LOCUS       sequence-417157-       15956 bp    DNA     circular SYN 15-DEC-2025
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 15956)
  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 15956)
  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)-sfGFP-IRES-NGN2-puro
            (...
FEATURES             Location/Qualifiers
     source          1..15956
                     /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   4973..5035
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    5159..5207
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5232..5251
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(5249..6379)
                     /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(6386..6896)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7590..7610)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7590..7608)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7629..7645)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7629..7645)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7642..7664)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7653..7669
                     /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(7677..7683,7684..7701,7702..7707))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7722..7743
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7860..7877)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8031..8619)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8111..8130)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8790..9581,9582..9650))
                     /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     9413..9432
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9651..9755)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9830..9849)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     repeat_region   10030..10064
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    10617..10841
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     10830..10847
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(10870..11439)
                     /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(11458..11814)
                     /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(12244..12264)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(12289..12438)
                     /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(13096..13149)
                     /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(13159..13554)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     promoter        complement(13564..14074)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     protein_bind    14786..14804
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    14822..14840
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    14896..14914
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    14942..14960
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    14989..15007
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    15025..15043
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    15164..15188
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             15242..15955
                     /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 ccttttamma kktyystcag tgccacatcc ccacagttct
     4561 tcatcacctc cagggacggt gaccccccca cctccgtggg cagctgtgcc actgcagcac
     4621 cgctctttgg agaaggtaaa tcttgctaaa tccagcccga ccctcccctg gcacaacgta
     4681 aggccattat ctctcatcca actccaggac ggagtcagtg agaatattgt ttaaacatta
     4741 taggcgcgcc attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttttaa
     4801 aagttttgtt actttataga agaaattttg agtttttgtt tttttttaat aaataaataa
     4861 acataaataa attgtttgtt gaatttatta ttagtatgta agtgtaaata taataaaact
     4921 taatatctat tcaaattaat aaataaacct cgatatacag accgataaaa cacatgcgtc
     4981 aattttacgc atgattatct ttaacgtacg tcacaatatg attatctttc tagggttaat
     5041 ctagtatacg cgtatgcggc cgctggggaa aaggaagaaa cgcgggcgta ttggccccaa
     5101 tggggtctcg gtggggtatc gacagagtgc cagccctggg accgaacccc gcgtttatga
     5161 acaaacgacc caacacccgt gcgttttatt ctgtcttttt attgccgtca tagcgcgggt
     5221 tccttccggt attgtctcct tccgtgtttc agttagcctc ccccatctcc cgggcaaacg
     5281 tgcgcgccag gtcgcagatc gtcggtatgg agcctggggt ggtgacgtgg gtctggatca
     5341 tcccggaggt aagttgcagc agggcgtccc ggcagccggc gggcgattgg tcgtaatcca
     5401 ggataaagac gtgcatggga cggaggcgtt tggccaagac gtccaaggcc caggcaaaca
     5461 cgttgtacag gtcgccgttg ggggccagca actcgggggc ccgaaacagg gtaaataacg
     5521 tgtccccgat atggggtcgt gggcccgcgt tgctctgggg ctcggcaccc tggggcggca
     5581 cggccgtccc cgaaagctgt ccccaatcct cccgccacga cccgccgccc tgcagatacc
     5641 gcaccgtatt ggcaagcagc ccgtaaacgc ggcgaatcgc ggccagcata gccaggtcaa
     5701 gccgctcgcc ggggcgctgg cgtttggcca ggcggtcgat gtgtctgtcc tccggaaggg
     5761 cccccaacac gatgtttgtg ccgggcaagg tcggcgggat gagggccacg aacgccagca
     5821 cggcctgggg ggtcatgctg cccataaggt atcgcgcggc cgggtagcac aggagggcgg
     5881 cgatgggatg gcggtcgaag atgagggtga gggccggggg cggggcatgt gagctcccag
     5941 cctccccccc gatatgagga gccagaacgg cgtcggtcac ggcataaggc atgcccattg
     6001 ttatctgggc gcttgtcatt accaccgccg cgtccccggc cgatatctca ccctggtcaa
     6061 ggcggtgttg tgtggtgtag atgttcgcga ttgtctcgga agcccccagc acccgccagt
     6121 aagtcatcgg ctcgggtacg tagacgatat cgtcgcgcga acccagggcc accagcagtt
     6181 gcgtggtggt ggttttcccc atcccgtggg gaccgtctat ataaacccgc agtagcgtgg
     6241 gcattttctg ctccgggcgg acttccgtgg cttcttgctg ccggcgaggg cgcaacgccg
     6301 tacgtcggtt gctatggccg cgagaacgcg cagcctggtc gaacgcagac gcgtgctgat
     6361 ggccggggta cgaagccatg gcggtctggg gagagaggtc ggtgattcgg tcaacgaggg
     6421 agccgactgc cgacgtgcgc tccggaggct tgcagaatgc ggaacaccgc gcgggcagga
     6481 acagggccca cactaccgcc ccacaccccg cctcccgcac cgccccttcc cggccgctgc
     6541 tctcggcgcg ccctgctgag cagccgctat tggccacagc ccatcgcggt cggcgcgctg
     6601 ccattgctcc ctggcgctgt ccgtctgcga gggtactagt gagacgtgcg gcttccgttt
     6661 gtcacgtccg gcacgccgcg aaccgcaagg aaccttcccg acttaggggc ggagcaggaa
     6721 gcgtcgccgg ggggcccaca agggtagcgg cgaagatccg ggtgacgctg cgaacggacg
     6781 tgaagaatgt gcgagaccca gggtcggcgc cgctgcgttt cccggaacca cgcccagagc
     6841 agccgcgtcc ctgcgcaaac ccagggctgc cttggaaaag gcgcaacccc aacccccctc
     6901 cgcgcctaca gctcaagcca catccgaagg gggagggagc cgggagctgc gcgcggggcc
     6961 gccgggggga ggggtggcac cgcccacgcc gggcggccac gaagggcggg gcagcgggcg
     7021 cgcgcccggc ggggggaggg gccgcgcgcc gcgcccgctg ggaattgggg ccctaggggg
     7081 agggcggagg cgccgacgac cgcggcactt accgttcgcg gcgtggcgcc cggtggtccc
     7141 caaggggagg gaagggggag gcggggcgag gacagtgacc ggagtctcct cagcggtggc
     7201 ttttctgctt ggcagcctca gcggctggcg ccaaaaccgg actccgccca cttcctcgcc
     7261 cctgcggtgc gagggtgtgg aatcctccag acgctggggg agggggagtt gggagcttaa
     7321 aaactagtac ccctttggga ccactttcag cagcgaactc tcctgtacac caggggtcag
     7381 ttccacagac gcgggccagg ggtgggtcat tgcggcgtga acaataattt gactagaagt
     7441 tgattcgggt gtttccggaa ggggccgagt caatccgccg agttggggca cggaaaacaa
     7501 aaagggaagg ctactaagat ttttctggcg ggggttatca ttggcgtaac tgcagggacc
     7561 acctccctta attaatccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
     7621 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     7681 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     7741 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     7801 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     7861 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     7921 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     7981 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     8041 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     8101 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     8161 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     8221 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     8281 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     8341 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     8401 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     8461 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     8521 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     8581 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     8641 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     8701 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     8761 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     8821 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     8881 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     8941 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     9001 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     9061 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     9121 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     9181 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     9241 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     9301 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     9361 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     9421 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     9481 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     9541 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     9601 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     9661 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     9721 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     9781 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
     9841 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     9901 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     9961 atctgcttag ggttaggcgt tttgcgctgc ttcggggaga tgggggaggc taactgagct
    10021 ctagattaac cctagaaaga tagtctgcgt aaaattgacg catgcattct tgaaatattg
    10081 ctctctcttt ctaaatagcg cgaatccgtc gctgtgcatt taggacatct cagtcgccgc
    10141 ttggagctcc cgtgaggcgt gcttgtcaat gcggtaagtg tcactgattt tgaactataa
    10201 cgaccgcgtg agtcaaaatg acgcatgatt atcttttacg tgacttttaa gatttaactc
    10261 atacgataat tatattgtta tttcatgttc tacttacgtg ataacttatt atatatatat
    10321 tttcttgtta tagatatcac aaatgcttgt ccttccttcc tgtaacacag ccattaaacc
    10381 aggagcatcg cccttccccg gccctcaggt aagaggacca aataccgtag ccgtttccaa
    10441 tttcagtcct ttagcgccac ctggtgctaa ctactctatc acgcttttat ccaataacta
    10501 cctttgtaaa tttcctttca aaagttctgg ccgggcgcgg tggctcacgc ttgtaatccc
    10561 agcactttgt gaggggtcag gagttcgaga cgatcaacta gaatgaatgt cgagacccat
    10621 agagcccacc gcatccccag catgcctgct attgtcttcc caatcctccc ccttgctgtc
    10681 ctgccccacc ccacccccca gaatagaatg acacctactc agacaatgcg atgcaatttc
    10741 ctcattttat taggaaagga cagtgggagt ggcaccttcc agggtcaagg aaggcacggg
    10801 ggaggggcaa acaacagatg gctggcaact agaaggcaca gtcgaggctg atcagcgggt
    10861 ttaaactcaa aacagagatg tgtcgaagat ggacagtcct gtgctgatat gcatggcatg
    10921 caagaggcac tcgtcgttca ggaaggtatc cagaatctcg ttcaattccg gggtcagggg
    10981 tgagtccagg ttcagatcct cggtcatgga ctcaagtgtg gttgtcagct catccagatg
    11041 gccccttggg ggcggatggg aaaggtccat ttggccacag attgcagcct cttccttctg
    11101 gggaatcaca gtatcggcca tctcccgaag ggctttgaca gcctggctcg tctcttcatc
    11161 gggatcctcc aacaggtgac tggcctcggg agtcactgcg ggcgctggat ccagtggctg
    11221 agggactggt gccggggtca gtgacccgac tggctcatgt actggaccgg ttggtgttgg
    11281 tgcgaggctg gcggggagtg ggcggttggc ccatggactt cctggaggat gaaatggccg
    11341 gattcgtttt ggctggcaca cctcgcggcc ctcaaacacg tcactaatag cggagccggc
    11401 ctcaggcttc ggcaaaaaca tcccttccct ggaatcccgg ctgccgctgc cagagcccag
    11461 caaggctgag aaatccatat cggcgataga gctgaagtcc tcgtcgccag acagcagtcc
    11521 attaggcagg cctggtgctc ccagaggggc aggagctgga tcaggaggcc tctgagcgcc
    11581 tgtcacgagc cgggtgatgg cctcggggta ttccatcagc atgggctcgg tggtgtgagg
    11641 ggccacaggg atgccctggt tcagcagctg ctggaactcg ctgttgtcca cgctggccag
    11701 gtcggtgaac acggcaggat cggtgctgtt tcccagcagg gctcccagat cctcgtcgtc
    11761 gaactgcagc tgcagcagag cttcagacag tgtgccctcg ccggcctgtg taggtttagg
    11821 ggctggtgga gccacagcct gtggaggtcc aggagccagc acaggcacgg gtgctggtgc
    11881 ctgagccagt gcagacacca tggctggagc tggtgcagga gcaggagcct gaggcagcac
    11941 ctgaggaggg gctggagcca gagcagaggc ctgagagatc tggccgctgg ggaacaccat
    12001 ggtagggaac tcgtcgtagt tgatggtgct caggctgctg gtgaaggggt aaggctgggg
    12061 ggcaggtttt ggcacgctgg cgctggatct gctgggcacg gcgattcttc taggtggagg
    12121 tctagggtcg gtggggccgc tgaaggggga cttcttcatg atgctcttga atgtctcgta
    12181 ggtccgcttc cgcttttcct cgatccggtg ccggtcgtcg gtgtcgggca ggtactggct
    12241 accaactttg cgtttctttt tcggagatcc ggaacttcta gagttaatca gcatgtccag
    12301 gtcgaaatca tcaagggcgt cactgccgag catgtcgagg tcaaagtcat caagggcatc
    12361 cgaaccaagc atgtcaaggt caaaatcatc gagggcgtca cttcccagca tatccagatc
    12421 aaaatcgtcc aatgcgtcag cccgtccgga accgctggcc tccggcccgc cgctttcgca
    12481 ctttagctgt ttctccaggc cacatatgat tagttccagg ccgaaaagga aggcaggttc
    12541 ggctccctgc cggtcgaaca gctcaattgc ttgtttcaga agtgggggca tagaatcggt
    12601 ggtaggtgtc tctctttcct cttttgctac ttgatgctcc tgttcctcca atacgcagcc
    12661 cagtgtaaag tggcccacgg cggacagagc gtacagtgcg ttctccaggg agaagccttg
    12721 ctgacacagg aacgcgagct gattttccag ggtttcgtac tgtttctctg ttgggcgggt
    12781 gccgagatgc actttagccc cgtcgcgatg tgagaggaga gcacagcggt atgacttggc
    12841 gttgttccgc agaaagtctt gccatgactc gccttccagg gggcaggagt gggtatgatg
    12901 cctgtccagc atctcgattg gcagggcatc gagcagggcc cgcttgttct tcacgtgcca
    12961 gtacagggta ggctgctcaa ctcccagctt ttgagcgagt ttccttgtcg tcaggccttc
    13021 gataccgact ccattgagta attccagagc agagtttatg actttgctct tgtccagtct
    13081 agacatagaa ccacctggac cagggttttc ttcaacatca ccacaagtga ggagagaacc
    13141 tctaccttcg ccggatccgc cctcccacac ataaccagag ggcagcaatt cacgaatccc
    13201 aactgccgtc ggctgtccat cactgtcctt cactatggct ttgatcccag gatgcagatc
    13261 gagaagcacc tgtcggcacc gtccgcaggg gctcaagatg cccctgttct catttccgat
    13321 cgcgacgata caagtcaggt tgccagctgc cgcagcagca gcagtgccca gcaccacgag
    13381 ttctgcacaa ggtcccccag taaaatgata tacattgaca ccagtgaaga tgcggccgtc
    13441 gctagagaga gctgcgctgg cgacgctgta gtcttcagag atggggatgc tgttgattgt
    13501 agccgttgct ctttcaatga gggtggattc ttcttgagac aaaggcttgg ccatggtgaa
    13561 ttgctgggga gagaggtcgg tgattcggtc aacgagggag ccgactgccg acgtgcgctc
    13621 cggaggcttg cagaatgcgg aacaccgcgc gggcaggaac agggcccaca ctaccgcccc
    13681 acaccccgcc tcccgcaccg ccccttcccg gccgctgctc tcggcgcgcc ctgctgagca
    13741 gccgctattg gccacagccc atcgcggtcg gcgcgctgcc attgctccct ggcgctgtcc
    13801 gtctgcgagg gtactagtga gacgtgcggc ttccgtttgt cacgtccggc acgccgcgaa
    13861 ccgcaaggaa ccttcccgac ttaggggcgg agcaggaagc gtcgccgggg ggcccacaag
    13921 ggtagcggcg aagatccggg tgacgctgcg aacggacgtg aagaatgtgc gagacccagg
    13981 gtcggcgccg ctgcgtttcc cggaaccacg cccagagcag ccgcgtccct gcgcaaaccc
    14041 agggctgcct tggaaaaggc gcaaccccaa ccccggggag gtggtccctg cagttacgcc
    14101 aatgataacc cccgccagaa aaatcttagt agccttccct ttttgttttc cgtgccccaa
    14161 ctcggcggat tgactcggcc ccttccggaa acacccgaat caacttctag tcaaattatt
    14221 gttcacgccg caatgaccca cccctggccc gcgtctgtgg aactgacccc tggtgtacag
    14281 gagagttcgc tgctgaaagt ggtcccaaag gggtactagt ttttaagctc ccaactcccc
    14341 ctcccccagc gtctggagga ttccacaccc tcgcaccgca ggggcgagga agtgggcgga
    14401 gtccggtttt ggcgccagcc gctgaggctg ccaagcagaa aagccaccgc tgaggagact
    14461 ccggtcactg tcctcgcccc gcctccccct tccctcccct tggggaccac cgggcgccac
    14521 gccgcgaacg gtaagtgccg cggtcgtcgg cgcctccgcc ctccccctag ggccccaatt
    14581 cccagcgggc gcggcgcgcg gcccctcccc ccgccgggcg cgcgcccgct gccccgccct
    14641 tcgtggccgc ccggcgtggg cggtgccacc cctccccccg gcggccccgc gcgcagctcc
    14701 cggctccctc ccccttcgga tgtggcttga gctgtaggcg cggaggctag ctggggatct
    14761 ggtgggctct attggtagac tttagtccct atcagtgata gagaaggtat ctccagttta
    14821 ctccctatca gtgatagaga catccatgga atcaacttca gctttatgag taagaagccc
    14881 tataggatgt ggctttccct atcagtgata gagaactaat gataagacca ctgcacttta
    14941 gtccctatca gtgatagaga tgagatcttc tggtgtacag gagttttctc cctatcagtg
    15001 atagagagtc tgtgtgccat ttgatcccta tcagtgatag agactgtatt tccaggtagg
    15061 cctgtacagt gggaggccta tataagcaga gctcctttag tgaaccctca gatggccaga
    15121 gataagagag aaaagaagag taagaagaaa tataagagcc accacaagtt tgtacaaaaa
    15181 agcaggcttc gccgccatgc agagcaccgt gcccagagcc agggacccac ctgtggccac
    15241 cagcaaaggc gaggaactct ttacgggggt agtaccgatt cttgtggagt tggacggtga
    15301 tgtcaatggt cacaagtttt ctgtccgagg cgaaggggag ggcgatgcga ccaatggtaa
    15361 gcttacattg aagttcattt gtacgacagg caagctgcca gtaccttggc caacgttggt
    15421 gacgacactt acatacggag tacagtgctt cagccgctat cctgatcaca tgaaacgaca
    15481 tgattttttt aaatccgcaa tgcccgaagg gtacgtgcag gaacggacaa tttcctttaa
    15541 agacgacggg acatataaga caagggcgga ggtgaagttt gaaggcgaca ctcttgtgaa
    15601 taggatagaa ctcaaaggca ttgattttaa agaagatggg aacattttgg gtcataagct
    15661 tgagtataac ttcaattccc acaacgtcta cataacagcc gacaaacaaa aaaacggaat
    15721 aaaagcaaac ttcaaaatta ggcataacgt tgaggatggt tccgtccagc tcgccgatca
    15781 ttatcagcag aatacgccta ttggagatgg gccggttctc ctccccgata atcactatct
    15841 gagtacccag agtgtgcttt ctaaagatcc caacgaaaag cgcgatcata tggttctcct
    15901 cgaatttgtc actgcagcag ggattaccca tggtatggac gaattgtata aatgaa
//
