LOCUS       sequence-229324-       10535 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for FLPe recombinase-mediated cassette exchange in 
            master cell lines created with plasmid #112666. This vector allows 
            inducible expression of your protein of interest..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10535)
  AUTHORS   Ordovas L, Boon R, Pistoni M, Chen Y, Wolfs E, Guo W, Sambathkumar 
            R, Bobis-Wozowicz S, Helsen N, Vanhove J, Berckmans P, Cai Q, 
            Vanuytsel K, Eggermont K, Vanslembrouck V, Schmidt BZ, Raitano S, 
            Van Den Bosch L, Nahmias Y, Cathomen T, Struys T, Verfaillie CM
  TITLE     Efficient Recombinase-Mediated Cassette Exchange in hPSCs to Study 
            the Hepatocyte Lineage Reveals AAVS1 Locus-Mediated Transgene 
            Inhibition.
  JOURNAL   Stem Cell Reports. 2015 Nov 10;5(5):918-931. doi: 
            10.1016/j.stemcr.2015.09.004. Epub 2015 Oct 8.
  PUBMED    26455413
REFERENCE   2  (bases 1 to 10535)
  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.2015.09.004"; journalName: "Stem Cell Reports"; 
            date: "2015-11-10- 10"; volume: "5"; issue: "5"; pages: "918-931"
COMMENT     Sequence Label: pZ P TetON-3XFLAG-tdT CAGG-m2rtTA v2
FEATURES             Location/Qualifiers
     source          1..10535
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..10
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      94..103
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             100..1530
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     primer_bind     complement(1550..1569)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1594..1641
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     polyA_signal    1896..2007
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(2161..2760)
                     /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(2244..2264)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     2741..2760
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(2770..2823)
                     /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"
     misc_feature    2844..2869
                     /label=SA
                     /note="splice acceptor site"
     protein_bind    complement(2873..2906)
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     misc_feature    2913..3716
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     primer_bind     complement(3730..3749)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3731..3749)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3759..3776)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3759..3775)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3768..3790)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3917..4372
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4004..4023)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4214..4235
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4398..4502
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4503..4571,4572..5363)
                     /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     complement(4721..4740)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5534..6122
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6023..6042
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6276..6293
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6410..6431
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6446..6451,6452..6469,6470..6476)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6484..6500
                     /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     6489..6511
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6508..6524
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6508..6524
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6543..6563
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6545..6563
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    6593..7429
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     protein_bind    complement(7439..7472)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     polyA_signal    7479..7703
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(join(7717..7725,7726..7761,7762..7764,7765..7800,
                     7801..7803,7804..7839,7840..7845,7846..8463))
                     /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"
     primer_bind     complement(8491..8510)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          8518..9534
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(8572..8594)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(9535..9812)
                     /label=chicken beta-actin promoter
     enhancer        9817..10200
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    10261..10279
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10296..10314
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10332..10350
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10368..10386
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10403..10421
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10439..10457
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        10469..10507
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     10504..10528
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 gccaccatgg actacaaaga ccatgacggt gattataaag atcatgatat tgattacaag
       61 gatgacgatg acgaccggtg gagatatcca tatgccacca tggtgagcaa gggcgaggag
      121 gtcatcaaag agttcatgcg cttcaaggtg cgcatggagg gctccatgaa cggccacgag
      181 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
      241 aaggtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc ccagttcatg
      301 tacggctcca aggcgtacgt gaagcacccc gccgacatcc ccgattacaa gaagctgtcc
      361 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg tctggtgacc
      421 gtgacccagg actcctccct gcaggacggc acgctgatct acaaggtgaa gatgcgcggc
      481 accaacttcc cccccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
      541 accgagcgcc tgtacccccg cgacggcgtg ctgaagggcg agatccacca ggccctgaag
      601 ctgaaggacg gcggccacta cctggtggag ttcaagacca tctacatggc caagaagccc
      661 gtgcaactgc ccggctacta ctacgtggac accaagctgg acatcacctc ccacaacgag
      721 gactacacca tcgtggaaca gtacgagcgc tccgagggcc gccaccacct gttcctgggg
      781 catggcaccg gcagcaccgg cagcggcagc tccggcaccg cctcctccga ggacaacaac
      841 atggccgtca tcaaagagtt catgcgcttc aaggtgcgca tggagggctc catgaacggc
      901 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
      961 aagctgaagg tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag
     1021 ttcatgtacg gctccaaggc gtacgtgaag caccccgccg acatccccga ttacaagaag
     1081 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggtctg
     1141 gtgaccgtga cccaggactc ctccctgcag gacggcacgc tgatctacaa ggtgaagatg
     1201 cgcggcacca acttcccccc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     1261 gcctccaccg agcgcctgta cccccgcgac ggcgtgctga agggcgagat ccaccaggcc
     1321 ctgaagctga aggacggcgg ccactacctg gtggagttca agaccatcta catggccaag
     1381 aagcccgtgc aactgcccgg ctactactac gtggacacca agctggacat cacctcccac
     1441 aacgaggact acaccatcgt ggaacagtac gagcgctccg agggccgcca ccacctgttc
     1501 ctgtacggca tggacgagct gtacaagtaa acgcgtgggg gaggctaact gaaacacgga
     1561 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     1621 acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     1681 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     1741 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     1801 ccatagctcg agtgctattg tcttcccaat cctccccctt gctgtcctgc cccaccccac
     1861 cccccagaat agaatgacac ctactcagac aatgcgatgc aatttcctca ttttattagg
     1921 aaaggacagt gggagtggca ccttccaggg tcaaggaagg cacgggggag gggcaaacaa
     1981 cagatggctg gcaactagaa ggcacagtcg aggctgatca gcgggtttaa acgggccctc
     2041 tagagtcggt gggcctcggg ggcgggtgcg gggtcggcgg ggccgccccg ggtggcttcg
     2101 gtcggagcca tggggtcgtg cgctcctttc ggtcgggcgc tgcgggtcgt ggggcgggcg
     2161 tcaggcaccg ggcttgcggg tcatgcacca ggtgcgcggt ccttcgggca cctcgacgtc
     2221 ggcggtgacg gtgaagccga gccgctcgta gaaggggagg ttgcggggcg cggaggtctc
     2281 caggaaggcg ggcaccccgg cgcgctcggc cgcctccact ccggggagca cgacggcgct
     2341 gcccagaccc ttgccctggt ggtcgggcga gacgccgacg gtggccagga accacgcggg
     2401 ctccttgggc cggtgcggcg ccaggaggcc ttccatctgt tgctgcgcgg ccagccggga
     2461 accgctcaac tcggccatgc gcgggccgat ctcggcgaac accgcccccg cttcgacgct
     2521 ctccggcgtg gtccagaccg ccaccgcggc gccgtcgtcc gcgacccaca ccttgccgat
     2581 gtcgagcccg acgcgcgtga ggaagagttc ttgcagctcg gtgacccgct cgatgtggcg
     2641 gtccgggtcg acggtgtggc gcgtggcggg gtagtcggcg aacgcggcgg cgagggtgcg
     2701 tacggcccgg gggacgtcgt cgcgggtggc gaggcgcacc gtgggcttgt actcggtcat
     2761 ttcgagccta gggccgggat tctcctccac gtcaccgcat gttagaagac ttcctctgcc
     2821 ctctccgctg ccagatctga ggccctgtgg gaggaagaga agaggtcagg ccgaagttcc
     2881 tatactattt gaagaatagg aacttcaagc ttgccccact gtggggtgga ggggacagat
     2941 aaaagtaccc agaaccagag ccacattaac cggccctggg aatataaggt ggtcccagct
     3001 cggggacaca ggatccctgg aggcagcaaa catgctgtcc tgaagtggac ataggggccc
     3061 gggttggagg aagaagacta gctgagctct cggacccctg gaagatgcca tgacaggggg
     3121 ctggaagagc tagcacagac tagagaggta aggggggtag gggagctgcc caaatgaaag
     3181 gagtgagagg tgacccgaat ccacaggaga acggggtgtc caggcaaaga aagcaagagg
     3241 atggagaggt ggctaaagcc agggagacgg ggtactttgg ggttgtccag aaaaacggtg
     3301 atgatgcagg cctacaagaa ggggaggcgg gacgcaaggg agacatccgt cggagaaggc
     3361 catcctaaga aacgagagat ggcacaggcc ccagaaggag aaggaaaagg gaacccagcg
     3421 agtgaagacg gcatggggtt gggtgaggga ggagagatgc ccggagagga cccagacacg
     3481 gggaggatcc gctcagagga catcacgtgg tgcagcgccg agaaggaagt gctccggaaa
     3541 gagcatcctt gggcagcaac acagcagaga gcaaggggaa gagggagtgg aggaagacgg
     3601 aacctgaagg aggcggcagg gaaggatctg ggccagccgt agaggtgacc caggccacaa
     3661 gctgcagaca gaaagcggca caggcccagg ggagagaatg ctggtcagag aaagcaggta
     3721 cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt
     3781 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     3841 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     3901 ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3961 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     4021 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4081 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     4141 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4201 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4261 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4321 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca
     4381 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4441 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4501 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4561 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4621 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4681 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4741 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4801 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4861 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4921 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4981 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5041 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5101 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5161 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5221 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5281 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5341 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5401 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5461 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5521 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5581 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5641 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5701 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5761 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5821 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5881 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5941 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6001 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6061 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6121 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6181 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     6241 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     6301 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     6361 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     6421 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     6481 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     6541 gcgcaattaa ccctcactaa agggaacaaa agctggagct ccaccgcggt ggagagcaga
     6601 gccaggaacc cctgtaggga aggggcagga gagccagggg catgagatgg tggacgagga
     6661 agggggacag ggaagcctga gcgcctctcc tgggcttgcc aaggactcaa acccagaagc
     6721 ccagagcagg gccttaggga agcgggaccc tgctctgggc ggaggaatat gtcccagata
     6781 gcactgggga ctctttaagg aaagaaggat ggagaaagag aaagggagta gaggcggcca
     6841 cgacctggtg aacacctagg acgcaccatt ctcacaaagg gagttttcca cacggacacc
     6901 cccctcctca ccacagccct gccaggacgg ggctggctac tggccttatc tcacaggtaa
     6961 aactgacgca cggaggaaca atataaattg gggactagaa aggtgaagag ccaaagttag
     7021 aactcaggac caacttattc tgattttgtt tttccaaact gcttctcctc ttgggaagtg
     7081 taaggaagct gcagcaccag gatcagtgaa acgcaccaga cagccgcgtc agagcagctc
     7141 aggttctggg agagggtagc gcagggtggc cactgagaac cgggcaggtc acgcatcccc
     7201 cccttccctc ccaccccctg ccaagctctc cctcccagga tcctctctgg ctccatcgta
     7261 agcaaacctt agaggttctg gcaaggagag agatggctcc aggaaatggg ggtgtgtcac
     7321 cagataagga atctgcctaa caggaggtgg gggttagacc caatatcagg agactaggaa
     7381 ggaggaggcc taaggatggg gcttttctgt caccaatcct gtccctagta gcggccgcga
     7441 agttcctata ctttctagag aataggaact tcttcgaacc atagagccca ccgcatcccc
     7501 agcatgcctg ctattgtctt cccaatcctc ccccttgctg tcctgcccca ccccaccccc
     7561 cagaatagaa tgacacctac tcagacaatg cgatgcaatt tcctcatttt attaggaaag
     7621 gacagtggga gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga
     7681 tggctggcaa ctagaaggca cagtcgaggc actagtttac ccggggagca tgtcaaggtc
     7741 aaaatcgtca agagcgtcag caggcagcat atcaaggtca aagtcgtcaa gggcatcggc
     7801 tgggagcatg tctaagtcaa aatcgtcaag ggcgtcggcc ggcccgccgc tttcgcactt
     7861 tagctgtttc tccaggccac atatgattag ttccaggccg aaaaggaagg caggttcggc
     7921 tccctgccgg tcgaacagct caattgcttg tctcagaagt gggggcatag aatcggtggt
     7981 aggtgtctct ctttcctctt ttgctacttg atgctcctgt tcctccaata cgcagcccag
     8041 tgtaaagtgg cccacggcgg acagagcgta cagtgcgttc tccagggaga agccttgctg
     8101 acacaggaac gcgagctgat tttccagggt ttcgtactgt ttctctgttg ggcgggtgcc
     8161 gagatgcact ttagccccgt cgcgatgtga gaggagagca cagcggaatg acttggcgtt
     8221 gttccgcaga aagtcttgcc atgactcgcc ttccaggggg cagaagtggg tatgatgcct
     8281 gtccagcatc tcgattggca gggcatcgag cagggcccgc ttgttcttca cgtgccagta
     8341 cagggtaggc tgctcaactc ccagcttttg agcgagtttc cttgtcgtca ggccttcgat
     8401 accgactcca ttgagtaatt ccagagcgcc gtttatgact ttgctcttgt ccagtctaga
     8461 catgaattct ttgccaaaat gatgagacag cacaataacc agcacgttgc ccaggagctg
     8521 taggaaaaag aagaaggcat gaacatggtt agcagaggct ctagagccgc cggtcacacg
     8581 ccagaagccg aaccccgccc tgccccgtcc cccccgaagg cagccgtccc cccgcggaca
     8641 gccccgaggc tggagaggga gaaggggacg gcggcgcggc gacgcacgaa ggccctcccc
     8701 gcccatttcc ttcctgccgg cgccgcaccg cttcgccccg cgcccgctag agggggtgcg
     8761 gcggcgcctc ccagatttcg gctccgcaca gatttgggac aaaggaagtc cctgcgccct
     8821 ctcgcacgat taccataaaa ggcaatggct gcggctcgcc gcgcctcgac agccgccggc
     8881 gctccggggg ccgccgcgcc cctcccccga gccctccccg gcccgaggcg gccccgcccc
     8941 gcccggcacc cccacctgcc gccacccccc gcccggcacg gcgagccccg cgccacgccc
     9001 cgtacggagc cccgcacccg aagccgggcc gtgctcagca actcggggag gggggtgcag
     9061 ggggggttgc agcccgaccg acgcgcccac accccctgct caccccccca cgcacacacc
     9121 ccgcacgcag cctttgttcc cctcgcagcc ccccccgcac cgcggggcac cgcccccggc
     9181 cgcgctcccc tcgcgcacac tgcggagcgc acaaagcccc gcgccgcgcc cgcagcgctc
     9241 acagccgccg ggcagcgcgg agccgcacgc ggcgctcccc acgcacacac acacgcacgc
     9301 accccccgag ccgctccccc cgcacaaagg gccctcccgg agcccctcaa ggctttcacg
     9361 cagccacaga aaagaaacaa gccgtcatta aaccaagcgc taattacagc ccggaggaga
     9421 agggccgtcc cgcccgctca cctgtgggag taacgcggtc agtcagagcc ggggcgggcg
     9481 gcgcgaggcg gcggcggagc ggggcacggg gcgaaggcag cgcgcagcga ctcccgcccg
     9541 ccgcgcgctt cgctttttat agggccgccg ccgccgccgc ctcgccataa aaggaaactt
     9601 tcggagcgcg ccgctctgat tggctgccgc cgcacctctc cgcctcgccc cgccccgccc
     9661 ctcgccccgc cccgccccgc ctggcgcgcg cccccccccc ccccccgccc ccatcgctgc
     9721 acaaaataat taaaaaataa ataaatacaa aattgggggt ggggaggggg gggagatggg
     9781 gagagtgaag cagaacgtgg ggctcacctc gaccatcatg gtaatagcga tgactaatac
     9841 gtagatgtac tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc
     9901 gggccattta ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg
     9961 tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc
    10021 tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg
    10081 cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct
    10141 atgaactaat gaccccgtaa ttgattacta ttaataacta gctagtcaat aatcaatgtc
    10201 gacgatgatg ctgtacactc gaaacttttt tcgaaggtcc gaggttctag acgagtttac
    10261 tccctatcag tgatagagaa cgatgtcgag tttactccct atcagtgata gagaacgtat
    10321 gtcgagttta ctccctatca gtgatagaga acgtatgtcg agtttactcc ctatcagtga
    10381 tagagaacgt atgtcgagtt tatccctatc agtgatagag aacgtatgtc gagtttactc
    10441 cctatcagtg atagagaacg tatgtcgagg taggcgtgta cggtgggagg cctatataag
    10501 cagagctcgt ttagtgaacc gtcagatcgc ttaag
//
