LOCUS       sequence-417159-       14375 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ngn2 mediated cortical neuron induction, SNCA(Avi tagged) 
            expression, Synapsin promoter BirA ligase expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14375)
  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 14375)
  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)_SNCA-Avi_IRES_NGN2-2A-...
FEATURES             Location/Qualifiers
     source          1..14375
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   132..166
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     polyA_signal    719..943
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     932..949
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(972..1541)
                     /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(1560..1916)
                     /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(2346..2366)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(2391..2540)
                     /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(3198..3251)
                     /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(3261..3656)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     promoter        complement(3666..4176)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3729..3747)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     protein_bind    4888..4906
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4924..4942
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4998..5016
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5044..5062
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5091..5109
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5127..5145
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5266..5290
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      5294..5303
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5726..5770
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     protein_bind    complement(5775..5799)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    5851..6418
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5868..6420
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6012..6029)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6239..6258
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             6432..6455
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             7248..7301
                     /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             7308..7907
                     /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(7308..7327)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7804..7824
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    7908..8496
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7961..7981)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8379..8390)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    8511..8987
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(8710..8729)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     promoter        9035..9482
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             join(9496..9846,9847..9849,9850..10458)
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             9496..10458
                     /codon_start=1
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     polyA_signal    10462..10510
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     repeat_region   11975..12037
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(12067..12087)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(12067..12085)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(12106..12122)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(12106..12122)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(12119..12141)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    12130..12146
                     /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(12154..12160,12161..12178,12179..12184))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12199..12220
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(12337..12354)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(12508..13096)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(12588..12607)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(13267..14058,14059..14127))
                     /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     13890..13909
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(14128..14232)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(14307..14326)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
       61 gaatctgctt agggttaggc gttttgcgct gcttcgggga gatgggggag gctaactgag
      121 ctctagatta accctagaaa gatagtctgc gtaaaattga cgcatgcatt cttgaaatat
      181 tgctctctct ttctaaatag cgcgaatccg tcgctgtgca tttaggacat ctcagtcgcc
      241 gcttggagct cccgtgaggc gtgcttgtca atgcggtaag tgtcactgat tttgaactat
      301 aacgaccgcg tgagtcaaaa tgacgcatga ttatctttta cgtgactttt aagatttaac
      361 tcatacgata attatattgt tatttcatgt tctacttacg tgataactta ttatatatat
      421 attttcttgt tatagatatc acaaatgctt gtccttcctt cctgtaacac agccattaaa
      481 ccaggagcat cgcccttccc cggccctcag gtaagaggac caaataccgt agccgtttcc
      541 aatttcagtc ctttagcgcc acctggtgct aactactcta tcacgctttt atccaataac
      601 tacctttgta aatttccttt caaaagttct ggccgggcgc ggtggctcac gcttgtaatc
      661 ccagcacttt gtgaggggtc aggagttcga gacgatcaac tagaatgaat gtcgagaccc
      721 atagagccca ccgcatcccc agcatgcctg ctattgtctt cccaatcctc ccccttgctg
      781 tcctgcccca ccccaccccc cagaatagaa tgacacctac tcagacaatg cgatgcaatt
      841 tcctcatttt attaggaaag gacagtggga gtggcacctt ccagggtcaa ggaaggcacg
      901 ggggaggggc aaacaacaga tggctggcaa ctagaaggca cagtcgaggc tgatcagcgg
      961 gtttaaactc aaaacagaga tgtgtcgaag atggacagtc ctgtgctgat atgcatggca
     1021 tgcaagaggc actcgtcgtt caggaaggta tccagaatct cgttcaattc cggggtcagg
     1081 ggtgagtcca ggttcagatc ctcggtcatg gactcaagtg tggttgtcag ctcatccaga
     1141 tggccccttg ggggcggatg ggaaaggtcc atttggccac agattgcagc ctcttccttc
     1201 tggggaatca cagtatcggc catctcccga agggctttga cagcctggct cgtctcttca
     1261 tcgggatcct ccaacaggtg actggcctcg ggagtcactg cgggcgctgg atccagtggc
     1321 tgagggactg gtgccggggt cagtgacccg actggctcat gtactggacc ggttggtgtt
     1381 ggtgcgaggc tggcggggag tgggcggttg gcccatggac ttcctggagg atgaaatggc
     1441 cggattcgtt ttggctggca cacctcgcgg ccctcaaaca cgtcactaat agcggagccg
     1501 gcctcaggct tcggcaaaaa catcccttcc ctggaatccc ggctgccgct gccagagccc
     1561 agcaaggctg agaaatccat atcggcgata gagctgaagt cctcgtcgcc agacagcagt
     1621 ccattaggca ggcctggtgc tcccagaggg gcaggagctg gatcaggagg cctctgagcg
     1681 cctgtcacga gccgggtgat ggcctcgggg tattccatca gcatgggctc ggtggtgtga
     1741 ggggccacag ggatgccctg gttcagcagc tgctggaact cgctgttgtc cacgctggcc
     1801 aggtcggtga acacggcagg atcggtgctg tttcccagca gggctcccag atcctcgtcg
     1861 tcgaactgca gctgcagcag agcttcagac agtgtgccct cgccggcctg tgtaggttta
     1921 ggggctggtg gagccacagc ctgtggaggt ccaggagcca gcacaggcac gggtgctggt
     1981 gcctgagcca gtgcagacac catggctgga gctggtgcag gagcaggagc ctgaggcagc
     2041 acctgaggag gggctggagc cagagcagag gcctgagaga tctggccgct ggggaacacc
     2101 atggtaggga actcgtcgta gttgatggtg ctcaggctgc tggtgaaggg gtaaggctgg
     2161 ggggcaggtt ttggcacgct ggcgctggat ctgctgggca cggcgattct tctaggtgga
     2221 ggtctagggt cggtggggcc gctgaagggg gacttcttca tgatgctctt gaatgtctcg
     2281 taggtccgct tccgcttttc ctcgatccgg tgccggtcgt cggtgtcggg caggtactgg
     2341 ctaccaactt tgcgtttctt tttcggagat ccggaacttc tagagttaat cagcatgtcc
     2401 aggtcgaaat catcaagggc gtcactgccg agcatgtcga ggtcaaagtc atcaagggca
     2461 tccgaaccaa gcatgtcaag gtcaaaatca tcgagggcgt cacttcccag catatccaga
     2521 tcaaaatcgt ccaatgcgtc agcccgtccg gaaccgctgg cctccggccc gccgctttcg
     2581 cactttagct gtttctccag gccacatatg attagttcca ggccgaaaag gaaggcaggt
     2641 tcggctccct gccggtcgaa cagctcaatt gcttgtttca gaagtggggg catagaatcg
     2701 gtggtaggtg tctctctttc ctcttttgct acttgatgct cctgttcctc caatacgcag
     2761 cccagtgtaa agtggcccac ggcggacaga gcgtacagtg cgttctccag ggagaagcct
     2821 tgctgacaca ggaacgcgag ctgattttcc agggtttcgt actgtttctc tgttgggcgg
     2881 gtgccgagat gcactttagc cccgtcgcga tgtgagagga gagcacagcg gtatgacttg
     2941 gcgttgttcc gcagaaagtc ttgccatgac tcgccttcca gggggcagga gtgggtatga
     3001 tgcctgtcca gcatctcgat tggcagggca tcgagcaggg cccgcttgtt cttcacgtgc
     3061 cagtacaggg taggctgctc aactcccagc ttttgagcga gtttccttgt cgtcaggcct
     3121 tcgataccga ctccattgag taattccaga gcagagttta tgactttgct cttgtccagt
     3181 ctagacatag aaccacctgg accagggttt tcttcaacat caccacaagt gaggagagaa
     3241 cctctacctt cgccggatcc gccctcccac acataaccag agggcagcaa ttcacgaatc
     3301 ccaactgccg tcggctgtcc atcactgtcc ttcactatgg ctttgatccc aggatgcaga
     3361 tcgagaagca cctgtcggca ccgtccgcag gggctcaaga tgcccctgtt ctcatttccg
     3421 atcgcgacga tacaagtcag gttgccagct gccgcagcag cagcagtgcc cagcaccacg
     3481 agttctgcac aaggtccccc agtaaaatga tatacattga caccagtgaa gatgcggccg
     3541 tcgctagaga gagctgcgct ggcgacgctg tagtcttcag agatggggat gctgttgatt
     3601 gtagccgttg ctctttcaat gagggtggat tcttcttgag acaaaggctt ggccatggtg
     3661 aattgctggg gagagaggtc ggtgattcgg tcaacgaggg agccgactgc cgacgtgcgc
     3721 tccggaggct tgcagaatgc ggaacaccgc gcgggcagga acagggccca cactaccgcc
     3781 ccacaccccg cctcccgcac cgccccttcc cggccgctgc tctcggcgcg ccctgctgag
     3841 cagccgctat tggccacagc ccatcgcggt cggcgcgctg ccattgctcc ctggcgctgt
     3901 ccgtctgcga gggtactagt gagacgtgcg gcttccgttt gtcacgtccg gcacgccgcg
     3961 aaccgcaagg aaccttcccg acttaggggc ggagcaggaa gcgtcgccgg ggggcccaca
     4021 agggtagcgg cgaagatccg ggtgacgctg cgaacggacg tgaagaatgt gcgagaccca
     4081 gggtcggcgc cgctgcgttt cccggaacca cgcccagagc agccgcgtcc ctgcgcaaac
     4141 ccagggctgc cttggaaaag gcgcaacccc aaccccgggg aggtggtccc tgcagttacg
     4201 ccaatgataa cccccgccag aaaaatctta gtagccttcc ctttttgttt tccgtgcccc
     4261 aactcggcgg attgactcgg ccccttccgg aaacacccga atcaacttct agtcaaatta
     4321 ttgttcacgc cgcaatgacc cacccctggc ccgcgtctgt ggaactgacc cctggtgtac
     4381 aggagagttc gctgctgaaa gtggtcccaa aggggtacta gtttttaagc tcccaactcc
     4441 ccctccccca gcgtctggag gattccacac cctcgcaccg caggggcgag gaagtgggcg
     4501 gagtccggtt ttggcgccag ccgctgaggc tgccaagcag aaaagccacc gctgaggaga
     4561 ctccggtcac tgtcctcgcc ccgcctcccc cttccctccc cttggggacc accgggcgcc
     4621 acgccgcgaa cggtaagtgc cgcggtcgtc ggcgcctccg ccctccccct agggccccaa
     4681 ttcccagcgg gcgcggcgcg cggcccctcc ccccgccggg cgcgcgcccg ctgccccgcc
     4741 cttcgtggcc gcccggcgtg ggcggtgcca cccctccccc cggcggcccc gcgcgcagct
     4801 cccggctccc tcccccttcg gatgtggctt gagctgtagg cgcggaggct agctggggat
     4861 ctggtgggct ctattggtag actttagtcc ctatcagtga tagagaaggt atctccagtt
     4921 tactccctat cagtgataga gacatccatg gaatcaactt cagctttatg agtaagaagc
     4981 cctataggat gtggctttcc ctatcagtga tagagaacta atgataagac cactgcactt
     5041 tagtccctat cagtgataga gatgagatct tctggtgtac aggagttttc tccctatcag
     5101 tgatagagag tctgtgtgcc atttgatccc tatcagtgat agagactgta tttccaggta
     5161 ggcctgtaca gtgggaggcc tatataagca gagctccttt agtgaaccct cagatggcca
     5221 gagataagag agaaaagaag agtaagaaga aatataagag ccaccacaag tttgtacaaa
     5281 aaagcaggct gccgccacca tggatgtatt catgaaagga ctttcaaagg ccaaggaggg
     5341 agttgtggct gctgctgaga aaaccaaaca gggtgtggca gaagcagcag gaaagacaaa
     5401 agagggtgtt ctctatgtag gctccaaaac caaggaggga gtggtgcatg gtgtggcaac
     5461 agtggctgag aagaccaaag agcaagtgac aaatgttgga ggagcagtgg tgacgggtgt
     5521 gacagcagta gcccagaaga cagtggaggg agcagggagc attgcagcag ccactggctt
     5581 tgtcaaaaag gaccagttgg gcaagaatga agaaggagcc ccacaggaag gaattctgga
     5641 agatatgcct gtggatcctg acaatgaggc ttatgaaatg ccttctgagg aagggtatca
     5701 agactacgaa cctgaagccc tcgagggtct gaacgacatc ttcgaggctc agaaaatcga
     5761 atggcacgaa taaaacccag ctttcttgta caaagtggtt gatctgctgg aagatggcga
     5821 ttagagaggt acgaagatgg cgattagaga tccctccccc ccccctaacg ttactggccg
     5881 aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc
     5941 gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag
     6001 gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt
     6061 tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa
     6121 ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc
     6181 aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg
     6241 gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat
     6301 gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa
     6361 acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataata
     6421 tggccacaat ggactacaag gacgacgatg acaagttcgt caaatctgag actctggagt
     6481 tgaaggagga agaggaggta ctgatgctgc tgggctcggc ttccccggcc tcggcgaccc
     6541 tgaccccgat gtcctccagc gcggacgagg aggaggacga ggagctgcgc cggccgggct
     6601 ccgcgcgtgg gcagcgtgga gcggaagccg ggcagggggt gcagggcagt ccggcgtcgg
     6661 gtgccggggg ttgccggcca gggcggctgc tgggcctgat gcacgagtgc aagcgtcgcc
     6721 cgtcgcgctc acgggccgtc tcccgaggtg ccaagacggc ggagacggtg cagcgcatca
     6781 agaagacccg caggctcaag gccaacaacc gcgagcgcaa ccgcatgcac aacctaaacg
     6841 ccgcgctgga cgcgctgcgc gaggtgctgc ccaccttccc cgaggatgcc aagctcacga
     6901 agatcgagac gctgcgcttc gcccacaatt acatctgggc gctcaccgag actctgcgcc
     6961 tggcggacca ctgcgccggc gccggtggcc tccagggggc gctcttcacg gaggcggtgc
     7021 tcctgagccc gggagctgcg ctcggcgcca gcggggacag cccttctcca ccttcctcct
     7081 ggagctgcac caacagcccg gcgtcatcct ccaactccac gtccccatac agctgcactt
     7141 tatcgcccgc tagccccggg tcagacgtgg actactggca gcccccacct ccggagaagc
     7201 atcgttatgc gcctcacctg cccctcgcca gggactgtat ctctagagag ggcaggggaa
     7261 gtcttctaac atgcggggac gtggaggaaa atcccggccc actcgagatg accgagtaca
     7321 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg
     7381 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc
     7441 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
     7501 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
     7561 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
     7621 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
     7681 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
     7741 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
     7801 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
     7861 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaaat caacctctgg
     7921 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     7981 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     8041 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     8101 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     8161 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     8221 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     8281 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca
     8341 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     8401 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     8461 agacgagtcg gatctccctt tgggccgcct ccccgcctcg agagatctac gggtggcatc
     8521 cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca
     8581 gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata
     8641 ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg
     8701 cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa
     8761 tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc
     8821 aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca
     8881 tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca
     8941 aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttgta ccttaattaa
     9001 gttaactgct agactgcaga gggccctgcg tatgagtgca agtgggtttt aggaccagga
     9061 tgaggcgggg tgggggtgcc tacctgacga ccgaccccga cccactggac aagcacccaa
     9121 cccccattcc ccaaattgcg catcccctat cagagagggg gaggggaaac aggatgcggc
     9181 gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt gccttcgccc ccgcctggcg
     9241 gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac gtcactcgcc ggtcccccgc
     9301 aaactcccct tcccggccac cttggtcgcg tccgcgccgc cgccggccca gccggaccgc
     9361 accacgcgag gcgcgagata ggggggcacg ggcgcgacca tctgcgctgc ggcgccggcg
     9421 actcagcgct gcctcagtct gcggtgggca gcggaggagt cgtgtcgtgc ctgagagcgc
     9481 agtcgagccg ccaccatgaa ggacaacacc gtgcccctga agctgatcgc cctgctggcc
     9541 aacggcgagt tccactctgg cgagcagctg ggagagaccc tgggaatgag cagagccgcc
     9601 atcaacaagc acatccagac actgagagac tggggagtgg acgtgttcac cgtgcctggc
     9661 aagggctaca gcctgcctga gcctatccag ctgctgaacg ccaagcagat cctgggacag
     9721 ctggatggcg gaagcgtggc cgtgctgcct gtgatcgact ccaccaatca gtacctgctg
     9781 gacagaatcg gagagctgaa gtccggcgac gcctgcatcg ccgagtacca gcaggctggc
     9841 agaggaagaa gaggacggaa gtggttcagc ccattcggag ccaacctgta cctgtccatg
     9901 ttctggagac tggagcaggg acctgctgct gccatcggac tgagtctggt gatcggaatc
     9961 gtgatggccg aggtgctgag aaagctggga gccgacaagg tgagagtgaa gtggcctaat
    10021 gacctgtacc tccaggaccg caagctggct ggcatcctgg tggagctgac aggcaagaca
    10081 ggcgatgccg ctcagatcgt gatcggagcc ggaatcaaca tggccatgag aagagtggag
    10141 gagagcgtgg tgaaccaggg ctggatcacc ctgcaggagg ctggcatcaa cctggaccgg
    10201 aacaccctgg ccgccatgct gatcagagag ctgagagccg ctctggagct gttcgagcag
    10261 gagggactgg ctccttacct gagcagatgg gagaagctgg acaacttcat caacagacct
    10321 gtgaagctga tcatcggcga caaggaaatc ttcggcatct ccagaggaat cgacaagcag
    10381 ggagctctgc tgctggagca ggacggaatc atcaagccct ggatgggcgg agaaatctcc
    10441 ctgagaagcg cagagaagta acggcaataa aaagacagaa taaaacgcac gggtgttggg
    10501 tcgtttgttc gcggccgcga gctcacgggg acagcccccc ccccaaagcc cccagggatg
    10561 taattacgtc cctcccccgc tagggggcag cagcgagccg cccggggctc cgctccggtc
    10621 cggcgctccc cccgcatccc cgagccggca gcgtgcgggg acagcccggg cacggggaag
    10681 gtggcacggg atcgctttcc tctgaacgct tctcgctgct ctttgagcct gcagacacct
    10741 ggggggatac ggggaaaaag ctttaggctg aaagagagat ttagaatgac agaatcatag
    10801 aacggcctgg gttgcaaagg agcacagtgc tcatccagat ccaaccccct gctatgtgca
    10861 gggtcatcaa ccagcagccc aggctgccca gagccacatc cagcctggcc ttgaatgcct
    10921 gcagggatgg ggcatccaca gcctccttgg gcaacctgtt cagtgcgtca ccaccctctg
    10981 ggggaaaaac tgcctcctca tatccaaccc aaacctcccc tgtctcagtg taaagccatt
    11041 cccccttgtc ctatcaaggg ggagtttgct gtgacattgt tggtctgggg tgacacatgt
    11101 ttgccaattc agtgcatcac ggagaggcag atcttgggga taaggaagtg caggacagca
    11161 tggacgtggg acatgcaggt gttgagggct ctgggacact ctccaagtca cagcgttcag
    11221 aacagcctta aggataagaa gataggatag aaggacaaag agcaagttaa aacccagcat
    11281 ggagaggagc acaaaaaggc cacagacact gctggtccct gtgtctgagc ctgcatgttt
    11341 gatggtgtct ggatgcaagc agaaggggtg gaagagcttg cctggagaga tacagctggg
    11401 tcagtaggac tgggacaggc agctggagaa ttgccatgta gatgttcata caatcgtcaa
    11461 atcatgaagg ctggaaaagc cctccaagat ccccaagacc aaccccaacc cacccaccgt
    11521 gcccactggc catgtccctc agtgccacat ccccacagtt cttcatcacc tccagggacg
    11581 gtgacccccc cacctccgtg ggcagctgtg ccactgcagc accgctcttt ggagaaggta
    11641 aatcttgcta aatccagccc gaccctcccc tggcacaacg taaggccatt atctctcatc
    11701 caactccagg acggagtcag tgagaatatt gtttaaacat tataggcgcg ccattgctgg
    11761 gattacaggc gtgaaccact gctcccttcc ctgtcctttt aaaagttttg ttactttata
    11821 gaagaaattt tgagtttttg ttttttttta ataaataaat aaacataaat aaattgtttg
    11881 ttgaatttat tattagtatg taagtgtaaa tataataaaa cttaatatct attcaaatta
    11941 ataaataaac ctcgatatac agaccgataa aacacatgcg tcaattttac gcatgattat
    12001 ctttaacgta cgtcacaata tgattatctt tctagggtta agcgatcgct aatccagctt
    12061 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
    12121 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
    12181 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
    12241 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
    12301 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
    12361 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
    12421 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
    12481 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
    12541 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
    12601 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
    12661 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
    12721 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
    12781 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
    12841 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
    12901 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
    12961 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
    13021 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
    13081 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
    13141 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
    13201 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
    13261 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
    13321 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
    13381 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
    13441 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
    13501 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
    13561 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
    13621 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
    13681 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
    13741 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
    13801 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
    13861 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
    13921 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
    13981 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
    14041 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
    14101 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
    14161 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
    14221 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
    14281 cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
    14341 tatctgctcc ctgcttgtgt gttggaggtc gctga
//
