LOCUS       sequence-441837-       15527 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ngn2 mediated cortical neuron induction, dox induction is included.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 15527)
  AUTHORS   Hallacli E, Kayatekin C, Nazeen S, Wang XH, Sheinkopf Z, Sathyakumar
            S, Sarkar S, Jiang X, Dong X, Di Maio R, Wang W, Keeney MT, Felsky 
            D, Sandoe J, Vahdatshoar A, Udeshi ND, Mani DR, Carr SA, Lindquist 
            S, De Jager PL, Bartel DP, Myers CL, Greenamyre JT, Feany MB, 
            Sunyaev SR, Chung CY, Khurana V
  TITLE     The Parkinson's disease protein alpha-synuclein is a modulator of 
            processing bodies and mRNA stability.
  JOURNAL   Cell. 2022 Jun 9;185(12):2035-2056.e33. doi: 
            10.1016/j.cell.2022.05.008.
  PUBMED    35688132
REFERENCE   2  (bases 1 to 15527)
  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"; doi: 
            "10.1016/j.cell.2022.05"; journalName: "Cell"; date: "2022-06-9- 9";
            volume: "185"; issue: "12"; pages: "2035-2056.e33"
COMMENT     Sequence Label: piggyBac-rtTA (4th_Gen)-NGN2-2A-PURO-IRES-SNAP...
FEATURES             Location/Qualifiers
     source          1..15527
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(109..128)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      922..1510
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1411..1430
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1664..1681
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1798..1819
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1834..1839,1840..1857,1858..1864)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1872..1888
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     1877..1899
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1896..1912
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1896..1912
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1931..1951
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1933..1951
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2645..3155
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             3162..4292
                     /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"
     primer_bind     complement(4290..4309)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4334..4382
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     repeat_region   4506..4568
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    6056..6532
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     6314..6333
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    6564..7152
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             6670..6681
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     7079..7099
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7159..7179)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(7189..7734)
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     misc_feature    7746..8298
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    7748..8315
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7908..7927)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     8137..8154
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     protein_bind    8367..8391
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(8393..8992)
                     /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(8476..8496)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     8973..8992
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(8999..9052)
                     /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(9845..9868)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      9868..9877
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(9880..9904)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    10025..10043
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10061..10079
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10108..10126
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10154..10172
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10228..10246
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10264..10282
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        10994..11504
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             11514..11909
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             11919..11972
                     /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             12630..12779
                     /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             12804..12824
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             13254..13610
                     /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             13629..14198
                     /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"
     primer_bind     complement(14221..14238)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    14227..14451
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   15004..15038
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     15219..15238
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        15313..15417
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(15418..15486,15487..15527,1..751)
                     /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"
ORIGIN
        1 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
       61 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
      121 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
      181 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
      241 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
      301 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
      361 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
      421 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
      481 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
      541 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
      601 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
      661 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
      721 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
      781 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
      841 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
      901 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
      961 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     1021 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     1081 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     1141 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     1201 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     1261 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     1321 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     1381 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     1441 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     1501 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     1561 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     1621 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     1681 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     1741 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     1801 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     1861 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     1921 cgccaagcgc gcaattaacc ctcactaaag ggaacaaaag ctggattaat taagggaggt
     1981 ggtccctgca gttacgccaa tgataacccc cgccagaaaa atcttagtag ccttcccttt
     2041 ttgttttccg tgccccaact cggcggattg actcggcccc ttccggaaac acccgaatca
     2101 acttctagtc aaattattgt tcacgccgca atgacccacc cctggcccgc gtctgtggaa
     2161 ctgacccctg gtgtacagga gagttcgctg ctgaaagtgg tcccaaaggg gtactagttt
     2221 ttaagctccc aactccccct cccccagcgt ctggaggatt ccacaccctc gcaccgcagg
     2281 ggcgaggaag tgggcggagt ccggttttgg cgccagccgc tgaggctgcc aagcagaaaa
     2341 gccaccgctg aggagactcc ggtcactgtc ctcgccccgc ctcccccttc cctccccttg
     2401 gggaccaccg ggcgccacgc cgcgaacggt aagtgccgcg gtcgtcggcg cctccgccct
     2461 ccccctaggg ccccaattcc cagcgggcgc ggcgcgcggc ccctcccccc gccgggcgcg
     2521 cgcccgctgc cccgcccttc gtggccgccc ggcgtgggcg gtgccacccc tccccccggc
     2581 ggccccgcgc gcagctcccg gctccctccc ccttcggatg tggcttgagc tgtaggcgcg
     2641 gaggggggtt ggggttgcgc cttttccaag gcagccctgg gtttgcgcag ggacgcggct
     2701 gctctgggcg tggttccggg aaacgcagcg gcgccgaccc tgggtctcgc acattcttca
     2761 cgtccgttcg cagcgtcacc cggatcttcg ccgctaccct tgtgggcccc ccggcgacgc
     2821 ttcctgctcc gcccctaagt cgggaaggtt ccttgcggtt cgcggcgtgc cggacgtgac
     2881 aaacggaagc cgcacgtctc actagtaccc tcgcagacgg acagcgccag ggagcaatgg
     2941 cagcgcgccg accgcgatgg gctgtggcca atagcggctg ctcagcaggg cgcgccgaga
     3001 gcagcggccg ggaaggggcg gtgcgggagg cggggtgtgg ggcggtagtg tgggccctgt
     3061 tcctgcccgc gcggtgttcc gcattctgca agcctccgga gcgcacgtcg gcagtcggct
     3121 ccctcgttga ccgaatcacc gacctctctc cccagaccgc catggcttcg taccccggcc
     3181 atcagcacgc gtctgcgttc gaccaggctg cgcgttctcg cggccatagc aaccgacgta
     3241 cggcgttgcg ccctcgccgg cagcaagaag ccacggaagt ccgcccggag cagaaaatgc
     3301 ccacgctact gcgggtttat atagacggtc cccacgggat ggggaaaacc accaccacgc
     3361 aactgctggt ggccctgggt tcgcgcgacg atatcgtcta cgtacccgag ccgatgactt
     3421 actggcgggt gctgggggct tccgagacaa tcgcgaacat ctacaccaca caacaccgcc
     3481 ttgaccaggg tgagatatcg gccggggacg cggcggtggt aatgacaagc gcccagataa
     3541 caatgggcat gccttatgcc gtgaccgacg ccgttctggc tcctcatatc gggggggagg
     3601 ctgggagctc acatgccccg cccccggccc tcaccctcat cttcgaccgc catcccatcg
     3661 ccgccctcct gtgctacccg gccgcgcgat accttatggg cagcatgacc ccccaggccg
     3721 tgctggcgtt cgtggccctc atcccgccga ccttgcccgg cacaaacatc gtgttggggg
     3781 cccttccgga ggacagacac atcgaccgcc tggccaaacg ccagcgcccc ggcgagcggc
     3841 ttgacctggc tatgctggcc gcgattcgcc gcgtttacgg gctgcttgcc aatacggtgc
     3901 ggtatctgca gggcggcggg tcgtggcggg aggattgggg acagctttcg gggacggccg
     3961 tgccgcccca gggtgccgag ccccagagca acgcgggccc acgaccccat atcggggaca
     4021 cgttatttac cctgtttcgg gcccccgagt tgctggcccc caacggcgac ctgtacaacg
     4081 tgtttgcctg ggccttggac gtcttggcca aacgcctccg tcccatgcac gtctttatcc
     4141 tggattacga ccaatcgccc gccggctgcc gggacgccct gctgcaactt acctccggga
     4201 tgatccagac ccacgtcacc accccaggct ccataccgac gatctgcgac ctggcgcgca
     4261 cgtttgcccg ggagatgggg gaggctaact gaaacacgga aggagacaat accggaagga
     4321 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt
     4381 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     4441 ttggggccaa tacgcccgcg tttcttcctt ttccccagcg gccgcatacg cgtatactag
     4501 attaacccta gaaagataat catattgtga cgtacgttaa agataatcat gcgtaaaatt
     4561 gacgcatgtg ttttatcggt ctgtatatcg aggtttattt attaatttga atagatatta
     4621 agttttatta tatttacact tacatactaa taataaattc aacaaacaat ttatttatgt
     4681 ttatttattt attaaaaaaa aacaaaaact caaaatttct tctataaagt aacaaaactt
     4741 ttaaaaggac agggaaggga gcagtggttc acgcctgtaa tcccagcaat ggcgcgccta
     4801 taatgtttaa acaatattct cactgactcc gtcctggagt tggatgagag ataatggcct
     4861 tacgttgtgc caggggaggg tcgggctgga tttagcaaga tttaccttct ccaaagagcg
     4921 gtgctgcagt ggcacagctg cccacggagg tgggggggtc accgtccctg gaggtgatga
     4981 agaactgtgg ggatgtggca ctgagggaca tggccagtgg gcacggtggg tgggttgggg
     5041 ttggtcttgg ggatcttgga gggcttttcc agccttcatg atttgacgat tgtatgaaca
     5101 tctacatggc aattctccag ctgcctgtcc cagtcctact gacccagctg tatctctcca
     5161 ggcaagctct tccacccctt ctgcttgcat ccagacacca tcaaacatgc aggctcagac
     5221 acagggacca gcagtgtctg tggccttttt gtgctcctct ccatgctggg ttttaacttg
     5281 ctctttgtcc ttctatccta tcttcttatc cttaaggctg ttctgaacgc tgtgacttgg
     5341 agagtgtccc agagccctca acacctgcat gtcccacgtc catgctgtcc tgcacttcct
     5401 tatccccaag atctgcctct ccgtgatgca ctgaattggc aaacatgtgt caccccagac
     5461 caacaatgtc acagcaaact cccccttgat aggacaaggg ggaatggctt tacactgaga
     5521 caggggaggt ttgggttgga tatgaggagg cagtttttcc cccagagggt ggtgacgcac
     5581 tgaacaggtt gcccaaggag gctgtggatg ccccatccct gcaggcattc aaggccaggc
     5641 tggatgtggc tctgggcagc ctgggctgct ggttgatgac cctgcacata gcagggggtt
     5701 ggatctggat gagcactgtg ctcctttgca acccaggccg ttctatgatt ctgtcattct
     5761 aaatctctct ttcagcctaa agctttttcc ccgtatcccc ccaggtgtct gcaggctcaa
     5821 agagcagcga gaagcgttca gaggaaagcg atcccgtgcc accttccccg tgcccgggct
     5881 gtccccgcac gctgccggct cggggatgcg gggggagcgc cggaccggag cggagccccg
     5941 ggcggctcgc tgctgccccc tagcggggga gggacgtaat tacatccctg ggggctttgg
     6001 ggggggggct gtccccgtga gctcagtact atgcatgtta acttaattaa ggtacaagga
     6061 cagggaaggg agcagtggtt cacgcctgta atcccagcaa tttgggaggc caaggtgggt
     6121 agatcacctg agattaggag ttggagacca gcctggccaa tatggtgaaa ccccgtctct
     6181 accaaaaaaa caaaaattag ctgagcctgg tcatgcatgc ctggaatccc aacaactcgg
     6241 gaggctgagg caggagaatc gcttgaaccc aggaggcgga gattgcagtg agccaagatt
     6301 gtgccactgc actccagctt ggttcccaat agaccccgca ggccctacag gttgtcttcc
     6361 caacttgccc cttgctccat accacccccc tccaccccat aatattatag aaggacacct
     6421 agtcagacaa aatgatgcaa cttaatttta ttaggacaag gctggtgggc actggagtgg
     6481 caacttccag ggccaggaga ggcactgggg aggggtcaca gggatgccac ccgtagatct
     6541 ctcgagcagc gctcggtatc gatgcgggga ggcggcccaa agggagatcc gactcgtctg
     6601 agggcgaagg cgaagacgcg gaagaggccg cagagccggc agcaggccgc gggaaggaag
     6661 gtccgctgga ttgagggccg aagggacgta gcagaaggac gtcccgcgca gaatccaggt
     6721 ggcaacatag gcgagcagcc aaggaaagga cgatgatttc cccgacaaca ccacggaatt
     6781 gtcagtgccc aacagccgag cccctgtcca gcagcgggca aggcaggcgg cgatgagttc
     6841 cgccgtggca atagggaggg ggaaagcgaa agttccggaa aggagctgac aggtggtggc
     6901 aatgccccaa ccagtggggg ttgcgtcagc aaacacagtg cacaccacgc cacgttgcct
     6961 gacaacgggc cacaactcct cataaagaga cagcaaccag gatttataca aggaggagaa
     7021 aatgaaagcc atacgggaag caatagcatg atacaaaggc attaaagcag cgtatccaca
     7081 tagcgtaaaa ggagcaacat agttaagaat accagtcaat ctttcacaaa ttttgtaatc
     7141 cagaggttga ttccgctaca ccttccgctt cttctttggg ctcccgccac ccagcccagg
     7201 cttgcccagt ctgtggccct cgtgggccag cagccactct ttcacggcga gcccgccctc
     7261 gtagcccccc acgtccaggt cgccctgcac cacccggtgg caggggatca gaatgggcac
     7321 gggatttccg ctcagggcgg ttttcacggc ggcggtggcg gcgggattgc cggccagggc
     7381 ggccaggtgg ctgtagctga tgacctctcc gaacttcacc actttcagca gtttccacag
     7441 cacctggcgg gtaaagctct cctgctggaa cactgggtgg tgcagggctg gcacagggaa
     7501 ctcctcgatg gcctcaggct ggtgaaagta ggcgttgagc caggcggtgg cctgcatcag
     7561 tggctctggt ccgcccagca cggcggctgg ggcaggcact tccacggcgt cggcggcaga
     7621 tgttcctttg cccaggaaga tgatacggtg caggccctgt tcgcacccag acagttccag
     7681 cttgcccaga gggctatcca gggtggtgcg cttcatttcg cagtctttgt ccatttgtgt
     7741 ggccatatta tcatcgtgtt tttcaaagga aaaccacgtc cccgtggttc ggggggccta
     7801 gacgtttttt taacctcgac taaacacatg taaagcatgt gcaccgaggc cccagatcag
     7861 atcccataca atggggtacc ttctgggcat ccttcagccc cttgttgaat acgcttgagg
     7921 agagccattt gactctttcc acaactatcc aactcacaac gtggcactgg ggttgtgccg
     7981 cctttgcagg tgtatcttat acacgtggct tttggccgca gaggcacctg tcgccaggtg
     8041 gggggttccg ctgcctgcaa agggtcgcta cagacgttgt ttgtcttcaa gaagcttcca
     8101 gaggaactgc ttccttcacg acattcaaca gaccttgcat tcctttggcg agaggggaaa
     8161 gacccctagg aatgctcgtc aagaagacag ggccaggttt ccgggccctc acattgccaa
     8221 aagacggcaa tatggtggaa aataacatat agacaaacgc acaccggcct tattccaagc
     8281 ggcttcggcc agtaacgtta gggggggggg agggatctct aatcgccatc ttcgtacctc
     8341 tctaatcgcc atcttccagc agatcaacca ctttgtacaa gaaagctggg tttcaggcac
     8401 cgggcttgcg ggtcatgcac caggtgcgcg gtccttcggg cacctcgacg tcggcggtga
     8461 cggtgaagcc gagccgctcg tagaagggga ggttgcgggg cgcggaggtc tccaggaagg
     8521 cgggcacccc ggcgcgctcg gccgcctcca ctccggggag cacgacggcg ctgcccagac
     8581 ccttgccctg gtggtcgggc gagacgccga cggtggccag gaaccacgcg ggctccttgg
     8641 gccggtgcgg cgccaggagg ccttccatct gttgctgcgc ggccagccgg gaaccgctca
     8701 actcggccat gcgcgggccg atctcggcga acaccgcccc cgcttcgacg ctctccggcg
     8761 tggtccagac cgccaccgcg gcgccgtcgt ccgcgaccca caccttgccg atgtcgagcc
     8821 cgacgcgcgt gaggaagagt tcttgcagct cggtgacccg ctcgatgtgg cggtccggat
     8881 cgacggtgtg gcgcgtggcg gggtagtcgg cgaacgcggc ggcgagggtg cgtacggccc
     8941 tggggacgtc gtcgcgggtg gcgaggcgca ccgtgggctt gtactcggtc atctcgagtg
     9001 ggccgggatt ttcctccacg tccccgcatg ttagaagact tcccctgccc tctctagaga
     9061 tacagtccct ggcgaggggc aggtgaggcg cataacgatg cttctccgga ggtgggggct
     9121 gccagtagtc cacgtctgac ccggggctag cgggcgataa agtgcagctg tatggggacg
     9181 tggagttgga ggatgacgcc gggctgttgg tgcagctcca ggaggaaggt ggagaagggc
     9241 tgtccccgct ggcgccgagc gcagctcccg ggctcaggag caccgcctcc gtgaagagcg
     9301 ccccctggag gccaccggcg ccggcgcagt ggtccgccag gcgcagagtc tcggtgagcg
     9361 cccagatgta attgtgggcg aagcgcagcg tctcgatctt cgtgagcttg gcatcctcgg
     9421 ggaaggtggg cagcacctcg cgcagcgcgt ccagcgcggc gtttaggttg tgcatgcggt
     9481 tgcgctcgcg gttgttggcc ttgagcctgc gggtcttctt gatgcgctgc accgtctccg
     9541 ccgtcttggc acctcgggag acggcccgtg agcgcgacgg gcgacgcttg cactcgtgca
     9601 tcaggcccag cagccgccct ggccggcaac ccccggcacc cgacgccgga ctgccctgca
     9661 ccccctgccc ggcttccgct ccacgctgcc cacgcgcgga gcccggccgg cgcagctcct
     9721 cgtcctcctc ctcgtccgcg ctggaggaca tcggggtcag ggtcgccgag gccggggaag
     9781 ccgagcccag cagcatcagt acctcctctt cctccttcaa ctccagagtc tcagatttga
     9841 cgaacttgtc atcgtcgtcc ttgtagtcca tggtggctaa gcctgctttt ttgtacaaac
     9901 ttgtggtggc tcttatattt cttcttactc ttcttttctc tcttatctct ggccatctga
     9961 gggttcacta aaggagctct gcttatatag gcctcccact gtacaggcct acctggaaat
    10021 acagtctcta tcactgatag ggatcaaatg gcacacagac tctctatcac tgatagggag
    10081 aaaactcctg tacaccagaa gatctcatct ctatcactga tagggactaa agtgcagtgg
    10141 tcttatcatt agttctctat cactgatagg gaaagccaca tcctataggg cttcttactc
    10201 ataaagctga agttgattcc atggatgtct ctatcactga tagggagtaa actggagata
    10261 ccttctctat cactgatagg gactaaagtc taccaataga gcccaccaga tccccagcta
    10321 gcctccgcgc ctacagctca agccacatcc gaagggggag ggagccggga gctgcgcgcg
    10381 gggccgccgg ggggaggggt ggcaccgccc acgccgggcg gccacgaagg gcggggcagc
    10441 gggcgcgcgc ccggcggggg gaggggccgc gcgccgcgcc cgctgggaat tggggcccta
    10501 gggggagggc ggaggcgccg acgaccgcgg cacttaccgt tcgcggcgtg gcgcccggtg
    10561 gtccccaagg ggagggaagg gggaggcggg gcgaggacag tgaccggagt ctcctcagcg
    10621 gtggcttttc tgcttggcag cctcagcggc tggcgccaaa accggactcc gcccacttcc
    10681 tcgcccctgc ggtgcgaggg tgtggaatcc tccagacgct gggggagggg gagttgggag
    10741 cttaaaaact agtacccctt tgggaccact ttcagcagcg aactctcctg tacaccaggg
    10801 gtcagttcca cagacgcggg ccaggggtgg gtcattgcgg cgtgaacaat aatttgacta
    10861 gaagttgatt cgggtgtttc cggaaggggc cgagtcaatc cgccgagttg gggcacggaa
    10921 aacaaaaagg gaaggctact aagatttttc tggcgggggt tatcattggc gtaactgcag
    10981 ggaccacctc cccggggttg gggttgcgcc ttttccaagg cagccctggg tttgcgcagg
    11041 gacgcggctg ctctgggcgt ggttccggga aacgcagcgg cgccgaccct gggtctcgca
    11101 cattcttcac gtccgttcgc agcgtcaccc ggatcttcgc cgctaccctt gtgggccccc
    11161 cggcgacgct tcctgctccg cccctaagtc gggaaggttc cttgcggttc gcggcgtgcc
    11221 ggacgtgaca aacggaagcc gcacgtctca ctagtaccct cgcagacgga cagcgccagg
    11281 gagcaatggc agcgcgccga ccgcgatggg ctgtggccaa tagcggctgc tcagcagggc
    11341 gcgccgagag cagcggccgg gaaggggcgg tgcgggaggc ggggtgtggg gcggtagtgt
    11401 gggccctgtt cctgcccgcg cggtgttccg cattctgcaa gcctccggag cgcacgtcgg
    11461 cagtcggctc cctcgttgac cgaatcaccg acctctctcc ccagcaattc accatggcca
    11521 agcctttgtc tcaagaagaa tccaccctca ttgaaagagc aacggctaca atcaacagca
    11581 tccccatctc tgaagactac agcgtcgcca gcgcagctct ctctagcgac ggccgcatct
    11641 tcactggtgt caatgtatat cattttactg ggggaccttg tgcagaactc gtggtgctgg
    11701 gcactgctgc tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga
    11761 acaggggcat cttgagcccc tgcggacggt gccgacaggt gcttctcgat ctgcatcctg
    11821 ggatcaaagc catagtgaag gacagtgatg gacagccgac ggcagttggg attcgtgaat
    11881 tgctgccctc tggttatgtg tgggagggcg gatccggcga aggtagaggt tctctcctca
    11941 cttgtggtga tgttgaagaa aaccctggtc caggtggttc tatgtctaga ctggacaaga
    12001 gcaaagtcat aaactctgct ctggaattac tcaatggagt cggtatcgaa ggcctgacga
    12061 caaggaaact cgctcaaaag ctgggagttg agcagcctac cctgtactgg cacgtgaaga
    12121 acaagcgggc cctgctcgat gccctgccaa tcgagatgct ggacaggcat catacccact
    12181 cctgccccct ggaaggcgag tcatggcaag actttctgcg gaacaacgcc aagtcatacc
    12241 gctgtgctct cctctcacat cgcgacgggg ctaaagtgca tctcggcacc cgcccaacag
    12301 agaaacagta cgaaaccctg gaaaatcagc tcgcgttcct gtgtcagcaa ggcttctccc
    12361 tggagaacgc actgtacgct ctgtccgccg tgggccactt tacactgggc tgcgtattgg
    12421 aggaacagga gcatcaagta gcaaaagagg aaagagagac acctaccacc gattctatgc
    12481 ccccacttct gaaacaagca attgagctgt tcgaccggca gggagccgaa cctgccttcc
    12541 ttttcggcct ggaactaatc atatgtggcc tggagaaaca gctaaagtgc gaaagcggcg
    12601 ggccggaggc cagcggttcc ggacgggctg acgcattgga cgattttgat ctggatatgc
    12661 tgggaagtga cgccctcgat gattttgacc ttgacatgct tggttcggat gcccttgatg
    12721 actttgacct cgacatgctc ggcagtgacg cccttgatga tttcgacctg gacatgctga
    12781 ttaactctag aagttccgga tctccgaaaa agaaacgcaa agttggtagc cagtacctgc
    12841 ccgacaccga cgaccggcac cggatcgagg aaaagcggaa gcggacctac gagacattca
    12901 agagcatcat gaagaagtcc cccttcagcg gccccaccga ccctagacct ccacctagaa
    12961 gaatcgccgt gcccagcaga tccagcgcca gcgtgccaaa acctgccccc cagccttacc
    13021 ccttcaccag cagcctgagc accatcaact acgacgagtt ccctaccatg gtgttcccca
    13081 gcggccagat ctctcaggcc tctgctctgg ctccagcccc tcctcaggtg ctgcctcagg
    13141 ctcctgctcc tgcaccagct ccagccatgg tgtctgcact ggctcaggca ccagcacccg
    13201 tgcctgtgct ggctcctgga cctccacagg ctgtggctcc accagcccct aaacctacac
    13261 aggccggcga gggcacactg tctgaagctc tgctgcagct gcagttcgac gacgaggatc
    13321 tgggagccct gctgggaaac agcaccgatc ctgccgtgtt caccgacctg gccagcgtgg
    13381 acaacagcga gttccagcag ctgctgaacc agggcatccc tgtggcccct cacaccaccg
    13441 agcccatgct gatggaatac cccgaggcca tcacccggct cgtgacaggc gctcagaggc
    13501 ctcctgatcc agctcctgcc cctctgggag caccaggcct gcctaatgga ctgctgtctg
    13561 gcgacgagga cttcagctct atcgccgata tggatttctc agccttgctg ggctctggca
    13621 gcggcagccg ggattccagg gaagggatgt ttttgccgaa gcctgaggcc ggctccgcta
    13681 ttagtgacgt gtttgagggc cgcgaggtgt gccagccaaa acgaatccgg ccatttcatc
    13741 ctccaggaag tccatgggcc aaccgcccac tccccgccag cctcgcacca acaccaaccg
    13801 gtccagtaca tgagccagtc gggtcactga ccccggcacc agtccctcag ccactggatc
    13861 cagcgcccgc agtgactccc gaggccagtc acctgttgga ggatcccgat gaagagacga
    13921 gccaggctgt caaagccctt cgggagatgg ccgatactgt gattccccag aaggaagagg
    13981 ctgcaatctg tggccaaatg gacctttccc atccgccccc aaggggccat ctggatgagc
    14041 tgacaaccac acttgagtcc atgaccgagg atctgaacct ggactcaccc ctgaccccgg
    14101 aattgaacga gattctggat accttcctga acgacgagtg cctcttgcat gccatgcata
    14161 tcagcacagg actgtccatc ttcgacacat ctctgttttg agtttaaacc cgctgatcag
    14221 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
    14281 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
    14341 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
    14401 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg ggtctcgaca
    14461 ttcattctag ttgatcgtct cgaactcctg acccctcaca aagtgctggg attacaagcg
    14521 tgagccaccg cgcccggcca gaacttttga aaggaaattt acaaaggtag ttattggata
    14581 aaagcgtgat agagtagtta gcaccaggtg gcgctaaagg actgaaattg gaaacggcta
    14641 cggtatttgg tcctcttacc tgagggccgg ggaagggcga tgctcctggt ttaatggctg
    14701 tgttacagga aggaaggaca agcatttgtg atatctataa caagaaaata tatatataat
    14761 aagttatcac gtaagtagaa catgaaataa caatataatt atcgtatgag ttaaatctta
    14821 aaagtcacgt aaaagataat catgcgtcat tttgactcac gcggtcgtta tagttcaaaa
    14881 tcagtgacac ttaccgcatt gacaagcacg cctcacggga gctccaagcg gcgactgaga
    14941 tgtcctaaat gcacagcgac ggattcgcgc tatttagaaa gagagagcaa tatttcaaga
    15001 atgcatgcgt caattttacg cagactatct ttctagggtt aatctagagc tcagttagcc
    15061 tcccccatct ccccgaagca gcgcaaaacg cctaacccta agcagattct tcatgcaatt
    15121 gtcggtcaag ccttgccttg ttgtagctta aattttgctc gcgcactact cagcgacctc
    15181 caacacacaa gcagggagca gatactggct taactatgcg gcatcagagc agattgtact
    15241 gagagtgcac cataggggat cgggagatct cccgatccgt cgacgtcagg tggcactttt
    15301 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
    15361 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
    15421 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
    15481 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatca
//
