LOCUS       sequence-416489-       17301 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NFIB and SOX9 mediated astrocye induction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 17301)
  AUTHORS   Lam I, Ndayisaba A, Lewis AJ, Fu Y, Sagredo GT, Zaccagnini L, Sandoe
            J, Sanz RL, Vahdatshoar A, Martin TD, Morshed N, Ichihashi T, 
            Tripathi A, Ramalingam N, Oettgen-Suazo C, Bartels T, Schabinger M, 
            Hallacli E, Jiang X, Verma A, Tea C, Wang Z, Hakozaki H, Yu X, Hyles
            K, Park C, Theunissen TW, Wang H, Jaenisch R, Lindquist S, Stevens 
            B, Stefanova N, Wenning G, Luk KC, Sanchez Pernaute R, Gomez-Esteban
            JC, Felsky D, Kiyota Y, Sahni N, Yi SS, Chung C, Stahlberg H, Ferrer
            I, Schoneberg J, Elledge SJ, Dettmer U, Halliday GM, Bartels T, 
            Khurana V
  TITLE     Rapid iPSC inclusionopathy models shed light on formation, 
            consequence and molecular subtype of alpha-synuclein inclusions
  JOURNAL   bioRxiv 2022.11.08.515615
REFERENCE   2  (bases 1 to 17301)
  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: "bioRxiv 
            2022.11.08.515615"
COMMENT     Sequence Label: piggyBac-rtTA (4th_Gen)-NFIB-SOX9-IRES-mKate2
            ...
FEATURES             Location/Qualifiers
     source          1..17301
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    19..495
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(218..237)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2254..2316
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    2440..2488
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2513..2532
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(2530..3660)
                     /codon_start=1
                     /product="herpes simplex virus thymidine kinase"
                     /label=HSV TK
                     /note="confers sensitivity to nucleoside analogs such as 
                     acyclovir or ganciclovir"
                     /translation="MASYPGHQHASAFDQAARSRGHSNRRTALRPRRQQEATEVRPEQK
                     MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWRVLGASETIANIYTTQ
                     HRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGSSHAPPPALTLIFDR
                     HPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGALPEDRHIDRLAKRQR
                     PGERLDLAMLAAIRRVYGLLANTVRYLQGGGSWREDWGQLSGTAVPPQGAEPQSNAGPR
                     PHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRPMHVFILDYDQSPAGCRDAL
                     LQLTSGMIQTHVTTPGSIPTICDLARTFAREMGEAN"
     promoter        complement(3667..4177)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4871..4891)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4871..4889)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4910..4926)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4910..4926)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4923..4945)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4934..4950
                     /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(4958..4964,4965..4982,4983..4988))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5003..5024
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5141..5158)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5312..5900)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5392..5411)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6071..6862,6863..6931))
                     /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     6694..6713
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6932..7036)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7111..7130)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     repeat_region   7311..7345
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    7898..8122
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     8111..8128
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(8151..8720)
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
     CDS             complement(8739..9095)
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALL"
     CDS             complement(9525..9545)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(9570..9719)
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     CDS             complement(10377..10430)
                     /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             complement(10440..10835)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     promoter        complement(10845..11355)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     protein_bind    12067..12085
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    12103..12121
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    12177..12195
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    12223..12241
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    12270..12288
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    12306..12324
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    12445..12469
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             13741..13797
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             15331..15372
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    complement(15380..15404)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    15456..16023
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    15473..16025
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(15617..15634)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     15844..15863
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             16043..16741
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     primer_bind     complement(16766..16786)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(17184..17195)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 gctgctcgag agatctacgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc
       61 ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt
      121 ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag
      181 gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt
      241 gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg
      301 cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg
      361 tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct
      421 caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct
      481 cccttccctg tccttgtacc ttaattaagt taacatgcat agtactgagc tcacggggac
      541 agcccccccc ccaaagcccc cagggatgta attacgtccc tcccccgcta gggggcagca
      601 gcgagccgcc cggggctccg ctccggtccg gcgctccccc cgcatccccg agccggcagc
      661 gtgcggggac agcccgggca cggggaaggt ggcacgggat cgctttcctc tgaacgcttc
      721 tcgctgctct ttgagcctgc agacacctgg ggggatacgg ggaaaaagct ttaggctgaa
      781 agagagattt agaatgacag aatcatagaa cggcctgggt tgcaaaggag cacagtgctc
      841 atccagatcc aaccccctgc tatgtgcagg gtcatcaacc agcagcccag gctgcccaga
      901 gccacatcca gcctggcctt gaatgcctgc agggatgggg catccacagc ctccttgggc
      961 aacctgttca gtgcgtcacc accctctggg ggaaaaactg cctcctcata tccaacccaa
     1021 acctcccctg tctcagtgta aagccattcc cccttgtcct atcaaggggg agtttgctgt
     1081 gacattgttg gtctggggtg acacatgttt gccaattcag tgcatcacgg agaggcagat
     1141 cttggggata aggaagtgca ggacagcatg gacgtgggac atgcaggtgt tgagggctct
     1201 gggacactct ccaagtcaca gcgttcagaa cagccttaag gataagaaga taggatagaa
     1261 ggacaaagag caagttaaaa cccagcatgg agaggagcac aaaaaggcca cagacactgc
     1321 tggtccctgt gtctgagcct gcatgtttga tggtgtctgg atgcaagcag aaggggtgga
     1381 agagcttgcc tggagagata cagctgggtc agtaggactg ggacaggcag ctggagaatt
     1441 gccatgtaga tgttcataca atcgtcaaat catgaaggct ggaaaagccc tccaagatcc
     1501 ccaagaccaa ccccaaccca cccaccgtgc ccactggcca tgtccctcag tgccacatcc
     1561 ccacagttct tcatcacctc cagggacggt gaccccccca cctccgtggg cagctgtgcc
     1621 actgcagcac cgctctttgg agaaggtaaa tcttgctaaa tccagcccga ccctcccctg
     1681 gcacaacgta aggccattat ctctcatcca actccaggac ggagtcagtg agaatattgt
     1741 ttaaacatta taggcgcgcc attgctggga ttacaggcgt gaaccactgc tcccttccct
     1801 gtccttttaa aagttttgtt actttatakm msmamagttc ttcatcacct ccagggacgg
     1861 tgaccccccc acctccgtgg gcagctgtgc cactgcagca ccgctctttg gagaaggtaa
     1921 atcttgctaa atccagcccg accctcccct ggcacaacgt aaggccatta tctctcatcc
     1981 aactccagga cggagtcagt gagaatattg tttaaacatt ataggcgcgc cattgctggg
     2041 attacaggcg tgaaccactg ctcccttccc tgtcctttta aaagttttgt tactttatag
     2101 aagaaatttt gagtttttgt ttttttttaa taaataaata aacataaata aattgtttgt
     2161 tgaatttatt attagtatgt aagtgtaaat ataataaaac ttaatatcta ttcaaattaa
     2221 taaataaacc tcgatataca gaccgataaa acacatgcgt caattttacg catgattatc
     2281 tttaacgtac gtcacaatat gattatcttt ctagggttaa tctagtatac gcgtatgcgg
     2341 ccgctgggga aaaggaagaa acgcgggcgt attggcccca atggggtctc ggtggggtat
     2401 cgacagagtg ccagccctgg gaccgaaccc cgcgtttatg aacaaacgac ccaacacccg
     2461 tgcgttttat tctgtctttt tattgccgtc atagcgcggg ttccttccgg tattgtctcc
     2521 ttccgtgttt cagttagcct cccccatctc ccgggcaaac gtgcgcgcca ggtcgcagat
     2581 cgtcggtatg gagcctgggg tggtgacgtg ggtctggatc atcccggagg taagttgcag
     2641 cagggcgtcc cggcagccgg cgggcgattg gtcgtaatcc aggataaaga cgtgcatggg
     2701 acggaggcgt ttggccaaga cgtccaaggc ccaggcaaac acgttgtaca ggtcgccgtt
     2761 gggggccagc aactcggggg cccgaaacag ggtaaataac gtgtccccga tatggggtcg
     2821 tgggcccgcg ttgctctggg gctcggcacc ctggggcggc acggccgtcc ccgaaagctg
     2881 tccccaatcc tcccgccacg acccgccgcc ctgcagatac cgcaccgtat tggcaagcag
     2941 cccgtaaacg cggcgaatcg cggccagcat agccaggtca agccgctcgc cggggcgctg
     3001 gcgtttggcc aggcggtcga tgtgtctgtc ctccggaagg gcccccaaca cgatgtttgt
     3061 gccgggcaag gtcggcggga tgagggccac gaacgccagc acggcctggg gggtcatgct
     3121 gcccataagg tatcgcgcgg ccgggtagca caggagggcg gcgatgggat ggcggtcgaa
     3181 gatgagggtg agggccgggg gcggggcatg tgagctccca gcctcccccc cgatatgagg
     3241 agccagaacg gcgtcggtca cggcataagg catgcccatt gttatctggg cgcttgtcat
     3301 taccaccgcc gcgtccccgg ccgatatctc accctggtca aggcggtgtt gtgtggtgta
     3361 gatgttcgcg attgtctcgg aagcccccag cacccgccag taagtcatcg gctcgggtac
     3421 gtagacgata tcgtcgcgcg aacccagggc caccagcagt tgcgtggtgg tggttttccc
     3481 catcccgtgg ggaccgtcta tataaacccg cagtagcgtg ggcattttct gctccgggcg
     3541 gacttccgtg gcttcttgct gccggcgagg gcgcaacgcc gtacgtcggt tgctatggcc
     3601 gcgagaacgc gcagcctggt cgaacgcaga cgcgtgctga tggccggggt acgaagccat
     3661 ggcggtctgg ggagagaggt cggtgattcg gtcaacgagg gagccgactg ccgacgtgcg
     3721 ctccggaggc ttgcagaatg cggaacaccg cgcgggcagg aacagggccc acactaccgc
     3781 cccacacccc gcctcccgca ccgccccttc ccggccgctg ctctcggcgc gccctgctga
     3841 gcagccgcta ttggccacag cccatcgcgg tcggcgcgct gccattgctc cctggcgctg
     3901 tccgtctgcg agggtactag tgagacgtgc ggcttccgtt tgtcacgtcc ggcacgccgc
     3961 gaaccgcaag gaaccttccc gacttagggg cggagcagga agcgtcgccg gggggcccac
     4021 aagggtagcg gcgaagatcc gggtgacgct gcgaacggac gtgaagaatg tgcgagaccc
     4081 agggtcggcg ccgctgcgtt tcccggaacc acgcccagag cagccgcgtc cctgcgcaaa
     4141 cccagggctg ccttggaaaa ggcgcaaccc caacccccct ccgcgcctac agctcaagcc
     4201 acatccgaag ggggagggag ccgggagctg cgcgcggggc cgccgggggg aggggtggca
     4261 ccgcccacgc cgggcggcca cgaagggcgg ggcagcgggc gcgcgcccgg cggggggagg
     4321 ggccgcgcgc cgcgcccgct gggaattggg gccctagggg gagggcggag gcgccgacga
     4381 ccgcggcact taccgttcgc ggcgtggcgc ccggtggtcc ccaaggggag ggaaggggga
     4441 ggcggggcga ggacagtgac cggagtctcc tcagcggtgg cttttctgct tggcagcctc
     4501 agcggctggc gccaaaaccg gactccgccc acttcctcgc ccctgcggtg cgagggtgtg
     4561 gaatcctcca gacgctgggg gagggggagt tgggagctta aaaactagta cccctttggg
     4621 accactttca gcagcgaact ctcctgtaca ccaggggtca gttccacaga cgcgggccag
     4681 gggtgggtca ttgcggcgtg aacaataatt tgactagaag ttgattcggg tgtttccgga
     4741 aggggccgag tcaatccgcc gagttggggc acggaaaaca aaaagggaag gctactaaga
     4801 tttttctggc gggggttatc attggcgtaa ctgcagggac cacctccctt aattaatcca
     4861 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt
     4921 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4981 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     5041 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     5101 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     5161 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5221 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5281 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5341 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5401 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5461 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5521 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5581 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5641 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5701 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5761 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5821 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5881 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5941 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6001 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6061 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6121 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6181 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6241 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6301 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6361 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6421 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6481 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6541 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6601 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6661 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6721 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6781 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6841 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6901 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6961 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7021 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     7081 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     7141 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
     7201 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
     7261 ttttgcgctg cttcggggag atgggggagg ctaactgagc tctagattaa ccctagaaag
     7321 atagtctgcg taaaattgac gcatgcattc ttgaaatatt gctctctctt tctaaatagc
     7381 gcgaatccgt cgctgtgcat ttaggacatc tcagtcgccg cttggagctc ccgtgaggcg
     7441 tgcttgtcaa tgcggtaagt gtcactgatt ttgaactata acgaccgcgt gagtcaaaat
     7501 gacgcatgat tatcttttac gtgactttta agatttaact catacgataa ttatattgtt
     7561 atttcatgtt ctacttacgt gataacttat tatatatata ttttcttgtt atagatatca
     7621 caaatgcttg tccttccttc ctgtaacaca gccattaaac caggagcatc gcccttcccc
     7681 ggccctcagg taagaggacc aaataccgta gccgtttcca atttcagtcc tttagcgcca
     7741 cctggtgcta actactctat cacgctttta tccaataact acctttgtaa atttcctttc
     7801 aaaagttctg gccgggcgcg gtggctcacg cttgtaatcc cagcactttg tgaggggtca
     7861 ggagttcgag acgatcaact agaatgaatg tcgagaccca tagagcccac cgcatcccca
     7921 gcatgcctgc tattgtcttc ccaatcctcc cccttgctgt cctgccccac cccacccccc
     7981 agaatagaat gacacctact cagacaatgc gatgcaattt cctcatttta ttaggaaagg
     8041 acagtgggag tggcaccttc cagggtcaag gaaggcacgg gggaggggca aacaacagat
     8101 ggctggcaac tagaaggcac agtcgaggct gatcagcggg tttaaactca aaacagagat
     8161 gtgtcgaaga tggacagtcc tgtgctgata tgcatggcat gcaagaggca ctcgtcgttc
     8221 aggaaggtat ccagaatctc gttcaattcc ggggtcaggg gtgagtccag gttcagatcc
     8281 tcggtcatgg actcaagtgt ggttgtcagc tcatccagat ggccccttgg gggcggatgg
     8341 gaaaggtcca tttggccaca gattgcagcc tcttccttct ggggaatcac agtatcggcc
     8401 atctcccgaa gggctttgac agcctggctc gtctcttcat cgggatcctc caacaggtga
     8461 ctggcctcgg gagtcactgc gggcgctgga tccagtggct gagggactgg tgccggggtc
     8521 agtgacccga ctggctcatg tactggaccg gttggtgttg gtgcgaggct ggcggggagt
     8581 gggcggttgg cccatggact tcctggagga tgaaatggcc ggattcgttt tggctggcac
     8641 acctcgcggc cctcaaacac gtcactaata gcggagccgg cctcaggctt cggcaaaaac
     8701 atcccttccc tggaatcccg gctgccgctg ccagagccca gcaaggctga gaaatccata
     8761 tcggcgatag agctgaagtc ctcgtcgcca gacagcagtc cattaggcag gcctggtgct
     8821 cccagagggg caggagctgg atcaggaggc ctctgagcgc ctgtcacgag ccgggtgatg
     8881 gcctcggggt attccatcag catgggctcg gtggtgtgag gggccacagg gatgccctgg
     8941 ttcagcagct gctggaactc gctgttgtcc acgctggcca ggtcggtgaa cacggcagga
     9001 tcggtgctgt ttcccagcag ggctcccaga tcctcgtcgt cgaactgcag ctgcagcaga
     9061 gcttcagaca gtgtgccctc gccggcctgt gtaggtttag gggctggtgg agccacagcc
     9121 tgtggaggtc caggagccag cacaggcacg ggtgctggtg cctgagccag tgcagacacc
     9181 atggctggag ctggtgcagg agcaggagcc tgaggcagca cctgaggagg ggctggagcc
     9241 agagcagagg cctgagagat ctggccgctg gggaacacca tggtagggaa ctcgtcgtag
     9301 ttgatggtgc tcaggctgct ggtgaagggg taaggctggg gggcaggttt tggcacgctg
     9361 gcgctggatc tgctgggcac ggcgattctt ctaggtggag gtctagggtc ggtggggccg
     9421 ctgaaggggg acttcttcat gatgctcttg aatgtctcgt aggtccgctt ccgcttttcc
     9481 tcgatccggt gccggtcgtc ggtgtcgggc aggtactggc taccaacttt gcgtttcttt
     9541 ttcggagatc cggaacttct agagttaatc agcatgtcca ggtcgaaatc atcaagggcg
     9601 tcactgccga gcatgtcgag gtcaaagtca tcaagggcat ccgaaccaag catgtcaagg
     9661 tcaaaatcat cgagggcgtc acttcccagc atatccagat caaaatcgtc caatgcgtca
     9721 gcccgtccgg aaccgctggc ctccggcccg ccgctttcgc actttagctg tttctccagg
     9781 ccacatatga ttagttccag gccgaaaagg aaggcaggtt cggctccctg ccggtcgaac
     9841 agctcaattg cttgtttcag aagtgggggc atagaatcgg tggtaggtgt ctctctttcc
     9901 tcttttgcta cttgatgctc ctgttcctcc aatacgcagc ccagtgtaaa gtggcccacg
     9961 gcggacagag cgtacagtgc gttctccagg gagaagcctt gctgacacag gaacgcgagc
    10021 tgattttcca gggtttcgta ctgtttctct gttgggcggg tgccgagatg cactttagcc
    10081 ccgtcgcgat gtgagaggag agcacagcgg tatgacttgg cgttgttccg cagaaagtct
    10141 tgccatgact cgccttccag ggggcaggag tgggtatgat gcctgtccag catctcgatt
    10201 ggcagggcat cgagcagggc ccgcttgttc ttcacgtgcc agtacagggt aggctgctca
    10261 actcccagct tttgagcgag tttccttgtc gtcaggcctt cgataccgac tccattgagt
    10321 aattccagag cagagtttat gactttgctc ttgtccagtc tagacataga accacctgga
    10381 ccagggtttt cttcaacatc accacaagtg aggagagaac ctctaccttc gccggatccg
    10441 ccctcccaca cataaccaga gggcagcaat tcacgaatcc caactgccgt cggctgtcca
    10501 tcactgtcct tcactatggc tttgatccca ggatgcagat cgagaagcac ctgtcggcac
    10561 cgtccgcagg ggctcaagat gcccctgttc tcatttccga tcgcgacgat acaagtcagg
    10621 ttgccagctg ccgcagcagc agcagtgccc agcaccacga gttctgcaca aggtccccca
    10681 gtaaaatgat atacattgac accagtgaag atgcggccgt cgctagagag agctgcgctg
    10741 gcgacgctgt agtcttcaga gatggggatg ctgttgattg tagccgttgc tctttcaatg
    10801 agggtggatt cttcttgaga caaaggcttg gccatggtga attgctgggg agagaggtcg
    10861 gtgattcggt caacgaggga gccgactgcc gacgtgcgct ccggaggctt gcagaatgcg
    10921 gaacaccgcg cgggcaggaa cagggcccac actaccgccc cacaccccgc ctcccgcacc
    10981 gccccttccc ggccgctgct ctcggcgcgc cctgctgagc agccgctatt ggccacagcc
    11041 catcgcggtc ggcgcgctgc cattgctccc tggcgctgtc cgtctgcgag ggtactagtg
    11101 agacgtgcgg cttccgtttg tcacgtccgg cacgccgcga accgcaagga accttcccga
    11161 cttaggggcg gagcaggaag cgtcgccggg gggcccacaa gggtagcggc gaagatccgg
    11221 gtgacgctgc gaacggacgt gaagaatgtg cgagacccag ggtcggcgcc gctgcgtttc
    11281 ccggaaccac gcccagagca gccgcgtccc tgcgcaaacc cagggctgcc ttggaaaagg
    11341 cgcaacccca accccgggga ggtggtccct gcagttacgc caatgataac ccccgccaga
    11401 aaaatcttag tagccttccc tttttgtttt ccgtgcccca actcggcgga ttgactcggc
    11461 cccttccgga aacacccgaa tcaacttcta gtcaaattat tgttcacgcc gcaatgaccc
    11521 acccctggcc cgcgtctgtg gaactgaccc ctggtgtaca ggagagttcg ctgctgaaag
    11581 tggtcccaaa ggggtactag tttttaagct cccaactccc cctcccccag cgtctggagg
    11641 attccacacc ctcgcaccgc aggggcgagg aagtgggcgg agtccggttt tggcgccagc
    11701 cgctgaggct gccaagcaga aaagccaccg ctgaggagac tccggtcact gtcctcgccc
    11761 cgcctccccc ttccctcccc ttggggacca ccgggcgcca cgccgcgaac ggtaagtgcc
    11821 gcggtcgtcg gcgcctccgc cctcccccta gggccccaat tcccagcggg cgcggcgcgc
    11881 ggcccctccc cccgccgggc gcgcgcccgc tgccccgccc ttcgtggccg cccggcgtgg
    11941 gcggtgccac ccctcccccc ggcggccccg cgcgcagctc ccggctccct cccccttcgg
    12001 atgtggcttg agctgtaggc gcggaggcta gctggggatc tggtgggctc tattggtaga
    12061 ctttagtccc tatcagtgat agagaaggta tctccagttt actccctatc agtgatagag
    12121 acatccatgg aatcaacttc agctttatga gtaagaagcc ctataggatg tggctttccc
    12181 tatcagtgat agagaactaa tgataagacc actgcacttt agtccctatc agtgatagag
    12241 atgagatctt ctggtgtaca ggagttttct ccctatcagt gatagagagt ctgtgtgcca
    12301 tttgatccct atcagtgata gagactgtat ttccaggtag gcctgtacag tgggaggcct
    12361 atataagcag agctccttta gtgaaccctc agatggccag agataagaga gaaaagaaga
    12421 gtaagaagaa atataagagc caccacaagt ttgtacaaaa aagcaggctg catgatgtat
    12481 tctcccatct gtctcactca ggatgaattt cacccattca tcgaggcact tcttccacat
    12541 gtccgtgcaa ttgcctatac ttggttcaac ctgcaggctc gaaaacgcaa gtactttaaa
    12601 aagcatgaga agcgaatgtc aaaggatgaa gaaagagcag tcaaagatga gcttctcagt
    12661 gaaaagcctg aaatcaaaca gaagtgggca tccaggctcc ttgccaaact gcgcaaagat
    12721 attcgccagg agtatcgaga ggactttgtg ctcaccgtga ctggcaagaa gcacccgtgc
    12781 tgtgtcttat ccaatcccga ccagaagggt aagattagga gaatcgactg cctgcgacag
    12841 gcagacaaag tctggcgtct ggatctagtc atggtgatcc tgttcaaagg catccccttg
    12901 gaaagtaccg atggagagcg gctcatgaaa tccccacatt gcacaaaccc agcactttgt
    12961 gtccagccac atcatatcac agtatcagtt aaggagcttg atttgttttt ggcatactac
    13021 gtgcaggagc aagattctgg acaatcagga agtccaagcc acaatgatcc tgccaagaat
    13081 cctccaggtt accttgagga tagttttgta aaatctggag tcttcaatgt atcagaactt
    13141 gtaagagtat ccagaacgcc cataacccag ggaactggag tcaacttccc aattggagaa
    13201 atcccaagcc aaccatacta tcatgacatg aactcggggg tcaatcttca gaggtctctg
    13261 tcttctccac caagcagcaa aagacccaaa actatatcca tagatgaaaa tatggaacca
    13321 agtcctacag gagactttta cccctctcca agttcaccag ctgctggaag tcgaacatgg
    13381 cacgaaagag atcaagatat gtcttctccg actactatga agaagcctga aaagccattg
    13441 ttcagctctg catctccaca ggattcttcc ccaagactga gcactttccc ccagcaccac
    13501 catcccggaa tacctggagt tgcacacagt gtcatctcaa ctcgaactcc acctccacct
    13561 tcaccgttgc catttccaac acaagctatc cttcctccag ccccatcgag ctacttttct
    13621 catccaacaa tcagatatcc tccccacctg aatcctcagg atactctgaa gaactatgta
    13681 ccttcttatg acccatccag tccacaaacc agccagtcct ggtacctggg cggtagcggt
    13741 gctactaact tctctctcct gaaacaagca ggcgacgtag aggagaatcc aggtcccatg
    13801 aatctcctgg accccttcat gaagatgacc gacgagcagg agaagggcct gtccggcgcc
    13861 cccagcccca ccatgtccga ggactccgcg ggctcgccct gcccgtcggg ctccggctcg
    13921 gacaccgaga acacgcggcc ccaggagaac acgttcccca agggcgagcc cgatctgaag
    13981 aaggagagcg aggaggacaa gttccccgtg tgcatccgcg aggcggtcag ccaggtgctc
    14041 aaaggctacg actggacgct ggtgcccatg ccggtgcgcg tcaacggctc cagcaagaac
    14101 aagccgcacg tcaagcggcc catgaacgcc ttcatggtgt gggcgcaggc ggcgcgcagg
    14161 aagctcgcgg accagtaccc gcacttgcac aacgccgagc tcagcaagac gctgggcaag
    14221 ctctggagac ttctgaacga gagcgagaag cggcccttcg tggaggaggc ggagcggctg
    14281 cgcgtgcagc acaagaagga ccacccggat tacaagtacc agccgcggcg gaggaagtcg
    14341 gtgaagaacg ggcaggcgga ggcagaggag gccacggagc agacgcacat ctcccccaac
    14401 gccatcttca aggcgctgca ggccgactcg ccacactcct cctccggcat gagcgaggtg
    14461 cactcccccg gcgagcactc ggggcaatcc cagggcccac cgaccccacc caccaccccc
    14521 aaaaccgacg tgcagccggg caaggctgac ctgaagcgag aggggcgccc cttgccagag
    14581 gggggcagac agccccctat cgacttccgc gacgtggaca tcggcgagct gagcagcgac
    14641 gtcatctcca acatcgagac cttcgatgtc aacgagtttg accagtacct gccgcccaac
    14701 ggccacccgg gggtgccggc cacgcacggc caggtcacct acacgggcag ctacggcatc
    14761 agcagcaccg cggccacccc ggcgagcgcg ggccacgtgt ggatgtccaa gcagcaggcg
    14821 ccgccgccac ccccgcagca gcccccacag gccccgccgg ccccgcaggc gcccccgcag
    14881 ccgcaggcgg cgcccccaca gcagccggcg gcacccccgc agcagccaca ggcgcacacg
    14941 ctgaccacgc tgagcagcga gccgggccag tcccagcgaa cgcacatcaa gacggagcag
    15001 ctgagcccca gccactacag cgagcagcag cagcactcgc cccaacagat cgcctacagc
    15061 cccttcaacc tcccacacta cagcccctcc tacccgccca tcacccgctc acagtacgac
    15121 tacaccgacc accagaactc cagctcctac tacagccacg cggcaggcca gggcaccggc
    15181 ctctactcca ccttcaccta catgaacccc gctcagcgcc ccatgtacac ccccatcgcc
    15241 gacacctctg gggtcccttc catcccgcag acccacagcc cccagcactg ggaacaaccc
    15301 gtctacacac agctcactcg acctggatcc ggtaagccta tccctaaccc tctcctcggt
    15361 ctcgattcta cgtagtagta cccagctttc ttgtacaaag tggttgatct gctggaagat
    15421 ggcgattaga gaggtacgaa gatggcgatt agagatccct cccccccccc taacgttact
    15481 ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata
    15541 ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt
    15601 cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa
    15661 gcagttcctc tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag
    15721 cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca
    15781 cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc
    15841 aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat
    15901 tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta
    15961 aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga
    16021 taatatggcc acacaaatgg tgatggtgag cgagctgatt aaggagaaca tgcacatgaa
    16081 gctgtacatg gagggcaccg tgaacaacca ccacttcaag tgcacatccg agggcgaagg
    16141 caagccctac gagggcaccc agaccatgag aatcaaggcg gtcgagggcg gccctctccc
    16201 cttcgccttc gacatcctgg ctaccagctt catgtacggc agcaaaacct tcatcaacca
    16261 cacccagggc atccccgact tctttaagca gtccttcccc gagggcttca catgggagag
    16321 agtcaccaca tacgaagacg ggggcgtgct gaccgctacc caggacacca gcctccagga
    16381 cggctgcctc atctacaacg tcaagatcag aggggtgaac ttcccatcca acggccctgt
    16441 gatgcagaag aaaacactcg gctgggaggc ctccaccgag accctgtacc ccgctgacgg
    16501 cggcctggaa ggcagagccg acatggccct gaagctcgtg ggcgggggcc acctgatctg
    16561 caacttgaag accacataca gatccaagaa acccgctaag aacctcaaga tgcccggcgt
    16621 ctactatgtg gacagaagac tggaaagaat caaggaggcc gacaaagaga cctacgtcga
    16681 gcagcacgag gtggctgtgg ccagatactg cgacctccct agcaaactgg ggcacagata
    16741 aacgcgatca tcgtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
    16801 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
    16861 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
    16921 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
    16981 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
    17041 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
    17101 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg
    17161 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
    17221 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
    17281 ccctttgggc cgcctccccg c
//
