LOCUS       sequence-356777-       10192 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TetO-Ascl1 in a PiggyBac vector with neomycin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10192)
  AUTHORS   Ng YH, Chanda S, Janas JA, Yang N, Kokubu Y, Sudhof TC, Wernig M
  TITLE     Efficient generation of dopaminergic induced neuronal cells with 
            midbrain characteristics.
  JOURNAL   Stem Cell Reports. 2021 Jul 13;16(7):1763-1776. doi: 
            10.1016/j.stemcr.2021.05.017. Epub 2021 Jun 24.
  PUBMED    34171286
REFERENCE   2  (bases 1 to 10192)
  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.stemcr.2021.05.017"; journalName: "Stem Cell Reports"; 
            date: "2021-07-13- 13"; volume: "16"; issue: "7"; pages: "1763-1776"
COMMENT     Sequence Label: PiggyBac-TetO-Ascl1-neomycin
FEATURES             Location/Qualifiers
     source          1..10192
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     41..63
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     55..72
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     56..72
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        79..97
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   164..226
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     protein_bind    598..616
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    635..653
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    672..690
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    709..727
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    746..764
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    783..801
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    857..875
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     897..921
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    1001..1025
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1044..1053
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1828..1883
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1882..1901)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     enhancer        2358..2737
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2852..4157
                     /label=rEF-1-alpha promoter
                     /note="strong constitutive promoter for rat elongation 
                     factor EF-1-alpha"
     misc_feature    3194..4125
                     /label=rEF-1-alpha intron
                     /note="intron upstream of the start codon of rat 
                     EF-1-alpha"
     CDS             join(4174..4791,4792..4797,4798..4833,4834..4836,4837..4872,
                     4873..4875,4876..4911,4912..4920)
                     /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    4947..5391
                     /label=FMDV IRES
                     /note="internal ribosome entry site (IRES) of the 
                     foot-and-mouth disease virus"
     promoter        5424..5470
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        5462..5480
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5490..6284
                     /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             5490..6284
                     /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(5544..5563)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6154..6173
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6331..6903
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
     repeat_region   7277..7311
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     promoter        complement(7460..7478)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7499..7515)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7499..7515)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7512..7534)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7523..7539
                     /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(7547..7553,7554..7571,7572..7577))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7592..7613
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7730..7747)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7901..8489)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7981..8000)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8660..9451,9452..9520))
                     /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     9283..9302
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9521..9625)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(9651..10106)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9788..9809)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     10000..10019
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
       61 acgacggcca gtgaattgta atacgactca ctatagggcg aattggagct cggtattcac
      121 gacagcaggc tgaataataa aaaaattaga aactattatt taaccctaga aagataatca
      181 tattgtgacg tacgttaaag ataatcatgc gtaaaattga cgcatgtgtt ttatcggtct
      241 gtatatcgag gtttatttat taatttgaat agatattaag ttttattata tttacactta
      301 catactaata ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaaa
      361 caaaaactca aaatttcttc tataaagtaa caaaactttt aaacattctc tcttttacaa
      421 aaataaactt attttgtact ttaaaaacag tcatgttgta ttataaaata agtaattagc
      481 ttaacctata cataatagaa acaaattata cttattagtc agtcagaaac aactttggca
      541 catatcaata ttatgctctc gttaattaac tcgagggatc caagctggcc gcacgtctcc
      601 ctatcagtga tagagaagtc gacacgtctc gagctcccta tcagtgatag agaaggtacg
      661 tctagaacgt ctccctatca gtgatagaga agtcgacacg tctcgagctc cctatcagtg
      721 atagagaagg tacgtctaga acgtctccct atcagtgata gagaagtcga cacgtctcga
      781 gctccctatc agtgatagag aaggtacgtc tagaacgtct ccctatcagt gatagagaag
      841 tcgacacgtc tcgagctccc tatcagtgat agagaaggta ccccctatat aagcagagct
      901 cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga cctccataga
      961 agacaccggg accgatccag cctccgcggt ggcggccatc acaagtttgt acaaaaaagc
     1021 aggctccgcg gccgccgacc ggggccacca tggagagctc tggcaagatg gagagtggag
     1081 ccggccagca gccgcagccc ccgcagccct tcctgcctcc cgcagcctgc ttctttgcga
     1141 ccgcggcggc ggcggcagcg gcggcggccg cggcagctca gagcgcgcag cagcaacagc
     1201 cgcaggcgcc gccgcagcag gcgccgcagc tgagcccggt ggccgacagc cagccctcag
     1261 ggggcggtca caagtcagcg gccaagcagg tcaagcgcca gcgctcgtcc tctccggaac
     1321 tgatgcgctg caaacgccgg ctcaacttca gcggcttcgg ctacagcctg ccacagcagc
     1381 agccggccgc cgtggcgcgc cgcaacgagc gcgagcgcaa ccgggtcaag ttggtcaacc
     1441 tgggctttgc caccctccgg gagcatgtcc ccaacggcgc ggccaacaag aagatgagca
     1501 aggtggagac gctgcgctcg gcggtcgagt acatccgcgc gctgcagcag ctgctggacg
     1561 agcacgacgc ggtgagcgct gcctttcagg cgggcgtcct gtcgcccacc atctccccca
     1621 actactccaa cgacttgaac tctatggcgg gttctccggt ctcgtcctac tcctccgacg
     1681 agggatccta cgaccctctt agcccagagg aacaagagct gctggacttt accaactggt
     1741 tctaaaccgg tcgggcgcgc cgatcttttc cctctgccaa aaattatggg gacatcatga
     1801 agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt
     1861 gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac
     1921 atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga
     1981 acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct
     2041 tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat
     2101 ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc
     2161 tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgacctgc
     2221 aggaattcga ttacttaagc ctaggatcca gatctgtaca ttcgaatcga tgaattcgtt
     2281 aaccatggaa gcttctgcaa agggcgaatt ctgcagatat ccatcacact ggcggccgca
     2341 ctagcggtgg cggcctcgac attgattatt gactagttat taatagtaat caattacggg
     2401 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     2461 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     2521 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     2581 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     2641 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     2701 gcagtacatc tacgtattag tcatcgctat taccatgatg atgcggtttt ggcagtacat
     2761 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     2821 caatgggagt ttgttttgac tagtggagcc gagagtaatt catacaaaag gagggatcgc
     2881 cttcgcaagg ggagagccca gggaccgtcc ctaaattctc acagacccaa atccctgtag
     2941 ccgccccacg acagcgcgag gagcatgcgc ccagggctga gcgcgggtag atcagagcac
     3001 acaagctcac agtccccggc ggtgggggga ggggcgcgct gagcgggggc cagggagctg
     3061 gcgcggggca aactgggaaa gtggtgtcgt gtgctggctc cgccctcttc ccgagggtgg
     3121 gggagaacgg tatataagtg cggtagtcgc cttggacgtt ctttttcgca acgggtttgc
     3181 cgtcagaacg caggtgagtg gcgggtgtgg cttccgcggg ccccggagct ggagccctgc
     3241 tctgagcggg ccgggctgat atgcgagtgt cgtccgcagg gtttagctgt gagcattccc
     3301 acttcgagtg gcgggcggtg cgggggtgag agtgcgaggc ctagcggcaa ccccgtagcc
     3361 tcgcctcgtg tccggcttga ggcctagcgt ggtgtccgcc gccgcgtgcc actccggccg
     3421 cactatgcgt tttttgtcct tgctgccctc gattgccttc cagcagcatg ggctaacaaa
     3481 gggagggtgt ggggctcact cttaaggagc ccatgaagct tacgttggat aggaatggaa
     3541 gggcaggagg ggcgactggg gcccgcccgc cttcggagca catgtccgac gccacctgga
     3601 tggggcgagg cctgtggctt tccgaagcaa tcgggcgtga gtttagccta cctgggccat
     3661 gtggccctag cactgggcac ggtctggcct ggcggtgccg cgttcccttg cctcccaaca
     3721 agggtgaggc cgtcccgccc ggcaccagtt gcttgcgcgg aaagatggcc gctcccgggg
     3781 ccctgttgca aggagctcaa aatggaggac gcggcagccc ggtggagcgg gcgggtgagt
     3841 cacccacaca aaggaagagg gccttgcccc tcgccggccg ctgcttcctg tgaccccgtg
     3901 gtctatcggc cgcatagtca cctcgggctt ctcttgagca ccgctcgtcg cggcgggggg
     3961 aggggatcta atggcgttgg agtttgttca catttggtgg gtggagacta gtcaggccag
     4021 cctggcgctg gaagtcattc ttggaatttg cccctttgag tttggagcga ggctaattct
     4081 caagcctctt agcggttcaa aggtattttc taaacccgtt tccaggtgtt gtgaaagcca
     4141 ccgctaattc aaagcaatcc ggagtaattc accatgtcta gactggacaa gagcaaagtc
     4201 ataaacggcg ctctggaatt actcaatgga gtcggtatcg aaggcctgac gacaaggaaa
     4261 ctcgctcaaa agctgggagt tgagcagcct accctgtact ggcacgtgaa gaacaagcgg
     4321 gccctgctcg atgccctgcc aatcgagatg ctggacaggc atcataccca cttctgcccc
     4381 ctggaaggcg agtcatggca agactttctg cggaacaacg ccaagtcatt ccgctgtgct
     4441 ctcctctcac atcgcgacgg ggctaaagtg catctcggca cccgcccaac agagaaacag
     4501 tacgaaaccc tggaaaatca gctcgcgttc ctgtgtcagc aaggcttctc cctggagaac
     4561 gcactgtacg ctctgtccgc cgtgggccac tttacactgg gctgcgtatt ggaggaacag
     4621 gagcatcaag tagcaaaaga ggaaagagag acacctacca ccgattctat gcccccactt
     4681 ctgagacaag caattgagct gttcgaccgg cagggagccg aacctgcctt ccttttcggc
     4741 ctggaactaa tcatatgtgg cctggagaaa cagctaaagt gcgaaagcgg cgggccggcc
     4801 gacgcccttg acgattttga cttagacatg ctcccagccg atgcccttga cgactttgac
     4861 cttgatatgc tgcctgctga cgctcttgac gattttgacc ttgacatgct ccccgggtaa
     4921 ctaagtaagg atcccgtacg cctaggagca ggtttcccca atgacacaaa acgtgcaact
     4981 tgaaactccg cctggtcttt ccaggtctag aggggtaaca ctttgtactg cgtttggctc
     5041 cacgctcgat ccactggcga gtgttagtaa cagcactgtt gcttcgtagc ggagcatgac
     5101 ggccgtggga actcctcctt ggtaacaagg acccacgggg ccaaaagcca cgcccacacg
     5161 ggcccgtcat gtgtgcaacc ccagcacggc gactttactg cgaaacccac tttaaagtga
     5221 cattgaaact ggtacccaca cactggtgac aggctaagga tgcccttcag gtaccccgag
     5281 gtaacacgcg acactcggga tctgagaagg ggactggggc ttctataaaa gcgctcggtt
     5341 taaaaagctt ctatgcctga ataggtgacc ggaggtcggc acctttcctt tgcaattact
     5401 gaccctatga atacactgac tgtttgacaa ttaatcatcg gcatagtata tcggcatagt
     5461 ataatacgac tcactatagg agggccacca tgattgaaca agatggattg cacgcaggtt
     5521 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     5581 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     5641 ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta tcgtggctgg
     5701 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     5761 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     5821 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     5881 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5941 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     6001 tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgaca catggcgatg
     6061 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     6121 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     6181 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     6241 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     6301 cgaaatgacc gaccaagcga attcgctagg attatcccta atacctgcca ccccactctt
     6361 aatcagtggt ggaagaacgg tctcagaact gtttgtttca attggccatt taagtttagt
     6421 agtaaaagac tggttaatga taacaatgca tcgtaaaacc ttcagaagga aaggagaatg
     6481 ttttgtggac cactttggtt ttcttttttg cgtgtggcag ttttaagtta ttagttttta
     6541 aaatcagtac tttttaatgg aaacaacttg accaaaaatt tgtcacagaa ttttgagacc
     6601 cattaaaaaa gttaaatgag aaacctgtgt gttcctttgg tcaacaccga gacatttagg
     6661 tgaaagacat ctaattctgg ttttacgaat ctggaaactt cttgaaaatg taattcttga
     6721 gttaacactt ctgggtggag aatagggttg ttttcccccc acataattgg aaggggaagg
     6781 aatatcattt aaagctatgg gagggttgct ttgattacaa cactggagag aaatgcagca
     6841 tgttgctgat tgcctgtcac taaaacaggc caaaaactga gtccttgggt tgcatagaaa
     6901 gctgccaaac gctagccaac aagctcgtca tcgctttgca gaagagcaga gaggatatgc
     6961 tcatcgtcta aagaactacc cattttatta tatattagtc acgatatcta taacaagaaa
     7021 atatatatat aataagttat cacgtaagta gaacatgaaa taacaatata attatcgtat
     7081 gagttaaatc ttaaaagtca cgtaaaagat aatcatgcgt cattttgact cacgcggtcg
     7141 ttatagttca aaatcagtga cacttaccgc attgacaagc acgcctcacg ggagctccaa
     7201 gcggcgactg agatgtccta aatgcacagc gacggattcg cgctatttag aaagagagag
     7261 caatatttca agaatgcatg cgtcaatttt acgcagacta tctttctagg gttaaaaaag
     7321 atttgcgctt tactcgacct aaactttaaa cacgtcatag aatcttcgtt tgacaaaaac
     7381 cacattgtgg ccaagctgtg tgacgcgacg cgcgctaaag aatggcaaac caagtcgcgc
     7441 gaggtaccag cttttgttcc ctttagtgag ggttaatttc gagcttggcg taatcatggt
     7501 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     7561 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     7621 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     7681 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     7741 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     7801 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     7861 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     7921 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     7981 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     8041 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     8101 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     8161 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     8221 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     8281 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     8341 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     8401 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     8461 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     8521 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     8581 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     8641 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     8701 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     8761 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     8821 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     8881 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     8941 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     9001 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     9061 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     9121 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     9181 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     9241 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     9301 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     9361 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     9421 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     9481 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     9541 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     9601 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag
     9661 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     9721 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     9781 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     9841 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     9901 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     9961 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
    10021 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
    10081 gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca actgttggga
    10141 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg ga
//
