LOCUS       sequence-362358-       12343 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Donor plasmid for stable integration of firefly luciferase NMD(+) 
            PTC39 reporter at AAVS1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12343)
  AUTHORS   Udy DB, Bradley RK
  TITLE     Nonsense-mediated mRNA decay uses complementary mechanisms to 
            suppress mRNA and protein accumulation.
  JOURNAL   Life Sci Alliance. 2021 Dec 8;5(3). pii: 5/3/e202101217. doi: 
            10.26508/lsa.202101217. Print 2022 Mar.
  PUBMED    34880103
REFERENCE   2  (bases 1 to 12343)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Life Sci 
            Alliance."; date: "2021-12-8"; volume: "5(3). pii: 5/3/e202101217. 
            doi"; pages: " 10.26508/lsa.202101217. Print 2022 Mar"
COMMENT     Sequence Label: AAVS1-TetOn-3XFLAG-firefly-beta-globin-PTC39-A...
FEATURES             Location/Qualifiers
     source          1..12343
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        242..260
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    295..1098
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     polyA_signal    1129..1210
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(1327..2073)
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     promoter        complement(2092..2602)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     polyA_signal    2622..2670
                     /note="synthetic polyadenylation signal"
     misc_feature    2684..2775
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     2724..2743
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     promoter        2779..3146
                     /label=TRE3GS promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription 
                     factors"
     protein_bind    2787..2805
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2823..2841
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2859..2877
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2895..2913
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2931..2949
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2967..2985
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3003..3021
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             3181..3246
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3265..4911
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="EDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDAH
                     IEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAPA
                     NDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQSM
                     YTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHAR
                     DPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYKI
                     QSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGYG
                     LTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMSG
                     YVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESILL
                     QHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVVF
                     VDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(3328..3346)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     protein_bind    4939..4963
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    6366..6447
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        6469..6979
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             7001..7600
                     /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(7001..7020)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7497..7517
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7607..7831
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    7845..8681
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     primer_bind     complement(8715..8734)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(8716..8734)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8741..8758)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(8741..8757)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8750..8772)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             8895..9197
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     9076..9095
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             9546..10340
                     /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             9546..10340
                     /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(9600..9619)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     10210..10229
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(join(10590..11381,11382..11450))
                     /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     11213..11232
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      11574..12162
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     12063..12082
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctca
      241 gaattaaccc tcactaaagg gactagtcct gcaggtttaa acgaattcgc cctttgcttt
      301 ctctgaccag cattctctcc cctgggcctg tgccgctttc tgtctgcagc ttgtggcctg
      361 ggtcacctct acggctggcc cagatccttc cctgccgcct ccttcaggtt ccgtcttcct
      421 ccactccctc ttccccttgc tctctgctgt gttgctgccc aaggatgctc tttccggagc
      481 acttccttct cggcgctgca ccacgtgatg tcctctgagc ggatcctccc cgtgtctggg
      541 tcctctccgg gcatctctcc tccctcaccc aaccccatgc cgtcttcact cgctgggttc
      601 ccttttcctt ctccttctgg ggcctgtgcc atctctcgtt tcttaggatg gccttctccg
      661 acggatgtct cccttgcgtc ccgcctcccc ttcttgtagg cctgcatcat caccgttttt
      721 ctggacaacc ccaaagtacc ccgtctccct ggctttagcc acctctccat cctcttgctt
      781 tctttgcctg gacaccccgt tctcctgtgg attcgggtca cctctcactc ctttcatttg
      841 ggcagctccc ctacccccct tacctctcta gtctgtgcta gctcttccag ccccctgtca
      901 tggcatcttc caggggtccg agagctcagc tagtcttctt cctccaaccc gggcccctat
      961 gtccacttca ggacagcatg tttgctgcct ccagggatcc tgtgtccccg agctgggacc
     1021 accttatatt cccagggccg gttaatgtgg ctctggttct gggtactttt atctgtcccc
     1081 tccaccccac agtggggcaa gctgagtcag tgagcgagga agctcggggc agtgaaaaaa
     1141 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     1201 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     1261 ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatcctct
     1321 ggagatttac ccggggagca tgtcaaggtc aaaatcgtca agagcgtcag caggcagcat
     1381 atcaaggtca aagtcgtcaa gggcatcggc tgggagcatg tctaagtcaa aatcgtcaag
     1441 ggcgtcggtc ggcccgccgc tttcgcactt tagctgtttc tccaggccac atatgattag
     1501 ttccaggccg aaaaggaagg caggttcggc tccctgccgg tcgaacagct caattgcttg
     1561 tttcagaagt gggggcatag aatcggtggt aggtgtctct ctttcctctt ttgctacttg
     1621 atgctcctgt tcctccaata cgcagcccag tgtaaagtgg cccacggcgg acagagcgta
     1681 cagtgcgttc tccagggaga agccttgctg acacaggaac gcgagctgat tttccagggt
     1741 ttcgtactgt ttctctgttg ggcgggtgcc gagatgcact ttagccccgt cgcgatgtga
     1801 gaggagagca cagcggtatg acttggcgtt gttccgcaga aagtcttgcc atgactcgcc
     1861 ttccaggggg caggagtggg tatgatgcct gtccagcatc tcgattggca gggcatcgag
     1921 cagggcccgc ttgttcttca cgtgccagta cagggtaggc tgctcaactc ccagcttttg
     1981 agcgagtttc cttgtcgtca ggccttcgat accgactcca ttgagtaatt ccagagcaga
     2041 gtttatgact ttgctcttgt ccagtctaga catggtaatt cgatgatccc cctggggaga
     2101 gaggtcggtg attcggtcaa cgagggagcc gactgccgac gtgcgctccg gaggcttgca
     2161 gaatgcggaa caccgcgcgg gcaggaacag ggcccacact accgccccac accccgcctc
     2221 ccgcaccgcc ccttcccggc cgctgctctc ggcgcgccct gctgagcagc cgctattggc
     2281 cacagcccat cgcggtcggc gcgctgccat tgctccctgg cgctgtccgt ctgcgagggt
     2341 actagtgaga cgtgcggctt ccgtttgtca cgtccggcac gccgcgaacc gcaaggaacc
     2401 ttcccgactt aggggcggag caggaagcgt cgccgggggg cccacaaggg tagcggcgaa
     2461 gatccgggtg acgctgcgaa cggacgtgaa gaatgtgcga gacccagggt cggcgccgct
     2521 gcgtttcccg gaaccacgcc cagagcagcc gcgtccctgc gcaaacccag ggctgccttg
     2581 gaaaaggcgc aaccccaacc ccgtggaatt atcacctcga gaataaaata tctttatttt
     2641 cattacatct gtgtgttggt tttttgtgtg aatcgatagt actaacatac gctctccatc
     2701 aaaacaaaac gaaacaaaac aaactagcaa aataggctgt ccccagtgca agtgcaggtg
     2761 ccagaacatt tctctctcga gtttactccc tatcagtgat agagaacgta tgaagagttt
     2821 actccctatc agtgatagag aacgtatgca gactttactc cctatcagtg atagagaacg
     2881 tataaggagt ttactcccta tcagtgatag agaacgtatg accagtttac tccctatcag
     2941 tgatagagaa cgtatctaca gtttactccc tatcagtgat agagaacgta tatccagttt
     3001 actccctatc agtgatagag aacgtataag ctttgcttat gtaaaccagg gcgcctataa
     3061 aagagtgctg attttttgag taaacttcaa ttccacaaca cttttgtctt ataccaactt
     3121 tccgtaccac ttcctaccct cgtaaagaat tcgcggccgc aaccgtcaga attaaccatg
     3181 gactacaaag accatgacgg tgattataaa gatcatgaca tcgattacaa ggatgacgat
     3241 gacaagcttg cggccgcgaa ttcagaagac gccaaaaaca taaagaaagg cccggcgcca
     3301 ttctatccgc tggaagatgg aaccgctgga gagcaactgc ataaggctat gaagagatac
     3361 gccctggttc ctggaacaat tgcttttaca gatgcacata tcgaggtgga catcacttac
     3421 gctgagtact tcgaaatgtc cgttcggttg gcagaagcta tgaaacgata tgggctgaat
     3481 acaaatcaca gaatcgtcgt atgcagtgaa aactctcttc aattctttat gccggtgttg
     3541 ggcgcgttat ttatcggagt tgcagttgcg cccgcgaacg acatttataa tgaacgtgaa
     3601 ttgctcaaca gtatgggcat ttcgcagcct accgtggtgt tcgtttccaa aaaggggttg
     3661 caaaaaattt tgaacgtgca aaaaaagctc ccaatcatcc aaaaaattat tatcatggat
     3721 tctaaaacgg attaccaggg atttcagtcg atgtacacgt tcgtcacatc tcatctacct
     3781 cccggtttta atgaatacga ttttgtgcca gagtccttcg atagggacaa gacaattgca
     3841 ctgatcatga actcctctgg atctactggt ctgcctaaag gtgtcgctct gcctcataga
     3901 actgcctgcg tgagattctc gcatgccaga gatcctattt ttggcaatca aatcattccg
     3961 gatactgcga ttttaagtgt tgttccattc catcacggtt ttggaatgtt tactacactc
     4021 ggatatttga tatgtggatt tcgagtcgtc ttaatgtata gatttgaaga agagctgttt
     4081 ctgaggagcc ttcaggatta caagattcaa agtgcgctgc tggtgccaac cctattctcc
     4141 ttcttcgcca aaagcactct gattgacaaa tacgatttat ctaatttaca cgaaattgct
     4201 tctggtggcg ctcccctctc taaggaagtc ggggaagcgg ttgccaagag gttccatctg
     4261 ccaggtatca ggcaaggata tgggctcact gagactacat cagctattct gattacaccc
     4321 gagggggatg ataaaccggg cgcggtcggt aaagttgttc cattttttga agcgaaggtt
     4381 gtggatctgg ataccgggaa aacgctgggc gttaatcaaa gaggcgaact gtgtgtgaga
     4441 ggtcctatga ttatgtccgg ttatgtaaac aatccggaag cgaccaacgc cttgattgac
     4501 aaggatggat ggctacattc tggagacata gcttactggg acgaagacga acacttcttc
     4561 atcgttgacc gcctgaagtc tctgattaag tacaaaggct atcaggtggc tcccgctgaa
     4621 ttggaatcca tcttgctcca acaccccaac atcttcgacg caggtgtcgc aggtcttccc
     4681 gacgatgacg ccggtgaact tcccgccgcc gttgttgttt tggagcacgg aaagacgatg
     4741 acggaaaaag agatcgtgga ttacgtcgcc agtcaagtaa caaccgcgaa aaagttgcgc
     4801 ggaggagttg tgtttgtgga cgaagtaccg aaaggtctta ccggaaaact cgacgcaaga
     4861 aaaatcagag agatcctcat aaaggccaag aagggcggaa agatcgccgt ggcgaattca
     4921 tcgatagatc tgatatcaac aagtttgtac aaaaaagcag gctccgcggc cgcccccttc
     4981 accgtgcatc tgactcctga ggagaagtct gccgttactg ccctgtgggg caaggtgaac
     5041 gtggatgaag ttggtggtga ggccctgggc aggttggtat caaggttaca agacaggttt
     5101 aaggagacca atagaaactg ggcatgtgga gacagagaag actcttgggt ttctgatagg
     5161 cactgactct ctctgcctat tggtctattt tcccaccctt aggctgctgg tggtctaccc
     5221 ttggacctag aggttctttg agtcctttgg ggatctgtcc actcctgatg ctgttatggg
     5281 caaccctaag gtgaaggctc atggcaagaa agtgctcggt gcctttagtg atggcctggc
     5341 tcacctggac aacctcaagg gcacctttgc cacactgagt gagctgcact gtgacaagct
     5401 gcacgtggat cctgagaact tcagggtgag tctgatgggc acctcctggg tttccttccc
     5461 ctggctattc tgctcaacct tcctatcaga aaaaaagggg aagcgattct agggagcagt
     5521 ctccatgact gtgtgtggag tgttgacaag agttcggata ttttattctc tactcagaat
     5581 tgctgctccc cctcactctg ttctgtgttg tcatttcctc tttctttggt aagcttttaa
     5641 tttccagttg cattttacta aattaattaa gctggttatt tacttcccat cctgatatca
     5701 gcttcccctc ctcctttcct cccagtcctt ctctctctcc tctctctttc tctaatcctt
     5761 tcctttccct cagttcattt cttcttcttt gatctacgtt tgtttgtctt tttaaatatt
     5821 gccttgtaac ttgctcagag gacaaggaag atatgtccct gtttcttctc atagctctca
     5881 agaatagtag cataattggc ttttatgcca gggtgacagg ggaagaatat attttacata
     5941 taaattctgt ttgacatagg attcttataa taatttgtca gtagtttaag gttgcaaaca
     6001 aatgtctttg taaataagcc tgcagtatct ggtatttttg ctctacagtt atgttgatgg
     6061 ttcttccata ttcccacagc tcctgggcaa tatgatcgtg attgtgctgg gccaccacct
     6121 gggcaaggat ttcacccccg ctgcacaggc tgccttccag aaggtggtgg ctggagtggc
     6181 cactgccctg gcccacaagt atcactaaaa gggtgggcgc gccgacccag ctttcttgtt
     6241 ctgcagcata tgtctagagg atcataatca gccataccac atttgtagag gttttacttg
     6301 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     6361 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     6421 tcacaaataa agcatttttt tcactgccga cgtcaggtgg cacttttcgg ggttggggtt
     6481 gcgccttttc caaggcagcc ctgggtttgc gcagggacgc ggctgctctg ggcgtggttc
     6541 cgggaaacgc agcggcgccg accctgggtc tcgcacattc ttcacgtccg ttcgcagcgt
     6601 cacccggatc ttcgccgcta cccttgtggg ccccccggcg acgcttcctg ctccgcccct
     6661 aagtcgggaa ggttccttgc ggttcgcggc gtgccggacg tgacaaacgg aagccgcacg
     6721 tctcactagt accctcgcag acggacagcg ccagggagca atggcagcgc gccgaccgcg
     6781 atgggctgtg gccaatagcg gctgctcagc agggcgcgcc gagagcagcg gccgggaagg
     6841 ggcggtgcgg gaggcggggt gtggggcggt agtgtgggcc ctgttcctgc ccgcgcggtg
     6901 ttccgcattc tgcaagcctc cggagcgcac gtcggcagtc ggctccctcg ttgaccgaat
     6961 caccgacctc tctccccagg gggatccacc ggagcttacc atgaccgagt acaagcccac
     7021 ggtgcgcctc gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt
     7081 cgccgactac cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac
     7141 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     7201 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     7261 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     7321 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     7381 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     7441 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     7501 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     7561 accgcgcacc tggtgcatga cccgcaagcc cggtgcctga ggtaccctgt gccttctagt
     7621 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     7681 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     7741 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     7801 aggcatgctg gggatgcggt gggctctatg gggtaccaag ctttactagg gacaggattg
     7861 gtgacagaaa agccccatcc ttaggcctcc tccttcctag tctcctgata ttgggtctaa
     7921 cccccacctc ctgttaggca gattccttat ctggtgacac acccccattt cctggagcca
     7981 tctctctcct tgccagaacc tctaaggttt gcttacgatg gagccagaga ggatcctggg
     8041 agggagagct tggcaggggg tgggagggaa gggggggatg cgtgacctgc ccggttctca
     8101 gtggccaccc tgcgctaccc tctcccagaa cctgagctgc tctgacgcgg ctgtctggtg
     8161 cgtttcactg atcctggtgc tgcagcttcc ttacacttcc caagaggaga agcagtttgg
     8221 aaaaacaaaa tcagaataag ttggtcctga gttctaactt tggctcttca cctttctagt
     8281 ccccaattta tattgttcct ccgtgcgtca gttttacctg tgagataagg ccagtagcca
     8341 gccccgtcct ggcagggctg tggtgaggag gggggtgtcc gtgtggaaaa ctccctttgt
     8401 gagaatggtg cgtcctaggt gttcaccagg tcgtggccgc ctctactccc tttctctttc
     8461 tccatccttc tttccttaaa gagtccccag tgctatctgg gacatattcc tccgcccaga
     8521 gcagggtccc gcttccctaa ggccctgctc tgggcttctg ggtttgagtc cttggcaagc
     8581 ccaggagagg cgctcaggct tccctgtccc ccttcctcgt ccaccatctc atgcccctgg
     8641 ctctcctgcc ccttccctac aggggttcct ggctctgctc taagggcgaa ttcgcggccg
     8701 ctaaattcaa ttcgccctat agtgagtcgt attacaattc actggccgtc gttttacaac
     8761 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     8821 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     8881 gcctatacgt acggcagttt aaggtttaca cctataaaag agagagccgt tatcgtctgt
     8941 ttgtggatgt acagagtgat attattgaca cgccggggcg acggatggtg atccccctgg
     9001 ccagtgcacg tctgctgtca gataaagtct cccgtgaact ttacccggtg gtgcatatcg
     9061 gggatgaaag ctggcgcatg atgaccaccg atatggccag tgtgccggtc tccgttatcg
     9121 gggaagaagt ggctgatctc agccaccgcg aaaatgacat caaaaacgcc attaacctga
     9181 tgttctgggg aatataaatg tcaggcatga gattatcaaa aaggatcttc acctagatcc
     9241 ttttcacgta gaaagccagt ccgcagaaac ggtgctgacc ccggatgaat gtcagctact
     9301 gggctatctg gacaagggaa aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc
     9361 ttacatggcg atagctagac tgggcggttt tatggacagc aagcgaaccg gaattgccag
     9421 ctggggcgcc ctctggtaag gttgggaagc cctgcaaagt aaactggatg gctttcttgc
     9481 cgccaaggat ctgatggcgc aggggatcaa gctctgatca agagacagga tgaggatcgt
     9541 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     9601 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     9661 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     9721 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     9781 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     9841 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     9901 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     9961 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
    10021 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
    10081 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
    10141 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
    10201 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
    10261 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
    10321 ttcttgacga gttcttctga attattaacg cttacaattt cctgatgcgg tattttctcc
    10381 ttacgcatct gtgcggtatt tcacaccgca tcaggtggca cttttcgggg aaatgtgcgc
    10441 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagatta
    10501 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
    10561 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
    10621 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
    10681 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
    10741 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
    10801 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
    10861 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
    10921 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
    10981 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
    11041 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
    11101 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
    11161 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
    11221 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
    11281 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
    11341 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
    11401 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
    11461 tttcaatatt attgaagcat ttatcagggt tattgtctca tgaccaaaat cccttaacgt
    11521 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
    11581 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
    11641 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
    11701 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
    11761 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
    11821 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
    11881 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
    11941 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
    12001 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
    12061 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
    12121 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
    12181 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
    12241 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
    12301 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aag
//
