LOCUS       sequence-247988-        9985 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Allows for doxycycline-inducible gene expression from the murine 
            ROSA26 safe harbor locus upon CRISPR/Cas9-mediated genomic insertion
            and stable selection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9985)
  AUTHORS   Routh ED, Pullikuth AK, Jin G, Chifman J, Chou JW, D'Agostino RB Jr,
            Seino KI, Wada H, Print CG, Zhang W, Lu Y, Miller LD
  TITLE     Transcriptomic Features of T Cell-Barren Tumors Are Conserved Across
            Diverse Tumor Types.
  JOURNAL   Front Immunol. 2020 Feb 13;11:57. doi: 10.3389/fimmu.2020.00057. 
            eCollection 2020.
  PUBMED    32117236
REFERENCE   2  (bases 1 to 9985)
  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: "Front 
            Immunol."; date: "2020-02-13"; volume: "11:57. doi"; pages: " 
            10.3389/fimmu.2020.00057. eCollection 2020"
COMMENT     Sequence Label: pR26
FEATURES             Location/Qualifiers
     source          1..9985
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..794
                     /label=Rosa26 left arm
                     /note="left homology arm from intron 1 of the mouse Rosa26 
                     gene (Soriano, 1999)"
     misc_feature    join(813..878,879..916)
                     /label=Ad SA
                     /note="adenovirus major late transcript splice acceptor 
                     (Friedrich and Soriano, 1991)"
     polyA_signal    931..979
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        1004..1318
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    1012..1030
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1048..1066
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1083..1101
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1119..1137
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1155..1173
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1190..1208
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1226..1244
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1291..1315
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     polyA_signal    1419..1467
                     /note="synthetic polyadenylation signal"
     promoter        1481..1758
                     /label=chicken beta-actin promoter
     intron          2061..2633
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     2641..2660
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     promoter        2688..2706
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(2730..3347,3348..3353,3354..3389,3390..3392,3393..3428,
                     3429..3431,3432..3467,3468..3476)
                     /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
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGAR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     polyA_signal    3479..3704
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4747..4766
                     /label=hUBCpro-F
                     /note="Human Ubiquitin C (UbC) promoter, forward primer"
     CDS             4925..5524
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVESAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4925..4944)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5421..5441
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5525..5646
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5562..5581)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5616..5635
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    5703..6513
                     /label=Rosa26 right arm
                     /note="right homology arm from intron 1 of the mouse Rosa26
                     gene (Soriano, 1999)"
     promoter        complement(6588..6606)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6613..6630)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6613..6629)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6622..6644)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             6767..7069
                     /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     6948..6967
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             7418..8212
                     /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             7418..8212
                     /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(7472..7491)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8082..8101
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     regulatory      8412..8421
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8418..8792
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     rep_origin      8930..9518
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9419..9438
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9672..9689
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9806..9827
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9842..9847,9848..9865,9866..9872)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9880..9896
                     /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     9885..9907
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9904..9920
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9904..9920
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        9938..9956
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     9938..9955
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 cagttaacgg cagccggagt gcgcagccgc cggcagcctc gctctgccca ctgggtgggg
       61 cgggaggtag gtggggtgag gcgagctgga cgtgcgggcg cggtcggcct ctggcggggc
      121 gggggggggg agggagggtc agcgaaagta gctcgcgcgc gagcggccgc ccaccctccc
      181 cttcctctgg gggagtcgtt ttacccgccg ccggccgggc ctcgtcgtct gattggctct
      241 cggggcccag aaaactggcc cttgccattg gctcgtgttc gtgcaagttg agtccatccg
      301 ccggccagcg ggggcggcga ggaggcgctc ccaggttccg gccctcccct cggccccgcg
      361 ccgcagagtc tggccgcgcg cccctgcgca acgtggcagg aagcgcgcgc tgggggcggg
      421 gacgggcagt agggctgagc ggctgcgggg cgggtgcaag cacgtttccg acttgagttg
      481 cctcaagagg ggcgtgctga gccagacctc catcgcgcac tccggggagt ggagggaagg
      541 agcgagggct cagttgggct gttttggagg caggaagcac ttgctctccc aaagtcgctc
      601 tgagttgtta tcagtaaggg agctgcagtg gagtaggcgg ggagaaggcc gcacccttct
      661 ccggaggggg gaggggagtg ttgcaatacc tttctgggag ttctctgctg cctcctggct
      721 tctgaggacc gccctgggcc tgggagaatc ccttccccct cttccctcgt gatctgcaac
      781 tccagtcttt ctagaatcga tctcgagaac tgtagggcgc agtagtccag ggtttccttg
      841 atgatgtcat acttatcctg tccctttttt ttccacagct cgcggttgag gacaaactct
      901 tcgcggtctt tccagtggtt aattagggat cggcaataaa aagacagaat aaaacgcacg
      961 ggtgttgggt cgtttgttca tcgatgccct ttcgtcttca ttcgagttta ctccctatca
     1021 gtgatagaga acgtatgtcg agtttactcc ctatcagtga tagagaacga tgtcgagttt
     1081 actccctatc agtgatagag aacgtatgtc gagtttactc cctatcagtg atagagaacg
     1141 tatgtcgagt ttactcccta tcagtgatag agaacgtatg tcgagtttat ccctatcagt
     1201 gatagagaac gtatgtcgag tttactccct atcagtgata gagaacgtat gtcgaggtag
     1261 gcgtgtacgg tgggaggcct atataagcag agctcgttta gtgaaccgtc agatcgcctg
     1321 gagaattcga gctcggtacc cggggatgct agctctaggt taattaagcc cgggccctgc
     1381 aggtaggatg aatttaaatg tttaaacacg cgttaggaaa taaaagatct ttattttcat
     1441 tagatctgtg tgttggtttt ttgtgtgccg cgtcctagag tcgaggtgag ccccacgttc
     1501 tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt
     1561 taattatttt gtgcagcgat gggggcgcgg gggggggggg ggcgcgcgcc gggggggggg
     1621 ggggggggcg gggggggggg cggggcgagg cggagaggtg cggcggcagc caatcagagc
     1681 ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag
     1741 cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc
     1801 gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca ggtgagcggg
     1861 cgggacggcc cttctcctcc gggctgtaat tagcgcttgg tttaatgacg gcttgtttct
     1921 tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg atcttctagg gcctctgagc
     1981 tattccagaa gtagtgaaga ggcttttttg gaggcctagg cttttgcaaa aagctccgga
     2041 tcgatcctga gaacttcagg gtgagtttgg ggacccttga ttgttctttc tttttcgcta
     2101 ttgtaaaatt catgttatat ggagggggca aagttttcag ggtgttgttt agaatgggaa
     2161 gatgtccctt gtatcaccat ggaccctcat gataattttg tttctttcac tttctactct
     2221 gttgacaacc attgtctcct cttattttct tttcattttc tgtaactttt tcgttaaact
     2281 ttagcttgca tttgtaacga atttttaaat tcacttttgt ttatttgtca gattgtaagt
     2341 actttctcta atcacttttt tttcaaggca atcagggtat attatattgt acttcagcac
     2401 agttttagag aacaattgtt ataattaaat gataaggtag aatatttctg catataaatt
     2461 ctggctggcg tggaaatatt cttattggta gaaacaacta catcctggtc atcatcctgc
     2521 ctttctcttt atggttacaa tgatatacac tgtttgagat gaggataaaa tactctgagt
     2581 ccaaaccggg cccctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg
     2641 gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattgtaa tacgactcac
     2701 tatagggcga attgatcgga tccgccacca tgtccagact ggacaagagc aaagtcataa
     2761 acggcgctct ggaattactc aatggagtcg gtatcgaagg cctgacgaca aggaaactcg
     2821 ctcaaaagct gggagttgag cagcctaccc tgtactggca cgtgaagaac aagcgggccc
     2881 tgctcgatgc cctgccaatc gagatgctgg acaggcatca tacccacttc tgccccctgg
     2941 aaggcgagtc atggcaagac tttctgcgga acaacgccaa gtcattccgc tgtgctctcc
     3001 tctcacatcg cgacggggct aaagtgcatc tcggcgcccg cccaacagag aaacagtacg
     3061 aaaccctgga aaatcagctc gcgttcctgt gtcagcaagg cttctccctg gagaacgcac
     3121 tgtacgctct gtccgccgtg ggccacttta cactgggctg cgtattggag gaacaggagc
     3181 atcaagtagc aaaagaggaa agagagacac ctaccaccga ttctatgccc ccacttctga
     3241 gacaagcaat tgagctgttc gaccggcagg gagccgaacc tgccttcctt ttcggcctgg
     3301 aactaatcat atgtggcctg gagaaacagc taaagtgcga aagcggcggg ccggccgacg
     3361 cccttgacga ttttgactta gacatgctcc cagccgatgc ccttgacgac tttgaccttg
     3421 atatgctgcc tgctgacgct cttgacgatt ttgaccttga catgctcccc gggtaaggct
     3481 gtgcctttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     3541 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     3601 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     3661 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcgcgcc gggttttggc
     3721 gcctcccgcg ggcgcccccc tcctcacggc gagcgctgcc acgtcagacg aagggcgcag
     3781 cgagcgtcct gatccttccg cccggacgct caggacagcg gcccgctgct cataagactc
     3841 ggccttagaa ccccagtatc agcagaagga cattttagga cgggacttgg gtgactctag
     3901 ggcactggtt ttctttccag agagcggaac aggcgaggaa aagtagtccc ttctcggcga
     3961 ttctgcggag ggatctccgt ggggcggtga acgccgatga ttatataagg acgcgccggg
     4021 tgtggcacag ctagttccgt cgcagccggg atttgggtcg cggttcttgt ttgtggatcg
     4081 ctgtgatcgt cacttggtga gtagcgggct gctgggctgg ccggggcttt cgtggccgcc
     4141 gggccgctcg gtgggacgga agcgtgtgga gagaccgcca agggctgtag tctgggtccg
     4201 cgagcaaggt tgccctgaac tgggggttgg ggggagcgca gcaaaatggc ggctgttccc
     4261 gagtcttgaa tggaagacgc ttgtgaggcg ggctgtgagg tcgttgaaac aaggtggggg
     4321 gcatggtggg cggcaagaac ccaaggtctt gaggccttcg ctaatgcggg aaagctctta
     4381 ttcgggtgag atgggctggg gcaccatctg gggaccctga cgtgaagttt gtcactgact
     4441 ggagaactcg gtttgtcgtc tgttgcgggg gcggcagtta tggcggtgcc gttgggcagt
     4501 gcacccgtac ctttgggagc gcgcgccctc gtcgtgtcgt gacgtcaccc gttctgttgg
     4561 cttataatgc agggtggggc cacctgccgg taggtgtgcg gtaggctttt ctccgtcgca
     4621 ggacgcaggg ttcgggccta gggtaggctc tcctgaatcg acaggcgccg gacctctggt
     4681 gaggggaggg ataagtgagg cgtcagtttc tttggtcggt tttatgtacc tatcttctta
     4741 agtagctgaa gctccggttt tgaactatgc gctcggggtt ggcgagtgtg ttttgtgaag
     4801 ttttttaggc accttttgaa atgtaatcat ttgggtcaat atgtaatttt cagtgttaga
     4861 ctagtaaatt gtccgctaaa ttctggccgt ttttggcttt tttgttagac caagtttggc
     4921 caccatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc
     4981 cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc
     5041 ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct
     5101 cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc
     5161 ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag
     5221 cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa
     5281 ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct
     5341 gggcagcgcc gtcgtgctcc ccggagtgga gtcggccgag cgcgccgggg tgcccgcctt
     5401 cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac
     5461 cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc
     5521 ctgaaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     5581 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     5641 atcttagggt accgagctcg gatccactag taacggccgc cagtgtgctg gaattcgccc
     5701 ttagatgggc gggagtcttc tgggcaggct taaaggctaa cctggtgtgt gggcgttgtc
     5761 ctgcagggga attgaacagg tgtaaaattg gagggacaag acttcccaca gattttcggt
     5821 tttgtcggga agttttttaa taggggcaaa taaggaaaat gggaggatag gtagtcatct
     5881 ggggttttat gcagcaaaac tacaggttat tattgcttgt gatccgcctc ggagtatttt
     5941 ccatcgaggt agattaaaga catgctcacc cgagttttat actctcctgc ttgagatcct
     6001 tactacagta tgaaattaca gtgtcgcgag ttagactatg taagcagaat tttaatcatt
     6061 tttaaagagc ccagtacttc atatccattt ctcccgctcc ttctgcagcc ttatcaaaag
     6121 gtattttaga acactcattt tagccccatt ttcatttatt atactggctt atccaacccc
     6181 tagacagagc attggcattt tccctttcct gatcttagaa gtctgatgac tcatgaaacc
     6241 agacagatta gttacataca ccacaaatcg aggctgtagc tggggcctca acactgcagt
     6301 tcttttataa ctccttagta cactttttgt tgatcctttg ccttgatcct taattttcag
     6361 tgtctatcac ctctcccgtc aggtggtgtt ccacatttgg gcctattctc agtccaggga
     6421 gttttacaac aatagatgta ttgagaatcc aacctaaagc ttaactttcc actcccatga
     6481 atgcctctct cctttttctc catttataaa ctgaagggcg aattctgcag atatccatca
     6541 cactggcggc cgctcgatcg agcatgcatc tagagggccc aattcgccct atagtgagtc
     6601 gtattacaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     6661 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     6721 ccgcaccgat cgcccttccc aacagttgcg cagcctatac gtacggcagt ttaaggttta
     6781 cacctataaa agagagagcc gttatcgtct gtttgtggat gtacagagtg atattattga
     6841 cacgccgggg cgacggatgg tgatccccct ggccagtgca cgtctgctgt cagataaagt
     6901 ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa agctggcgca tgatgaccac
     6961 cgatatggcc agtgtgccgg tctccgttat cggggaagaa gtggctgatc tcagccaccg
     7021 cgaaaatgac atcaaaaacg ccattaacct gatgttctgg ggaatataaa tgtcaggcat
     7081 gagattatca aaaaggatct tcacctagat ccttttcacg tagaaagcca gtccgcagaa
     7141 acggtgctga ccccggatga atgtcagcta ctgggctatc tggacaaggg aaaacgcaag
     7201 cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg cgatagctag actgggcggt
     7261 tttatggaca gcaagcgaac cggaattgcc agctggggcg ccctctggta aggttgggaa
     7321 gccctgcaaa gtaaactgga tggctttctt gccgccaagg atctgatggc gcaggggatc
     7381 aagctctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     7441 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     7501 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     7561 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     7621 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     7681 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     7741 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     7801 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     7861 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     7921 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     7981 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     8041 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     8101 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     8161 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gaattattaa
     8221 cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     8281 catcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     8341 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatagc
     8401 acgtgaggag ggccaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg
     8461 acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg
     8521 aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg
     8581 accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt
     8641 acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga
     8701 ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact
     8761 gcgtgcactt cgtggccgag gagcaggact gacacgtgct aaaacttcat ttttaattta
     8821 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     8881 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     8941 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     9001 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     9061 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     9121 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     9181 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     9241 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     9301 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     9361 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     9421 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     9481 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     9541 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     9601 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     9661 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     9721 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     9781 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     9841 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     9901 acacaggaaa cagctatgac catgattacg ccaagctatt taggtgacac tatagaatac
     9961 tcaagctatg catcaagctt ggtac
//
