LOCUS       sequence-383147-       11629 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Piggybac Tet-ON plasmid for inducing hiPSCs into photoreceptor-like 
            cells via CRX and NeuroD1 expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11629)
  AUTHORS   Otsuka Y, Imamura K, Oishi A, Kondo T, Suga M, Yada Y, Shibukawa R, 
            Okanishi Y, Sagara Y, Tsukita K, Tsujikawa A, Inoue H
  TITLE     One-step induction of photoreceptor-like cells from human iPSCs by 
            delivering transcription factors.
  JOURNAL   iScience. 2022 Mar 8;25(4):103987. doi: 10.1016/j.isci.2022.103987. 
            eCollection 2022 Apr 15.
  PUBMED    35330684
REFERENCE   2  (bases 1 to 11629)
  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: "iScience.";
            date: "2022-03-8"; volume: "25(4):103987. doi"; pages: " 
            10.1016/j.isci.2022.103987. eCollection 2022 Apr 15"
COMMENT     Sequence Label: PB-CRX-T2A-NeuroD1
FEATURES             Location/Qualifiers
     source          1..11629
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(28..456)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(138..159)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     350..369
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     583..605
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     597..614
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     598..614
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        621..639
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   706..768
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    1140..1158
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1177..1195
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1214..1232
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1251..1269
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1288..1306
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1325..1343
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1362..1380
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1399..1417
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1439..1463
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    1542..1566
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2501..2554
                     /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"
     protein_bind    complement(3635..3659)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3774..3793)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3839..3894
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3893..3912)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     enhancer        4337..4716
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4831..6136
                     /label=rEF-1-alpha promoter
                     /note="strong constitutive promoter for rat elongation 
                     factor EF-1-alpha"
     misc_feature    5173..6104
                     /label=rEF-1-alpha intron
                     /note="intron upstream of the start codon of rat 
                     EF-1-alpha"
     CDS             join(6153..6770,6771..6776,6777..6812,6813..6815,6816..6851,
                     6852..6854,6855..6890,6891..6899)
                     /codon_start=1
                     /product="improved tetracycline-controlled transactivator"
                     /label=rtTA-Advanced
                     /note="In the Tet-On(R) system, rtTA-Advanced binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the presence of tetracycline or doxycycline."
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     misc_feature    6926..7370
                     /label=FMDV IRES
                     /note="internal ribosome entry site (IRES) of the 
                     foot-and-mouth disease virus"
     promoter        7403..7449
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        7441..7459
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7469..8263
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             7469..8263
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7523..7542)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8133..8152
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    8310..8882
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
     repeat_region   9256..9290
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(9439..9457)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(9478..9494)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9478..9494)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(9491..9513)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    9502..9518
                     /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(9526..9532,9533..9550,9551..9556))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9571..9592
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9709..9726)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9880..10468)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9960..9979)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10639..11430,11431..11499))
                     /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     11262..11281
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11500..11604)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
       61 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
      121 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
      181 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
      241 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
      301 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
      361 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
      421 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
      481 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
      541 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
      601 aaacgacggc cagtgaattg taatacgact cactataggg cgaattggag ctcggtattc
      661 acgacagcag gctgaataat aaaaaaatta gaaactatta tttaacccta gaaagataat
      721 catattgtga cgtacgttaa agataatcat gcgtaaaatt gacgcatgtg ttttatcggt
      781 ctgtatatcg aggtttattt attaatttga atagatatta agttttatta tatttacact
      841 tacatactaa taataaattc aacaaacaat ttatttatgt ttatttattt attaaaaaaa
      901 aacaaaaact caaaatttct tctataaagt aacaaaactt ttaaacattc tctcttttac
      961 aaaaataaac ttattttgta ctttaaaaac agtcatgttg tattataaaa taagtaatta
     1021 gcttaaccta tacataatag aaacaaatta tacttattag tcagtcagaa acaactttgg
     1081 cacatatcaa tattatgctc tcgttaatta actcgaggga tccaagctgg ccgcacgtct
     1141 ccctatcagt gatagagaag tcgacacgtc tcgagctccc tatcagtgat agagaaggta
     1201 cgtctagaac gtctccctat cagtgataga gaagtcgaca cgtctcgagc tccctatcag
     1261 tgatagagaa ggtacgtcta gaacgtctcc ctatcagtga tagagaagtc gacacgtctc
     1321 gagctcccta tcagtgatag agaaggtacg tctagaacgt ctccctatca gtgatagaga
     1381 agtcgacacg tctcgagctc cctatcagtg atagagaagg taccccctat ataagcagag
     1441 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
     1501 gaagacaccg ggaccgatcc agcctccgcg gtggcggcat cacaagtttg tacaaaaaag
     1561 caggctttaa aggaaccaat tcagtcgact ggatccggta ccgatgatgg cgtatatgaa
     1621 cccggggccc cactattctg tcaacgcctt ggccctaagt ggccccagtg tggatctgat
     1681 gcaccaggct gtgccctacc caagcgcccc caggaagcag cggcgggagc gcaccacctt
     1741 cacccggagc caactggagg agctggaggc actgtttgcc aagacccagt acccagacgt
     1801 ctatgcccgt gaggaggtgg ctctgaagat caatctgcct gagtccaggg ttcaggtttg
     1861 gttcaagaac cggagggcta aatgcaggca gcagcgacag cagcagaaac agcagcagca
     1921 gcccccaggg ggccaggcca aggcccggcc tgccaagagg aaggcgggca cgtccccaag
     1981 accctccaca gatgtgtgtc cagaccctct gggcatctca gattcctaca gtccccctct
     2041 gcccggcccc tcaggctccc caaccacggc agtggccact gtgtccatct ggagcccagc
     2101 ctcagagtcc cctttgcctg aggcgcagcg ggctgggctg gtggcctcag ggccgtctct
     2161 gacctccgcc ccctatgcca tgacctacgc cccggcctcc gctttctgct cttccccctc
     2221 cgcctatggg tctccgagct cctatttcag cggcctagac ccctaccttt ctcccatggt
     2281 gccccagcta gggggcccgg ctcttagccc cctctctggc ccctccgtgg gaccttccct
     2341 ggcccagtcc cccacctccc tatcaggcca gagctatggc gcctacagcc ccgtggatag
     2401 cttggaattc aaggacccca cgggcacctg gaaattcacc tacaatccca tggaccctct
     2461 ggactacaag gatcagagtg cctggaagtt tcagatcttg gagggcagag gcagcctgct
     2521 gacctgcggc gacgtggagg aaaaccctgg ccctatgacc aaatcgtaca gcgagagtgg
     2581 gctgatgggc gagcctcagc cccaaggtcc tccaagctgg acagacgagt gtctcagttc
     2641 tcaggacgag gagcacgagg cagacaagaa ggaggacgac ctcgaaacca tgaacgcaga
     2701 ggaggactca ctgaggaacg ggggagagga ggaggacgaa gatgaggacc tggaagagga
     2761 ggaagaagag gaagaggagg atgacgatca aaagcccaag agacgcggcc ccaaaaagaa
     2821 gaagatgact aaggctcgcc tggagcgttt taaattgaga cgcatgaagg ctaacgcccg
     2881 ggagcggaac cgcatgcacg gactgaacgc ggcgctagac aacctgcgca aggtggtgcc
     2941 ttgctattct aagacgcaga agctgtccaa aatcgagact ctgcgcttgg ccaagaacta
     3001 catctgggct ctgtcggaga tcctgcgctc aggcaaaagc ccagacctgg tctccttcgt
     3061 tcagacgctt tgcaagggct tatcccaacc caccaccaac ctggttgcgg gctgcctgca
     3121 actcaatcct cggacttttc tgcctgagca gaaccaggac atgccccccc acctgccgac
     3181 ggccagcgct tccttccctg tacaccccta ctcctaccag tcgcctgggc tgcccagtcc
     3241 gccttacggt accatggaca gctcccatgt cttccacgtt aagcctccgc cgcacgccta
     3301 cagcgcagcg ctggagccct tctttgaaag ccctctgact gattgcacca gcccttcctt
     3361 tgatggaccc ctcagcccgc cgctcagcat caatggcaac ttctctttca aacacgaacc
     3421 gtccgccgag tttgagaaaa attatgcctt taccatgcac tatcctgcag cgacactggc
     3481 aggggcccaa agccacggat caatcttctc aggcaccgct gcccctcgct gcgagatccc
     3541 catagacaat attatgtcct tcgatagcca ttcacatcat gagcgagtca tgagtgccca
     3601 gctcaatgcc atatttcatg attaggatat ctagacccag ctttcttgta caaagtggtg
     3661 atggccgctc tagcgaaggg atcccccatc gaattcactc ctcaggtgca ggctgcctat
     3721 cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt
     3781 ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa
     3841 taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga
     3901 aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg
     3961 gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag
     4021 tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt
     4081 tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc
     4141 ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc
     4201 cctcttctct tatggagatc cctcgacctg caggaattcg attacttaag cctaggatcc
     4261 agatctgtac attcgaatcg atgaattcgt taaccatgga agcttctgca gcggccgcac
     4321 tagcggtggc ggcctcgaca ttgattattg actagttatt aatagtaatc aattacgggg
     4381 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     4441 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     4501 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     4561 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     4621 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     4681 cagtacatct acgtattagt catcgctatt accatgatga tgcggttttg gcagtacatc
     4741 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     4801 aatgggagtt tgttttgact agtggagccg agagtaattc atacaaaagg agggatcgcc
     4861 ttcgcaaggg gagagcccag ggaccgtccc taaattctca cagacccaaa tccctgtagc
     4921 cgccccacga cagcgcgagg agcatgcgcc cagggctgag cgcgggtaga tcagagcaca
     4981 caagctcaca gtccccggcg gtggggggag gggcgcgctg agcgggggcc agggagctgg
     5041 cgcggggcaa actgggaaag tggtgtcgtg tgctggctcc gccctcttcc cgagggtggg
     5101 ggagaacggt atataagtgc ggtagtcgcc ttggacgttc tttttcgcaa cgggtttgcc
     5161 gtcagaacgc aggtgagtgg cgggtgtggc ttccgcgggc cccggagctg gagccctgct
     5221 ctgagcgggc cgggctgata tgcgagtgtc gtccgcaggg tttagctgtg agcattccca
     5281 cttcgagtgg cgggcggtgc gggggtgaga gtgcgaggcc tagcggcaac cccgtagcct
     5341 cgcctcgtgt ccggcttgag gcctagcgtg gtgtccgccg ccgcgtgcca ctccggccgc
     5401 actatgcgtt ttttgtcctt gctgccctcg attgccttcc agcagcatgg gctaacaaag
     5461 ggagggtgtg gggctcactc ttaaggagcc catgaagctt acgttggata ggaatggaag
     5521 ggcaggaggg gcgactgggg cccgcccgcc ttcggagcac atgtccgacg ccacctggat
     5581 ggggcgaggc ctgtggcttt ccgaagcaat cgggcgtgag tttagcctac ctgggccatg
     5641 tggccctagc actgggcacg gtctggcctg gcggtgccgc gttcccttgc ctcccaacaa
     5701 gggtgaggcc gtcccgcccg gcaccagttg cttgcgcgga aagatggccg ctcccggggc
     5761 cctgttgcaa ggagctcaaa atggaggacg cggcagcccg gtggagcggg cgggtgagtc
     5821 acccacacaa aggaagaggg ccttgcccct cgccggccgc tgcttcctgt gaccccgtgg
     5881 tctatcggcc gcatagtcac ctcgggcttc tcttgagcac cgctcgtcgc ggcgggggga
     5941 ggggatctaa tggcgttgga gtttgttcac atttggtggg tggagactag tcaggccagc
     6001 ctggcgctgg aagtcattct tggaatttgc ccctttgagt ttggagcgag gctaattctc
     6061 aagcctctta gcggttcaaa ggtattttct aaacccgttt ccaggtgttg tgaaagccac
     6121 cgctaattca aagcaatccg gagtaattca ccatgtctag actggacaag agcaaagtca
     6181 taaacggcgc tctggaatta ctcaatggag tcggtatcga aggcctgacg acaaggaaac
     6241 tcgctcaaaa gctgggagtt gagcagccta ccctgtactg gcacgtgaag aacaagcggg
     6301 ccctgctcga tgccctgcca atcgagatgc tggacaggca tcatacccac ttctgccccc
     6361 tggaaggcga gtcatggcaa gactttctgc ggaacaacgc caagtcattc cgctgtgctc
     6421 tcctctcaca tcgcgacggg gctaaagtgc atctcggcac ccgcccaaca gagaaacagt
     6481 acgaaaccct ggaaaatcag ctcgcgttcc tgtgtcagca aggcttctcc ctggagaacg
     6541 cactgtacgc tctgtccgcc gtgggccact ttacactggg ctgcgtattg gaggaacagg
     6601 agcatcaagt agcaaaagag gaaagagaga cacctaccac cgattctatg cccccacttc
     6661 tgagacaagc aattgagctg ttcgaccggc agggagccga acctgccttc cttttcggcc
     6721 tggaactaat catatgtggc ctggagaaac agctaaagtg cgaaagcggc gggccggccg
     6781 acgcccttga cgattttgac ttagacatgc tcccagccga tgcccttgac gactttgacc
     6841 ttgatatgct gcctgctgac gctcttgacg attttgacct tgacatgctc cccgggtaac
     6901 taagtaagga tcccgtacgc ctaggagcag gtttccccaa tgacacaaaa cgtgcaactt
     6961 gaaactccgc ctggtctttc caggtctaga ggggtaacac tttgtactgc gtttggctcc
     7021 acgctcgatc cactggcgag tgttagtaac agcactgttg cttcgtagcg gagcatgacg
     7081 gccgtgggaa ctcctccttg gtaacaagga cccacggggc caaaagccac gcccacacgg
     7141 gcccgtcatg tgtgcaaccc cagcacggcg actttactgc gaaacccact ttaaagtgac
     7201 attgaaactg gtacccacac actggtgaca ggctaaggat gcccttcagg taccccgagg
     7261 taacacgcga cactcgggat ctgagaaggg gactggggct tctataaaag cgctcggttt
     7321 aaaaagcttc tatgcctgaa taggtgaccg gaggtcggca cctttccttt gcaattactg
     7381 accctatgaa tacactgact gtttgacaat taatcatcgg catagtatat cggcatagta
     7441 taatacgact cactatagga gggccaccat gattgaacaa gatggattgc acgcaggttc
     7501 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     7561 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     7621 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     7681 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     7741 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     7801 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     7861 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     7921 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     7981 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgacac atggcgatgc
     8041 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     8101 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     8161 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     8221 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     8281 gaaatgaccg accaagcgaa ttcgctagga ttatccctaa tacctgccac cccactctta
     8341 atcagtggtg gaagaacggt ctcagaactg tttgtttcaa ttggccattt aagtttagta
     8401 gtaaaagact ggttaatgat aacaatgcat cgtaaaacct tcagaaggaa aggagaatgt
     8461 tttgtggacc actttggttt tcttttttgc gtgtggcagt tttaagttat tagtttttaa
     8521 aatcagtact ttttaatgga aacaacttga ccaaaaattt gtcacagaat tttgagaccc
     8581 attaaaaaag ttaaatgaga aacctgtgtg ttcctttggt caacaccgag acatttaggt
     8641 gaaagacatc taattctggt tttacgaatc tggaaacttc ttgaaaatgt aattcttgag
     8701 ttaacacttc tgggtggaga atagggttgt tttcccccca cataattgga aggggaagga
     8761 atatcattta aagctatggg agggttgctt tgattacaac actggagaga aatgcagcat
     8821 gttgctgatt gcctgtcact aaaacaggcc aaaaactgag tccttgggtt gcatagaaag
     8881 ctgccaaacg ctagccaaca agctcgtcat cgctttgcag aagagcagag aggatatgct
     8941 catcgtctaa agaactaccc attttattat atattagtca cgatatctat aacaagaaaa
     9001 tatatatata ataagttatc acgtaagtag aacatgaaat aacaatataa ttatcgtatg
     9061 agttaaatct taaaagtcac gtaaaagata atcatgcgtc attttgactc acgcggtcgt
     9121 tatagttcaa aatcagtgac acttaccgca ttgacaagca cgcctcacgg gagctccaag
     9181 cggcgactga gatgtcctaa atgcacagcg acggattcgc gctatttaga aagagagagc
     9241 aatatttcaa gaatgcatgc gtcaatttta cgcagactat ctttctaggg ttaaaaaaga
     9301 tttgcgcttt actcgaccta aactttaaac acgtcataga atcttcgttt gacaaaaacc
     9361 acattgtggc caagctgtgt gacgcgacgc gcgctaaaga atggcaaacc aagtcgcgcg
     9421 aggtaccagc ttttgttccc tttagtgagg gttaatttcg agcttggcgt aatcatggtc
     9481 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     9541 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     9601 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     9661 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     9721 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     9781 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     9841 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     9901 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     9961 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
    10021 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
    10081 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
    10141 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
    10201 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
    10261 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
    10321 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
    10381 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
    10441 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
    10501 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
    10561 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
    10621 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
    10681 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
    10741 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
    10801 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
    10861 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
    10921 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
    10981 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
    11041 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
    11101 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
    11161 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
    11221 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
    11281 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
    11341 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
    11401 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
    11461 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
    11521 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
    11581 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacct
//
