1321 ttcatatttg catatacgat acaaggctgt tagagagata attggaatta atttgactgt
     1381 aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt
     1441 tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta
     1501 tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccggtg gctggcaggc
     1561 tggctctgtt ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg
     1621 aaaaagtggc accgagtcgg tgcttttttt catggtcata gctgtttcct ccaggtaaaa
     1681 cgacggccag tgagggccta tttcccatga ttccttcata tttgcatata cgatacaagg
     1741 ctgttagaga gataattgga attaatttga ctgtaaacac aaagatatta gtacaaaata
     1801 cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa
     1861 tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct ttatatatct
     1921 tgtggaaagg acgaaacacc gggcacagtg ccagaggtct ggttttagag ctagaaatag
     1981 caagttaaaa taaggctagt ccgttatcaa cttgaaaaag tggcaccgag tcggtgcttt
     2041 ttttcatggt catagctgtt tccttgttgt aaaacgacgg ccagtgaggg cctatttccc
     2101 atgattcctt catatttgca tatacgatac aaggctgtta gagagataat tggaattaat
     2161 ttgactgtaa acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt
     2221 gggtagtttg cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact
     2281 tgaaagtatt tcgatttctt ggctttatat atcttgtgga aaggacgaaa caccgtctgt
     2341 gggggacctg cactggtttt agagctagaa atagcaagtt aaaataaggc tagtccgtta
     2401 tcaacttgaa aaagtggcac cgagtcggtg ctttttttca tggtcatagc tgtttcctaa
     2461 cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     2521 gctcactgag gccgggcgac caaaggtcgc ccgacgccca attccgatca tattcaataa
     2581 cccttaatat aacttcgtat aatgtatgct atacgaagtt attaggtctg aagaggagtt
     2641 tacgtccagc caagcttagg atctcgacct cgaaattcta ccgggtaggg gaggcgcttt
     2701 tcccaaggca gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa
     2761 gtggcctctg gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct
     2821 ttggtggccc cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc
     2881 cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agaggatctc
     2941 gagctcaagc ttcgaattcc ccctgtaaaa cgacggccag tgagggccta tttcccatga
     3001 ttccttcata tttgcatata cgatacaagg ctgttagaga gataattgga attaatttga
     3061 ctgtaaacac aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt
     3121 agtttgcagt tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa
     3181 agtatttcga tttcttggct ttatatatct tgtggaaagg acgaaacacc ggtgtcagag
     3241 accggagttg tgttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa
     3301 cttgaaaaag tggcaccgag tcggtgcttt ttttcatggt catagctgtt tcctcatggt
     3361 aaaacgacgg ccagtgaggg cctatttccc atgattcctt catatttgca tatacgatac
     3421 aaggctgtta gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa
     3481 aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt
     3541 aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat
     3601 atcttgtgga aaggacgaaa caccgtataa gtaaggaacg cgcgcgtttt agagctagaa
     3661 atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg
     3721 ctttttttca tggtcatagc tgtttcctgg acgtaaaacg acggccagtg agggcctatt
     3781 tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga taattggaat
     3841 taatttgact gtaaacacaa agatattagt acaaaatacg tgacgtagaa agtaataatt
     3901 tcttgggtag tttgcagttt taaaattatg ttttaaaatg gactatcata tgcttaccgt
     3961 aacttgaaag tatttcgatt tcttggcttt atatatcttg tggaaaggac gaaacaccgg
     4021 ctgccatagc aacgatggag ttttagagct agaaatagca agttaaaata aggctagtcc
     4081 gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt ttcatggtca tagctgtttc
     4141 ctccaggtaa aacgacggcc agtgagggcc tatttcccat gattccttca tatttgcata
     4201 tacgatacaa ggctgttaga gagataattg gaattaattt gactgtaaac acaaagatat
     4261 tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat
     4321 tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg
     4381 ctttatatat cttgtggaaa ggacgaaaca ccgggttccc aaagcagagg gcggttttag
     4441 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg
     4501 agtcggtgct ttttttcatg gtcatagctg tttccttgtt gtaaaacgac ggccagtgag
     4561 ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt tagagagata
     4621 attggaatta atttgactgt aaacacaaag atattagtac aaaatacgtg acgtagaaag
     4681 taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga ctatcatatg
     4741 cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg gaaaggacga
     4801 aacaccggcg cgcgtagtta attcatggtt ttagagctag aaatagcaag ttaaaataag
     4861 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttt catggtcata
     4921 gctgtttcct aacgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct
     4981 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc caattccgat
     5041 catattcaat aacccttaat ataacttcgt ataatgtatg ctatacgaag ttattaggtc
     5101 tgaagaggag tttacgtcca gccaagctta ggatctcgac ctcgaaattc taccgggtag
     5161 gggaggcgct tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt
     5221 ggcgctacac aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac
     5281 cggctccgtt ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt
     5341 tcccccccgc cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca
     5401 ctagtctcgt gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca
     5461 gcggccaata gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt
     5521 ccgggggcgg gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag
     5581 gcccggcatt ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat
     5641 ctccgggcct ttcgacctgc atccatctag atctcgagca gctgaagctt accatgaccg
     5701 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
     5761 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
     5821 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     5881 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     5941 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     6001 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     6061 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     6121 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     6181 ccgcgcccca caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     6241 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc
     6301 cccacgaccc gcagcgcccg accgaaagga gcgcacgacc ccatgcatcg atgatctaga
     6361 gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     6421 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     6481 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     6541 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     6601 atggcttctg aggcggaaag aacctgcagc ccaagcttgg cgtaatcatg gtcatagctg
     6661 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     6721 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     6781 ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga tcccatagtg actggatatg
     6841 ttgtgtttta cagtattatg tagtctgttt tttatgcaaa atctaattta atatattgat
     6901 atttatatca ttttacgttt ctcgttcagc ttttttgtac aaacttgtgc tagcattcta
     6961 gttgatatct ataacaagaa aatatatata taataagtta tcacgtaagt agaacatgaa
     7021 ataacaatat aattatcgta tgagttaaat cttaaaagtc acgtaaaaga taatcatgcg
     7081 tcattttgac tcacgcggtc gttatagttc aaaatcagtg acacttaccg cattgacaag
     7141 cacgcctcac gggagctcca agcggcgact gagatgtcct aaatgcacag cgacggattc
     7201 gcgctattta gaaagagaga gcaatatttc aagaatgcat gcgtcaattt tacgcagact
     7261 atctttctag ggttaatcta gaatggcgcg ccccaattcg ccctatagtg agtcgtatta
     7321 cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     7381 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     7441 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag
     7501 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     7561 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     7621 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     7681 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     7741 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     7801 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     7861 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     7921 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
     7981 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     8041 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     8101 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     8161 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     8221 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     8281 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     8341 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     8401 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     8461 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     8521 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     8581 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     8641 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     8701 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     8761 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     8821 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     8881 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     8941 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     9001 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     9061 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     9121 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     9181 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     9241 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     9301 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     9361 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     9421 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     9481 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     9541 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     9601 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     9661 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     9721 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     9781 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     9841 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     9901 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     9961 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
    10021 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
    10081 ggaaacagct atgaccatga ttacgccaag cgcgcaatta accctcacta aagggaacaa
    10141 aagctggatt aattaagcgg ccgcatacgc gtatactaga ttaaccctag aaagataatc
    10201 atattgtgac gtacgttaaa gataatcatg cgtaaaattg acgcatgtgt tttatcggtc
    10261 tgtatatcga ggtttattta ttaatttgaa tagatattaa gttttattat atttacactt
    10321 acatactaat aataaattca acaaacaatt tatttatgtt tatttattta ttaaaaaaaa
    10381 acaaaaactc aaaatttctt ctataaagta acaaaacttt taactagagg atctcgagct
    10441 caagcttcga attccccctg taaaacgacg gccagtgagg gcctatttcc catgattcct
    10501 tcatatttgc atatacgata caaggctgtt agagagataa ttggaattaa tttgactgta
    10561 aacacaaaga tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt
    10621 gcagttttaa aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat
    10681 ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccgccca gcactttggg
    10741 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
    10801 ggcaccgagt cggtgctttt tttcatggtc atagctgttt cctcatggta aaacgacggc
    10861 cagtgagggc ctatttccca tgattccttc atatttgcat atacgataca aggctgttag
    10921 agagataatt ggaattaatt tgactgtaaa cacaaagata ttagtacaaa atacgtgacg
    10981 tagaaagtaa taatttcttg ggtagtttgc agttttaaaa ttatgtttta aaatggacta
    11041 tcatatgctt accgtaactt gaaagtattt cgatttcttg gctttatata tcttgtggaa
    11101 aggacgaaac accgaatccc agcactttgt tttagagcta gaaatagcaa gttaaaataa
    11161 ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt tcatggtcat
    11221 agctgtttcc tggacgtaaa acgacggcca gtgagggcct atttcccatg attccttcat
    11281 atttgcatat acgatacaag gctgttagag agataattgg aattaatttg actgtaaaca
    11341 caaagatatt agtacaaaat acgtgacgta gaaagtaata atttcttggg tagtttgcag
    11401 ttttaaaatt atgttttaaa atggactatc atatgcttac cgtaacttga aagtatttcg
    11461 atttcttggc tttatatatc ttgtggaaag gacgaaacac cggcctccca aagtgcgttt
    11521 tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga aaaagtggca
    11581 ccgagtcggt gctttttttc atggtcatag ctgtttcctc caggtaaaac gacggccagt
    11641 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
    11701 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
    11761 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
    11821 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
    11881 cgaaacaccg gctacttggg aggcgtttta gagctagaaa tagcaagtta aaataaggct
    11941 agtccgttat caacttgaaa aagtggcacc gagtcggtgc tttttttcat ggtcatagct
    12001 gtttccttgt tgtaaaacga cggccagtga gggcctattt cccatgattc cttcatattt
    12061 gcatatacga tacaaggctg ttagagagat aattggaatt aatttgactg taaacacaaa
    12121 gatattagta caaaatacgt gacgtagaaa gtaataattt cttgggtagt ttgcagtttt
    12181 aaaattatgt tttaaaatgg actatcatat gcttaccgta acttgaaagt atttcgattt
    12241 cttggcttta tatatcttgt ggaaaggacg aaacaccggc ctcccaagta gcgttttaga
    12301 gctagaaata gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga
    12361 gtcggtgctt tttttcatgg tcatagctgt ttcctaacga gcggccgc
//
