LOCUS       sequence-485140-       14095 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Improved Dox-inducible neurogenin-2; Piggybac Tet-ON plasmid for 
            differentiating hiPSCs into glutamatergic neurons via NGN2 
            expression. oNGN2 is a Phospho-mutant which avoids inhibition by cdk
            kinase..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14095)
  TITLE     Characterization of iPSC lines for the iPSC Neurodegenerative 
            Disease Initiative (iNDI)
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 14095)
  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: 
            "Unpublished"
COMMENT     Sequence Label: iC13 PB-TO-oNgn2-BFP-puro
FEATURES             Location/Qualifiers
     source          1..14095
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     351..370
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    369..424
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     470..489
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(580..1326)
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     primer_bind     complement(1358..1377)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          1385..2401
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(1439..1461)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(2402..2678)
                     /label=chicken beta-actin promoter
     enhancer        2680..3059
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     polyA_signal    3060..3108
                     /note="synthetic polyadenylation signal"
     misc_feature    3122..3213
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     3162..3181
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     protein_bind    complement(4425..4458)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        4459..4670
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          4671..5609
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5566..5586
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             5632..6228
                     /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(5632..5651)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6128..6148
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             6238..6291
                     /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             6298..6324
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     CDS             6340..6366
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     regulatory      6388..6397
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             6394..7104
                     /codon_start=1
                     /product="enhanced monomeric blue fluorescent protein 
                     (Subach et al., 2011)"
                     /label=mTagBFP2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRND
                     MALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     polyA_signal    7124..7348
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(7349..7382)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     repeat_region   7898..7932
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(8423..8439)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8447..8463
                     /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(8471..8477,8478..8495,8496..8501))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8516..8537
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(8825..9413)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8905..8924)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(9584..10375,10376..10444))
                     /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     10207..10226
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(10445..10549)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(10575..11030)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(10712..10733)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10924..10943
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     11157..11179
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     11171..11188
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     11172..11188
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   11318..11380
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        12485..12862
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    12492..12510
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    12528..12546
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    12564..12582
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    12600..12618
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    12636..12654
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    12672..12690
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    12708..12726
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     12771..12795
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     polyA_signal    13736..13857
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13773..13792)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13827..13846
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(13906..13922)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    13930..13946
                     /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(13954..13960,13961..13978,13979..13984))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13999..14020
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
       61 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggacgg ccgcacatgt
      121 aaggaaaatt ttagggatgt taaagaaaaa aataacacaa aacaaaatat aaaaaaaatc
      181 taacctcaag tcaaggcttt tctatggaat aaggaatgga cagcaggggg ctgtttcata
      241 tactgatgac ctctttatag ccaacctttg ttcatggcag ccagcatatg ggcatatgtt
      301 gccaaactct aaaccaaata ctcattctga tgttttaaat gatttgccct cccatatgtc
      361 cttccgagtg agagacacaa aaaattccaa cacactattg caatgaaaat aaatttcctt
      421 tattagccag aagtcagatg ctcaaggggc ttcatgatgt ccccataatt tttggcagag
      481 ggaaaaagat ctcagtggta tttgtgagcc agggcattgg ccacaccagt caccaccttc
      541 tgataggcag cctgcacctg aggagtgaat tcacgcgttt tagcctggca gcatatcgag
      601 atcgaagtca tcgagagcgt cagcagggag catatccaga tcaaaatcat ccagggcatc
      661 ggcgggcagc atgtccaggt cgaagtcgtc cagggcgtcg gtgggtccgc cgctctcgca
      721 cttcagctgc ttctcgaggc cgcagatgat cagttccagg ccgaacagga aggcgggctc
      781 ggcgccctgt ctgtcgaaca gctcgatggc ctgcttcagc agggggggca tgctgtcggt
      841 ggtgggtgtc tctctttcct ctttggcgac ctggtgttcc tgttcttcca gcacgcagcc
      901 cagggtgaag tggcccacgg cgctcagggc gtacagggcg ttttccaggc tgaagccctg
      961 ctggcacagg aaggccagct ggttttccag tgtctcgtac tgcttctcgg tgggtctggt
     1021 gcccaggtgc actttggcgc cgtcccggtg gctcagcagg gcgcatctgt agctcttggc
     1081 gttgttccgc aggaagtcct gccagctctc gccttccaga gggcagctgt gggtgtggtg
     1141 ccggtccagc atctcgatgg gcagggcgtc cagcagggcc cgcttgttct tcacgtgcca
     1201 gtacagggtg ggctgttcca cgcccagttt ctgggccagc ttccgggtgg tcaggccctc
     1261 gatgcccacg ccgttcagca gttccagggc gctgttgatc actttgctct tgtccagccg
     1321 gctcatgtcg acattctttg ccaaaatgat gagacagcac aataaccagc acgttgccca
     1381 ggagctgtag gaaaaagaag aaggcatgaa catggttagc agaggctcta gagccgccgg
     1441 tcacacgcca gaagccgaac cccgccctgc cccgtccccc ccgaaggcag ccgtcccccc
     1501 gcggacagcc ccgaggctgg agagggagaa ggggacggcg gcgcggcgac gcacgaaggc
     1561 cctccccgcc catttccttc ctgccggcgc cgcaccgctt cgccccgcgc ccgctagagg
     1621 gggtgcggcg gcgcctccca gatttcggct ccgcacagat ttgggacaaa ggaagtccct
     1681 gcgccctctc gcacgattac cataaaaggc aatggctgcg gctcgccgcg cctcgacagc
     1741 cgccggcgct ccgggggccg ccgcgcccct cccccgagcc ctccccggcc cgaggcggcc
     1801 ccgccccgcc cggcaccccc acctgccgcc accccccgcc cggcacggcg agccccgcgc
     1861 cacgccccgt acggagcccc gcacccgaag ccgggccgtg ctcagcaact cggggagggg
     1921 ggtgcagggg gggttgcagc ccgaccgacg cgcccacacc ccctgctcac ccccccacgc
     1981 acacaccccg cacgcagcct ttgttcccct cgcagccccc cccgcaccgc ggggcaccgc
     2041 ccccggccgc gctcccctcg cgcacactgc ggagcgcaca aagccccgcg ccgcgcccgc
     2101 agcgctcaca gccgccgggc agcgcggagc cgcacgcggc gctccccacg cacacacaca
     2161 cgcacgcacc ccccgagccg ctccccccgc acaaagggcc ctcccggagc ccctcaaggc
     2221 tttcacgcag ccacagaaaa gaaacaagcc gtcattaaac caagcgctaa ttacagcccg
     2281 gaggagaagg gccgtcccgc ccgctcacct gtgggagtaa cgcggtcagt cagagccggg
     2341 gcgggcggcg cgaggcggcg gcggagcggg gcacggggcg aaggcagcgc gcagcgactc
     2401 ccgcccgccg cgcgcttcgc tttttatagg gccgccgccg ccgccgcctc gccataaaag
     2461 gaaactttcg gagcgcgccg ctctgattgg ctgccgccgc acctctccgc ctcgccccgc
     2521 cccgcccctc gccccgcccc gccccgcctg gcgcgcgccc cccccccccc ccgcccccat
     2581 cgctgcacaa aataattaaa aaataaataa atacaaaatt gggggtgggg agggggggga
     2641 gatggggaga gtgaagcaga acgtggggct cacctcgacc atggtaatag cgatgactaa
     2701 tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca
     2761 ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg
     2821 atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt
     2881 ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt
     2941 gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg
     3001 gctatgaact aatgaccccg taattgatta ctattaataa ctagtcaata atcaatgtca
     3061 ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagtac
     3121 taacatacgc tctccatcaa aacaaaacga aacaaaacaa actagcaaaa taggctgtcc
     3181 ccagtgcaag tgcaggtgcc agaacatttc tctgagctca cggggacagc ccccccccaa
     3241 agcccccagg gatgtaatta cgtccctccc ccgctagggg gcagcagcga gccgcccggg
     3301 gctccgctcc ggtccggcgc tccccccgca tccccgagcc ggcagcgtgc ggggacagcc
     3361 cgggcacggg gaaggtggca cgggatcgct ttcctctgaa cgcttctcgc tgctctttga
     3421 gcctgcagac acctgggggg atacggggaa aaagctttag gctgaaagag agatttagaa
     3481 tgacagaatc atagaacggc ctgggttgca aaggagcaca gtgctcatcc agatccaacc
     3541 ccctgctatg tgcagggtca tcaaccagca gcccaggctg cccagagcca catccagcct
     3601 ggccttgaat gcctgcaggg atggggcatc cacagcctcc ttgggcaacc tgttcagtgc
     3661 gtcaccaccc tctgggggaa aaactgcctc ctcatatcca acccaaacct cccctgtctc
     3721 agtgtaaagc cattccccct tgtcctatca agggggagtt tgctgtgaca ttgttggtct
     3781 ggggtgacac atgtttgcca attcagtgca tcacggagag gcagatcttg gggataagga
     3841 agtgcaggac agcatggacg tgggacatgc aggtgttgag ggctctggga cactctccaa
     3901 gtcacagcgt tcagaacagc cttaaggata agaagatagg atagaaggac aaagagcaag
     3961 ttaaaaccca gcatggagag gagcacaaaa aggccacaga cactgctggt ccctgtgtct
     4021 gagcctgcat gtttgatggt gtctggatgc aagcagaagg ggtggaagag cttgcctgga
     4081 gagatacagc tgggtcagta ggactgggac aggcagctgg agaattgcca tgtagatgtt
     4141 catacaatcg tcaaatcatg aaggctggaa aagccctcca agatccccaa gaccaacccc
     4201 aacccaccca ccgtgcccac tggccatgtc cctcagtgcc acatccccac agttcttcat
     4261 cacctccagg gacggtgacc cccccacctc cgtgggcagc tgtgccactg cagcaccgct
     4321 ctttggagaa ggtaaatctt gctaaatcca gcccgaccct cccctggcac aacgtaaggc
     4381 cattatctct catccaactc caggacggag tcagtgagaa tattataact tcgtatagca
     4441 tacattatac gaagttatgg gcagagcgca catcgcccac agtccccgag aagttggggg
     4501 gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
     4561 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
     4621 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg
     4681 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta
     4741 cttccacctg gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg
     4801 agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc
     4861 ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt
     4921 tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc
     4981 aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg
     5041 cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag
     5101 cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg
     5161 gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag
     5221 ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga
     5281 cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt
     5341 cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt
     5401 agttctcgcg cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg
     5461 agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat
     5521 tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag
     5581 tggttcaaag tttttttctt ccatttcagg tgtcgtgatt taaatcccac catgaccgag
     5641 tacaagccca cggtgcgcct cgccacccgc gacgacgtcc ccagggccgt acgcaccctc
     5701 gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgatccgga ccgccacatc
     5761 gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag
     5821 gtgtgggtcg cggacgacgg cgccgccgtg gcggtctgga ccacgccgga gagcgtcgaa
     5881 gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg
     5941 gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg
     6001 ttcctggcca ccgtcggagt ctcgcccgac caccagggca agggtctggg cagcgccgtc
     6061 gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc
     6121 gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag
     6181 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgccgg atcgggagag
     6241 ggcagaggaa gtctgctaac atgcggtgac gtcgaggaga atcctggccc accggtccct
     6301 gccgctaaac gggtcaaact cgacggcggt tctggaggtc ctgctgcaaa gcgcgttaag
     6361 ttggatggag ggtcaggggg agccaccgcc accatggtgt ctaagggcga agagctgatt
     6421 aaggagaaca tgcacatgaa gctgtacatg gagggcaccg tggacaacca tcacttcaag
     6481 tgcacatccg agggcgaagg caagccctac gagggcaccc agaccatgag aatcaaggtg
     6541 gtcgagggcg gccctctccc cttcgccttc gacatcctgg ctactagctt cctctacggc
     6601 agcaagacct tcatcaacca cacccagggc atccccgact tcttcaagca gtccttccct
     6661 gagggcttca catgggagag agtcaccaca tacgaagacg ggggcgtgct gaccgctacc
     6721 caggacacca gcctccagga cggctgcctc atctacaacg tcaagatcag aggggtgaac
     6781 ttcacatcca acggccctgt gatgcagaag aaaacactcg gctgggaggc cttcaccgag
     6841 acgctgtacc ccgctgacgg cggcctggaa ggcagaaacg acatggccct gaagctcgtg
     6901 ggcgggagcc atctgatcgc aaacgccaag accacatata gatccaagaa acccgctaag
     6961 aacctcaaga tgcctggcgt ctactatgtg gactacagac tggaaagaat caaggaggcc
     7021 aacaacgaga cctacgtcga gcagcacgag gtggcagtgg ccagatactg cgacctccct
     7081 agcaaactgg ggcacaagct taattaggcg gccgctaaga attctgtgcc ttctagttgc
     7141 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     7201 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     7261 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     7321 catgctgggg atgcggtggg ctctatggat aacttcgtat agcatacatt atacgaagtt
     7381 atttggtatg gctttttccc cgtatccccc caggtgtctg caggctcaaa gagcagcgag
     7441 aagcgttcag aggaaagcga tcccgtgcca ccttccccgt gcccgggctg tccccgcacg
     7501 ctgccggctc ggggatgcgg ggggagcgcc ggaccggagc ggagccccgg gcggctcgct
     7561 gctgccccct agcgggggag ggacgtaatt acatccctgg gggctttggg ggggggctgt
     7621 ccctgattct ataacaagaa aatatatata taataagtta tcacgtaagt agaacatgaa
     7681 ataacaatat aattatcgta tgagttaaat cttaaaagtc acgtaaaaga taatcatgcg
     7741 tcattttgac tcacgcggtc gttatagttc aaaatcagtg acacttaccg cattgacaag
     7801 cacgcctcac gggagctcca agcggcgact gagatgtcct aaatgcacag cgacggattc
     7861 gcgctattta gaaagagaga gcaatatttc aagaatgcat gcgtcaattt tacgcagact
     7921 atctttctag ggttaatcta gctgcatcag gatcatatcg tcgggtcttt tttccggctc
     7981 agtcatcgcc caagctggcg ctatctgggc atcggggagg aagaagcccg tgccttttcc
     8041 cgcgaggttg aagcggcatg gaaagagttt gccgaggatg actgctgctg cattgacgtt
     8101 gagcgaaaac gcacgtttac catgatgatt cgggaaggtg tggccatgca cgcctttaac
     8161 ggtgaactgt tcgttcaggc cacctgggat accagttcgt cgcggctttt ccggacacag
     8221 ttccggatgg tcagcccgaa gcgcatcagc aacccgaaca ataccggcga cagccggaac
     8281 tgccgtgccg gtgtgcagat taatgacagc ggtgcggcgc tgggatatta cgtcagcgag
     8341 gacgggtatc ctggctggat gccgcagaaa tggacatgga taccccgtga gttacccggc
     8401 gggcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     8461 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     8521 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     8581 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     8641 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     8701 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     8761 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     8821 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     8881 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     8941 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     9001 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     9061 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     9121 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     9181 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     9241 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     9301 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     9361 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     9421 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     9481 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     9541 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     9601 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     9661 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     9721 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     9781 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     9841 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     9901 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     9961 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
    10021 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
    10081 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
    10141 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
    10201 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
    10261 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
    10321 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
    10381 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
    10441 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
    10501 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
    10561 gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
    10621 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
    10681 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
    10741 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
    10801 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
    10861 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
    10921 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
    10981 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc
    11041 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
    11101 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
    11161 gtcacgacgt tgtaaaacga cggccagtga gcgcgcctcg ttcattcacg tttttgaacc
    11221 cgtggaggac gggcagactc gcggtgcaaa tgtgttttac agcgtgatgg agcagatgaa
    11281 gatgctcgac acgctgcaga acacgcagct agattaaccc tagaaagata atcatattgt
    11341 gacgtacgtt aaagataatc atgtgtaaaa ttgacgcatg tgttttatcg gtctgtatat
    11401 cgaggtttat ttattaattt gaatagatat taagttttat tatatttaca cttacatact
    11461 aataataaat tcaacaaaca atttatttat gtttatttat ttattaaaaa aaacaaaaac
    11521 tcaaaatttc ttctataaag taacaaaact tttatgaggg acagcccccc cccaaagccc
    11581 ccagggatgt aattacgtcc ctcccccgct agggggcagc agcgagccgc ccggggctcc
    11641 gctccggtcc ggcgctcccc ccgcatcccc gagccggcag cgtgcgggga cagcccgggc
    11701 acggggaagg tggcacggga tcgctttcct ctgaacgctt ctcgctgctc tttgagcctg
    11761 cagacacctg gggggatacg gggaaaaggc ctccacggcc ttaattaagg gaggtggtcc
    11821 ctgcagttac gccaatgata acccccgcca gaaaaatctt agtagccttc cctttttgtt
    11881 ttccgtgccc caactcggcg gattgactcg gccccttccg gaaacacccg aatcaacttc
    11941 tagtcaaatt attgttcacg ccgcaatgac ccacccctgg cccgcgtctg tggaactgac
    12001 ccctggtgta caggagagtt cgctgctgaa agtggtccca aaggggtact agtttttaag
    12061 ctcccaactc cccctccccc agcgtctgga ggattccaca ccctcgcacc gcaggggcga
    12121 ggaagtgggc ggagtccggt tttggcgcca gccgctgagg ctgccaagca gaaaagccac
    12181 cgctgaggag actccggtca ctgtcctcgc cccgcctccc ccttccctcc ccttggggac
    12241 caccgggcgc cacgccgcga acggtaagtg ccgcggtcgt cggcgcctcc gccctccccc
    12301 tagggcccca attcccagcg ggcgcggcgc gcggcccctc cccccgccgg gcgcgcgccc
    12361 gctgccccgc ccttcgtggc cgcccggcgt gggcggtgcc acccctcccc ccggcggccc
    12421 cgcgcgcagc tcccggctcc ctcccccttc ggatgtggct tgagctgtag gcgcggaggg
    12481 taccagttta ctccctatca gtgatagaga acgtatgaag agtttactcc ctatcagtga
    12541 tagagaacgt atgcagactt tactccctat cagtgataga gaacgtataa ggagtttact
    12601 ccctatcagt gatagagaac gtatgaccag tttactccct atcagtgata gagaacgtat
    12661 ctacagttta ctccctatca gtgatagaga acgtatatcc agtttactcc ctatcagtga
    12721 tagagaacgt ataagcttta ggcgtgtacg gtgggcgcct ataaaagcag agctcgttta
    12781 gtgaaccgtc agatcgcctg gagcaattcc acaacacttt tgtcttatac caactttccg
    12841 taccacttcc taccctcgta aaggatccgc cgccaccatg tttgtgaagt ctgaaacact
    12901 ggaacttaaa gaagaggaag atgtcttggt tttgctgggc tctgcagcac cagcgctcgc
    12961 cgctttgacg cctttgagta gttccgctga tgaggaagaa gaagaagaac ccggagcatc
    13021 cggtggtgct cgccgccaaa gaggagcaga agctggacaa ggagctcgcg gcggtgtcgc
    13081 agccggggca gaaggttgta ggccagccag gttgctcggg ttggtgcatg actgtaagag
    13141 gagaccgagc agggcaaggg ctgtgagtag aggtgctaaa actgcagaaa ctgtccaaag
    13201 aattaagaag actagacgac ttaaagctaa taatagggaa cgcaatagga tgcataatct
    13261 gaatgccgcc ctcgatgccc ttagagaagt actgccaact tttcctgaag atgcaaaact
    13321 gacaaagatt gaaactctcc gttttgcaca taattatatt tgggctttga ctgaaacact
    13381 ccggcttgca gaccattgtg gtggtggcgg aggaggtctt ccaggcgccc tgtttagcga
    13441 agccgtcctc ttggcccctg gtggagcgtc agctgctctc tcttcctcag gcgatgcacc
    13501 ggccccagct tcaacctggt catgtacaaa tgccccagcc ccaagcagta gcgttagctc
    13561 taactcaaca gccccttata gttgtaccct cgcaccggca gcccctgcgg gtagtgatat
    13621 ggattactgg caacctcctc caccggataa acatcgttac gccccacatt tgccaatcgc
    13681 gcgagactgc attctgtaag ttaacgttta aactcgcgaa tcgatcgccc accccaactt
    13741 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
    13801 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
    13861 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
    13921 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
    13981 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
    14041 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgc
//
