LOCUS       sequence-496201-       15684 bp    DNA     circular SYN 06-JAN-2026
DEFINITION  Ngn2 mediated cortical neuron induction, along with sfGFP 
            over-expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15684)
  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 15684)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 6, 2026 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)-sfGFP-IRES-NGN2-puro
            (...
FEATURES             Location/Qualifiers
     source          1..15684
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1..25)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    77..644
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    94..646
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(238..255)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     465..484
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             658..681
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1474..1527
                     /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             1534..2133
                     /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(1534..1553)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2030..2050
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    2134..2722
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2187..2207)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2605..2616)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2737..3213
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2936..2955)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4701..4763
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    4887..4935
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4960..4979
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4977..6107)
                     /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(6114..6624)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7318..7338)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7318..7336)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7357..7373)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7357..7373)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7370..7392)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7381..7397
                     /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(7405..7411,7412..7429,7430..7435))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7450..7471
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7588..7605)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7759..8347)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7839..7858)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8518..9309,9310..9378))
                     /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     9141..9160
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9379..9483)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9558..9577)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     repeat_region   9758..9792
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    10345..10569
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     10558..10575
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(10598..11167)
                     /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(11186..11542)
                     /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(11972..11992)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(12017..12166)
                     /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(12824..12877)
                     /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(12887..13282)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     promoter        complement(13292..13802)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     protein_bind    14514..14532
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    14550..14568
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    14624..14642
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    14670..14688
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    14717..14735
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    14753..14771
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    14892..14916
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             14970..15683
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
ORIGIN
        1 acccagcttt cttgtacaaa gtggttgatc tgctggaaga tggcgattag agaggtacga
       61 agatggcgat tagagatccc tccccccccc ctaacgttac tggccgaagc cgcttggaat
      121 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
      181 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
      241 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
      301 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
      361 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
      421 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
      481 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
      541 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
      601 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaatggac
      661 tacaaggacg acgatgacaa gttcgtcaaa tctgagactc tggagttgaa ggaggaagag
      721 gaggtactga tgctgctggg ctcggcttcc ccggcctcgg cgaccctgac cccgatgtcc
      781 tccagcgcgg acgaggagga ggacgaggag ctgcgccggc cgggctccgc gcgtgggcag
      841 cgtggagcgg aagccgggca gggggtgcag ggcagtccgg cgtcgggtgc cgggggttgc
      901 cggccagggc ggctgctggg cctgatgcac gagtgcaagc gtcgcccgtc gcgctcacgg
      961 gccgtctccc gaggtgccaa gacggcggag acggtgcagc gcatcaagaa gacccgcagg
     1021 ctcaaggcca acaaccgcga gcgcaaccgc atgcacaacc taaacgccgc gctggacgcg
     1081 ctgcgcgagg tgctgcccac cttccccgag gatgccaagc tcacgaagat cgagacgctg
     1141 cgcttcgccc acaattacat ctgggcgctc accgagactc tgcgcctggc ggaccactgc
     1201 gccggcgccg gtggcctcca gggggcgctc ttcacggagg cggtgctcct gagcccggga
     1261 gctgcgctcg gcgccagcgg ggacagccct tctccacctt cctcctggag ctgcaccaac
     1321 agcccggcgt catcctccaa ctccacgtcc ccatacagct gcactttatc gcccgctagc
     1381 cccgggtcag acgtggacta ctggcagccc ccacctccgg agaagcatcg ttatgcgcct
     1441 cacctgcccc tcgccaggga ctgtatctct agagagggca ggggaagtct tctaacatgc
     1501 ggggacgtgg aggaaaatcc cggcccactc gagatgaccg agtacaagcc cacggtgcgc
     1561 ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac
     1621 taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt caccgagctg
     1681 caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac
     1741 ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc
     1801 gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca gcaacagatg
     1861 gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc
     1921 gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag
     1981 gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc
     2041 ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc
     2101 acctggtgca tgacccgcaa gcccggtgcc tgaaatcaac ctctggatta caaaatttgt
     2161 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     2221 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     2281 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     2341 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     2401 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     2461 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     2521 tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc
     2581 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     2641 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     2701 tccctttggg ccgcctcccc gcctcgagag atctacgggt ggcatccctg tgacccctcc
     2761 ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata
     2821 aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg
     2881 ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat
     2941 tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg
     3001 gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc
     3061 aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg
     3121 gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta
     3181 caggcgtgaa ccactgctcc cttccctgtc cttgtacctt aattaagtta acatgcatag
     3241 tactgagctc acggggacag cccccccccc aaagccccca gggatgtaat tacgtccctc
     3301 ccccgctagg gggcagcagc gagccgcccg gggctccgct ccggtccggc gctccccccg
     3361 catccccgag ccggcagcgt gcggggacag cccgggcacg gggaaggtgg cacgggatcg
     3421 ctttcctctg aacgcttctc gctgctcttt gagcctgcag acacctgggg ggatacgggg
     3481 aaaaagcttt aggctgaaag agagatttag aatgacagaa tcatagaacg gcctgggttg
     3541 caaaggagca cagtgctcat ccagatccaa ccccctgcta tgtgcagggt catcaaccag
     3601 cagcccaggc tgcccagagc cacatccagc ctggccttga atgcctgcag ggatggggca
     3661 tccacagcct ccttgggcaa cctgttcagt gcgtcaccac cctctggggg aaaaactgcc
     3721 tcctcatatc caacccaaac ctcccctgtc tcagtgtaaa gccattcccc cttgtcctat
     3781 caagggggag tttgctgtga cattgttggt ctggggtgac acatgtttgc caattcagtg
     3841 catcacggag aggcagatct tggggataag gaagtgcagg acagcatgga cgtgggacat
     3901 gcaggtgttg agggctctgg gacactctcc aagtcacagc gttcagaaca gccttaagga
     3961 taagaagata ggatagaagg acaaagagca agttaaaacc cagcatggag aggagcacaa
     4021 aaaggccaca gacactgctg gtccctgtgt ctgagcctgc atgtttgatg gtgtctggat
     4081 gcaagcagaa ggggtggaag agcttgcctg gagagataca gctgggtcag taggactggg
     4141 acaggcagct ggagaattgc catgtagatg ttcatacaat cgtcaaatca tgaaggctgg
     4201 aaaagccctc caagatcccc aagaccaacc ccaacccacc caccgtgccc actggccatg
     4261 tccctcagtg ccacatcccc acagttcttc atcacctcca gggacggtga cccccccacc
     4321 tccgtgggca gctgtgccac tgcagcaccg ctctttggag aaggtaaatc ttgctaaatc
     4381 cagcccgacc ctcccctggc acaacgtaag gccattatct ctcatccaac tccaggacgg
     4441 agtcagtgag aatattgttt aaacattata ggcgcgccat tgctgggatt acaggcgtga
     4501 accactgctc ccttccctgt ccttttaaaa gttttgttac tttatagaag aaattttgag
     4561 tttttgtttt tttttaataa ataaataaac ataaataaat tgtttgttga atttattatt
     4621 agtatgtaag tgtaaatata ataaaactta atatctattc aaattaataa ataaacctcg
     4681 atatacagac cgataaaaca catgcgtcaa ttttacgcat gattatcttt aacgtacgtc
     4741 acaatatgat tatctttcta gggttaatct agtatacgcg tatgcggccg ctggggaaaa
     4801 ggaagaaacg cgggcgtatt ggccccaatg gggtctcggt ggggtatcga cagagtgcca
     4861 gccctgggac cgaaccccgc gtttatgaac aaacgaccca acacccgtgc gttttattct
     4921 gtctttttat tgccgtcata gcgcgggttc cttccggtat tgtctccttc cgtgtttcag
     4981 ttagcctccc ccatctcccg ggcaaacgtg cgcgccaggt cgcagatcgt cggtatggag
     5041 cctggggtgg tgacgtgggt ctggatcatc ccggaggtaa gttgcagcag ggcgtcccgg
     5101 cagccggcgg gcgattggtc gtaatccagg ataaagacgt gcatgggacg gaggcgtttg
     5161 gccaagacgt ccaaggccca ggcaaacacg ttgtacaggt cgccgttggg ggccagcaac
     5221 tcgggggccc gaaacagggt aaataacgtg tccccgatat ggggtcgtgg gcccgcgttg
     5281 ctctggggct cggcaccctg gggcggcacg gccgtccccg aaagctgtcc ccaatcctcc
     5341 cgccacgacc cgccgccctg cagataccgc accgtattgg caagcagccc gtaaacgcgg
     5401 cgaatcgcgg ccagcatagc caggtcaagc cgctcgccgg ggcgctggcg tttggccagg
     5461 cggtcgatgt gtctgtcctc cggaagggcc cccaacacga tgtttgtgcc gggcaaggtc
     5521 ggcgggatga gggccacgaa cgccagcacg gcctgggggg tcatgctgcc cataaggtat
     5581 cgcgcggccg ggtagcacag gagggcggcg atgggatggc ggtcgaagat gagggtgagg
     5641 gccgggggcg gggcatgtga gctcccagcc tcccccccga tatgaggagc cagaacggcg
     5701 tcggtcacgg cataaggcat gcccattgtt atctgggcgc ttgtcattac caccgccgcg
     5761 tccccggccg atatctcacc ctggtcaagg cggtgttgtg tggtgtagat gttcgcgatt
     5821 gtctcggaag cccccagcac ccgccagtaa gtcatcggct cgggtacgta gacgatatcg
     5881 tcgcgcgaac ccagggccac cagcagttgc gtggtggtgg ttttccccat cccgtgggga
     5941 ccgtctatat aaacccgcag tagcgtgggc attttctgct ccgggcggac ttccgtggct
     6001 tcttgctgcc ggcgagggcg caacgccgta cgtcggttgc tatggccgcg agaacgcgca
     6061 gcctggtcga acgcagacgc gtgctgatgg ccggggtacg aagccatggc ggtctgggga
     6121 gagaggtcgg tgattcggtc aacgagggag ccgactgccg acgtgcgctc cggaggcttg
     6181 cagaatgcgg aacaccgcgc gggcaggaac agggcccaca ctaccgcccc acaccccgcc
     6241 tcccgcaccg ccccttcccg gccgctgctc tcggcgcgcc ctgctgagca gccgctattg
     6301 gccacagccc atcgcggtcg gcgcgctgcc attgctccct ggcgctgtcc gtctgcgagg
     6361 gtactagtga gacgtgcggc ttccgtttgt cacgtccggc acgccgcgaa ccgcaaggaa
     6421 ccttcccgac ttaggggcgg agcaggaagc gtcgccgggg ggcccacaag ggtagcggcg
     6481 aagatccggg tgacgctgcg aacggacgtg aagaatgtgc gagacccagg gtcggcgccg
     6541 ctgcgtttcc cggaaccacg cccagagcag ccgcgtccct gcgcaaaccc agggctgcct
     6601 tggaaaaggc gcaaccccaa cccccctccg cgcctacagc tcaagccaca tccgaagggg
     6661 gagggagccg ggagctgcgc gcggggccgc cggggggagg ggtggcaccg cccacgccgg
     6721 gcggccacga agggcggggc agcgggcgcg cgcccggcgg ggggaggggc cgcgcgccgc
     6781 gcccgctggg aattggggcc ctagggggag ggcggaggcg ccgacgaccg cggcacttac
     6841 cgttcgcggc gtggcgcccg gtggtcccca aggggaggga agggggaggc ggggcgagga
     6901 cagtgaccgg agtctcctca gcggtggctt ttctgcttgg cagcctcagc ggctggcgcc
     6961 aaaaccggac tccgcccact tcctcgcccc tgcggtgcga gggtgtggaa tcctccagac
     7021 gctgggggag ggggagttgg gagcttaaaa actagtaccc ctttgggacc actttcagca
     7081 gcgaactctc ctgtacacca ggggtcagtt ccacagacgc gggccagggg tgggtcattg
     7141 cggcgtgaac aataatttga ctagaagttg attcgggtgt ttccggaagg ggccgagtca
     7201 atccgccgag ttggggcacg gaaaacaaaa agggaaggct actaagattt ttctggcggg
     7261 ggttatcatt ggcgtaactg cagggaccac ctcccttaat taatccagct tttgttccct
     7321 ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     7381 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     7441 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     7501 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     7561 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     7621 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     7681 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     7741 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     7801 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     7861 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     7921 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     7981 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     8041 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     8101 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     8161 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     8221 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     8281 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     8341 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     8401 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     8461 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     8521 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     8581 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     8641 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     8701 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     8761 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     8821 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     8881 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     8941 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     9001 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     9061 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     9121 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     9181 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     9241 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     9301 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     9361 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     9421 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     9481 gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     9541 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     9601 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     9661 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
     9721 cggggagatg ggggaggcta actgagctct agattaaccc tagaaagata gtctgcgtaa
     9781 aattgacgca tgcattcttg aaatattgct ctctctttct aaatagcgcg aatccgtcgc
     9841 tgtgcattta ggacatctca gtcgccgctt ggagctcccg tgaggcgtgc ttgtcaatgc
     9901 ggtaagtgtc actgattttg aactataacg accgcgtgag tcaaaatgac gcatgattat
     9961 cttttacgtg acttttaaga tttaactcat acgataatta tattgttatt tcatgttcta
    10021 cttacgtgat aacttattat atatatattt tcttgttata gatatcacaa atgcttgtcc
    10081 ttccttcctg taacacagcc attaaaccag gagcatcgcc cttccccggc cctcaggtaa
    10141 gaggaccaaa taccgtagcc gtttccaatt tcagtccttt agcgccacct ggtgctaact
    10201 actctatcac gcttttatcc aataactacc tttgtaaatt tcctttcaaa agttctggcc
    10261 gggcgcggtg gctcacgctt gtaatcccag cactttgtga ggggtcagga gttcgagacg
    10321 atcaactaga atgaatgtcg agacccatag agcccaccgc atccccagca tgcctgctat
    10381 tgtcttccca atcctccccc ttgctgtcct gccccacccc accccccaga atagaatgac
    10441 acctactcag acaatgcgat gcaatttcct cattttatta ggaaaggaca gtgggagtgg
    10501 caccttccag ggtcaaggaa ggcacggggg aggggcaaac aacagatggc tggcaactag
    10561 aaggcacagt cgaggctgat cagcgggttt aaactcaaaa cagagatgtg tcgaagatgg
    10621 acagtcctgt gctgatatgc atggcatgca agaggcactc gtcgttcagg aaggtatcca
    10681 gaatctcgtt caattccggg gtcaggggtg agtccaggtt cagatcctcg gtcatggact
    10741 caagtgtggt tgtcagctca tccagatggc cccttggggg cggatgggaa aggtccattt
    10801 ggccacagat tgcagcctct tccttctggg gaatcacagt atcggccatc tcccgaaggg
    10861 ctttgacagc ctggctcgtc tcttcatcgg gatcctccaa caggtgactg gcctcgggag
    10921 tcactgcggg cgctggatcc agtggctgag ggactggtgc cggggtcagt gacccgactg
    10981 gctcatgtac tggaccggtt ggtgttggtg cgaggctggc ggggagtggg cggttggccc
    11041 atggacttcc tggaggatga aatggccgga ttcgttttgg ctggcacacc tcgcggccct
    11101 caaacacgtc actaatagcg gagccggcct caggcttcgg caaaaacatc ccttccctgg
    11161 aatcccggct gccgctgcca gagcccagca aggctgagaa atccatatcg gcgatagagc
    11221 tgaagtcctc gtcgccagac agcagtccat taggcaggcc tggtgctccc agaggggcag
    11281 gagctggatc aggaggcctc tgagcgcctg tcacgagccg ggtgatggcc tcggggtatt
    11341 ccatcagcat gggctcggtg gtgtgagggg ccacagggat gccctggttc agcagctgct
    11401 ggaactcgct gttgtccacg ctggccaggt cggtgaacac ggcaggatcg gtgctgtttc
    11461 ccagcagggc tcccagatcc tcgtcgtcga actgcagctg cagcagagct tcagacagtg
    11521 tgccctcgcc ggcctgtgta ggtttagggg ctggtggagc cacagcctgt ggaggtccag
    11581 gagccagcac aggcacgggt gctggtgcct gagccagtgc agacaccatg gctggagctg
    11641 gtgcaggagc aggagcctga ggcagcacct gaggaggggc tggagccaga gcagaggcct
    11701 gagagatctg gccgctgggg aacaccatgg tagggaactc gtcgtagttg atggtgctca
    11761 ggctgctggt gaaggggtaa ggctgggggg caggttttgg cacgctggcg ctggatctgc
    11821 tgggcacggc gattcttcta ggtggaggtc tagggtcggt ggggccgctg aagggggact
    11881 tcttcatgat gctcttgaat gtctcgtagg tccgcttccg cttttcctcg atccggtgcc
    11941 ggtcgtcggt gtcgggcagg tactggctac caactttgcg tttctttttc ggagatccgg
    12001 aacttctaga gttaatcagc atgtccaggt cgaaatcatc aagggcgtca ctgccgagca
    12061 tgtcgaggtc aaagtcatca agggcatccg aaccaagcat gtcaaggtca aaatcatcga
    12121 gggcgtcact tcccagcata tccagatcaa aatcgtccaa tgcgtcagcc cgtccggaac
    12181 cgctggcctc cggcccgccg ctttcgcact ttagctgttt ctccaggcca catatgatta
    12241 gttccaggcc gaaaaggaag gcaggttcgg ctccctgccg gtcgaacagc tcaattgctt
    12301 gtttcagaag tgggggcata gaatcggtgg taggtgtctc tctttcctct tttgctactt
    12361 gatgctcctg ttcctccaat acgcagccca gtgtaaagtg gcccacggcg gacagagcgt
    12421 acagtgcgtt ctccagggag aagccttgct gacacaggaa cgcgagctga ttttccaggg
    12481 tttcgtactg tttctctgtt gggcgggtgc cgagatgcac tttagccccg tcgcgatgtg
    12541 agaggagagc acagcggtat gacttggcgt tgttccgcag aaagtcttgc catgactcgc
    12601 cttccagggg gcaggagtgg gtatgatgcc tgtccagcat ctcgattggc agggcatcga
    12661 gcagggcccg cttgttcttc acgtgccagt acagggtagg ctgctcaact cccagctttt
    12721 gagcgagttt ccttgtcgtc aggccttcga taccgactcc attgagtaat tccagagcag
    12781 agtttatgac tttgctcttg tccagtctag acatagaacc acctggacca gggttttctt
    12841 caacatcacc acaagtgagg agagaacctc taccttcgcc ggatccgccc tcccacacat
    12901 aaccagaggg cagcaattca cgaatcccaa ctgccgtcgg ctgtccatca ctgtccttca
    12961 ctatggcttt gatcccagga tgcagatcga gaagcacctg tcggcaccgt ccgcaggggc
    13021 tcaagatgcc cctgttctca tttccgatcg cgacgataca agtcaggttg ccagctgccg
    13081 cagcagcagc agtgcccagc accacgagtt ctgcacaagg tcccccagta aaatgatata
    13141 cattgacacc agtgaagatg cggccgtcgc tagagagagc tgcgctggcg acgctgtagt
    13201 cttcagagat ggggatgctg ttgattgtag ccgttgctct ttcaatgagg gtggattctt
    13261 cttgagacaa aggcttggcc atggtgaatt gctggggaga gaggtcggtg attcggtcaa
    13321 cgagggagcc gactgccgac gtgcgctccg gaggcttgca gaatgcggaa caccgcgcgg
    13381 gcaggaacag ggcccacact accgccccac accccgcctc ccgcaccgcc ccttcccggc
    13441 cgctgctctc ggcgcgccct gctgagcagc cgctattggc cacagcccat cgcggtcggc
    13501 gcgctgccat tgctccctgg cgctgtccgt ctgcgagggt actagtgaga cgtgcggctt
    13561 ccgtttgtca cgtccggcac gccgcgaacc gcaaggaacc ttcccgactt aggggcggag
    13621 caggaagcgt cgccgggggg cccacaaggg tagcggcgaa gatccgggtg acgctgcgaa
    13681 cggacgtgaa gaatgtgcga gacccagggt cggcgccgct gcgtttcccg gaaccacgcc
    13741 cagagcagcc gcgtccctgc gcaaacccag ggctgccttg gaaaaggcgc aaccccaacc
    13801 ccggggaggt ggtccctgca gttacgccaa tgataacccc cgccagaaaa atcttagtag
    13861 ccttcccttt ttgttttccg tgccccaact cggcggattg actcggcccc ttccggaaac
    13921 acccgaatca acttctagtc aaattattgt tcacgccgca atgacccacc cctggcccgc
    13981 gtctgtggaa ctgacccctg gtgtacagga gagttcgctg ctgaaagtgg tcccaaaggg
    14041 gtactagttt ttaagctccc aactccccct cccccagcgt ctggaggatt ccacaccctc
    14101 gcaccgcagg ggcgaggaag tgggcggagt ccggttttgg cgccagccgc tgaggctgcc
    14161 aagcagaaaa gccaccgctg aggagactcc ggtcactgtc ctcgccccgc ctcccccttc
    14221 cctccccttg gggaccaccg ggcgccacgc cgcgaacggt aagtgccgcg gtcgtcggcg
    14281 cctccgccct ccccctaggg ccccaattcc cagcgggcgc ggcgcgcggc ccctcccccc
    14341 gccgggcgcg cgcccgctgc cccgcccttc gtggccgccc ggcgtgggcg gtgccacccc
    14401 tccccccggc ggccccgcgc gcagctcccg gctccctccc ccttcggatg tggcttgagc
    14461 tgtaggcgcg gaggctagct ggggatctgg tgggctctat tggtagactt tagtccctat
    14521 cagtgataga gaaggtatct ccagtttact ccctatcagt gatagagaca tccatggaat
    14581 caacttcagc tttatgagta agaagcccta taggatgtgg ctttccctat cagtgataga
    14641 gaactaatga taagaccact gcactttagt ccctatcagt gatagagatg agatcttctg
    14701 gtgtacagga gttttctccc tatcagtgat agagagtctg tgtgccattt gatccctatc
    14761 agtgatagag actgtatttc caggtaggcc tgtacagtgg gaggcctata taagcagagc
    14821 tcctttagtg aaccctcaga tggccagaga taagagagaa aagaagagta agaagaaata
    14881 taagagccac cacaagtttg tacaaaaaag caggcttcgc cgccatgcag agcaccgtgc
    14941 ccagagccag ggacccacct gtggccacca gcaaaggcga ggaactcttt acgggggtag
    15001 taccgattct tgtggagttg gacggtgatg tcaatggtca caagttttct gtccgaggcg
    15061 aaggggaggg cgatgcgacc aatggtaagc ttacattgaa gttcatttgt acgacaggca
    15121 agctgccagt accttggcca acgttggtga cgacacttac atacggagta cagtgcttca
    15181 gccgctatcc tgatcacatg aaacgacatg atttttttaa atccgcaatg cccgaagggt
    15241 acgtgcagga acggacaatt tcctttaaag acgacgggac atataagaca agggcggagg
    15301 tgaagtttga aggcgacact cttgtgaata ggatagaact caaaggcatt gattttaaag
    15361 aagatgggaa cattttgggt cataagcttg agtataactt caattcccac aacgtctaca
    15421 taacagccga caaacaaaaa aacggaataa aagcaaactt caaaattagg cataacgttg
    15481 aggatggttc cgtccagctc gccgatcatt atcagcagaa tacgcctatt ggagatgggc
    15541 cggttctcct ccccgataat cactatctga gtacccagag tgtgctttct aaagatccca
    15601 acgaaaagcg cgatcatatg gttctcctcg aatttgtcac tgcagcaggg attacccatg
    15661 gtatggacga attgtataaa tgaa
//
