LOCUS       sequence-229038-       13238 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 and contains cHS4 insulators..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13238)
  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 13238)
  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 (cHS4)X4 TetON-3XFLAG-tdT CAGG-m2rtTA v2
FEATURES             Location/Qualifiers
     source          1..13238
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..19
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    37..55
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    73..91
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    108..126
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    144..162
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        174..212
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     209..233
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      241..250
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      334..343
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             340..1770
                     /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(1790..1809)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1834..1881
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     polyA_signal    3485..3596
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(3750..4349)
                     /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(3833..3853)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     4330..4349
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(4359..4412)
                     /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    4433..4458
                     /label=SA
                     /note="splice acceptor site"
     protein_bind    complement(4462..4495)
                     /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    4502..5305
                     /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(5319..5338)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5320..5338)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5348..5365)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5348..5364)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5357..5379)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5506..5961
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5593..5612)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5803..5824
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5987..6091
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6092..6160,6161..6952)
                     /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(6310..6329)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7123..7711
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7612..7631
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7865..7882
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7999..8020
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8035..8040,8041..8058,8059..8065)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8073..8089
                     /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     8078..8100
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8097..8113
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8097..8113
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8132..8152
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8134..8152
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    8182..9018
                     /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(9028..9061)
                     /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    10422..10646
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(join(10660..10668,10669..10704,10705..10707,
                     10708..10743,10744..10746,10747..10782,10783..10788,
                     10789..11406))
                     /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(11434..11453)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          11461..12477
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(11515..11537)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(12478..12755)
                     /label=chicken beta-actin promoter
     enhancer        12760..13143
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    13204..13222
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat agagaacgta
       61 tgtcgagttt actccctatc agtgatagag aacgtatgtc gagtttatcc ctatcagtga
      121 tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgtatgt cgaggtaggc
      181 gtgtacggtg ggaggcctat ataagcagag ctcgtttagt gaaccgtcag atcgcttaag
      241 gccaccatgg actacaaaga ccatgacggt gattataaag atcatgatat tgattacaag
      301 gatgacgatg acgaccggtg gagatatcca tatgccacca tggtgagcaa gggcgaggag
      361 gtcatcaaag agttcatgcg cttcaaggtg cgcatggagg gctccatgaa cggccacgag
      421 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
      481 aaggtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc ccagttcatg
      541 tacggctcca aggcgtacgt gaagcacccc gccgacatcc ccgattacaa gaagctgtcc
      601 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg tctggtgacc
      661 gtgacccagg actcctccct gcaggacggc acgctgatct acaaggtgaa gatgcgcggc
      721 accaacttcc cccccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
      781 accgagcgcc tgtacccccg cgacggcgtg ctgaagggcg agatccacca ggccctgaag
      841 ctgaaggacg gcggccacta cctggtggag ttcaagacca tctacatggc caagaagccc
      901 gtgcaactgc ccggctacta ctacgtggac accaagctgg acatcacctc ccacaacgag
      961 gactacacca tcgtggaaca gtacgagcgc tccgagggcc gccaccacct gttcctgggg
     1021 catggcaccg gcagcaccgg cagcggcagc tccggcaccg cctcctccga ggacaacaac
     1081 atggccgtca tcaaagagtt catgcgcttc aaggtgcgca tggagggctc catgaacggc
     1141 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     1201 aagctgaagg tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag
     1261 ttcatgtacg gctccaaggc gtacgtgaag caccccgccg acatccccga ttacaagaag
     1321 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggtctg
     1381 gtgaccgtga cccaggactc ctccctgcag gacggcacgc tgatctacaa ggtgaagatg
     1441 cgcggcacca acttcccccc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     1501 gcctccaccg agcgcctgta cccccgcgac ggcgtgctga agggcgagat ccaccaggcc
     1561 ctgaagctga aggacggcgg ccactacctg gtggagttca agaccatcta catggccaag
     1621 aagcccgtgc aactgcccgg ctactactac gtggacacca agctggacat cacctcccac
     1681 aacgaggact acaccatcgt ggaacagtac gagcgctccg agggccgcca ccacctgttc
     1741 ctgtacggca tggacgagct gtacaagtaa acgcgtgggg gaggctaact gaaacacgga
     1801 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     1861 acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     1921 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     1981 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     2041 ccatagctcg agatcgaagc gcatttgatc gatctcactg actccgtcct ggagttggat
     2101 gagagataat ggccttacgt tgtgccaggg gagggtcggg ctggatttag caagatttac
     2161 cttctccaaa gagcggtgct gcagtggcac agctgcccac ggaggtgggg gggtcaccgt
     2221 ccctggaggt gatgaagaac tgtggggatg tggcactgag ggacatggcc agtgggcacg
     2281 gtgggtgggt tggggttggt cttggggatc ttggagggct tttccagcct tcatgatttg
     2341 acgattgtat gaacatctac atggcaattc tccagctgcc tgtcccagtc ctactgaccc
     2401 agctgtatct ctccaggcaa gctcttccac cccttctgct tgcatccaga caccatcaaa
     2461 catgcaggct cagacacaag cctaaagctt tttccccgta tccccccagg tgtctgcagg
     2521 ctcaaagagc agcgagaagc gttcagagga aagcgatccc gtgccacctt ccccgtgccc
     2581 gggctgtccc cgcacgctgc cggctcgggg atgcgggggg agcgccggac cggagcggag
     2641 ccccgggcgg ctcgctgctg ccccctagcg ggggagggac gtaattacat ccctgggggc
     2701 tttggggggg ggctgtcccc gtatgcatct cactgactcc gtcctggagt tggatgagag
     2761 ataatggcct tacgttgtgc caggggaggg tcgggctgga tttagcaaga tttaccttct
     2821 ccaaagagcg gtgctgcagt ggcacagctg cccacggagg tgggggggtc accgtccctg
     2881 gaggtgatga agaactgtgg ggatgtggca ctgagggaca tggccagtgg gcacggtggg
     2941 tgggttgggg ttggtcttgg ggatcttgga gggcttttcc agccttcatg atttgacgat
     3001 tgtatgaaca tctacatggc aattctccag ctgcctgtcc cagtcctact gacccagctg
     3061 tatctctcca ggcaagctct tccacccctt ctgcttgcat ccagacacca tcaaacatgc
     3121 aggctcagac acaagcctaa agctttttcc ccgtatcccc ccaggtgtct gcaggctcaa
     3181 agagcagcga gaagcgttca gaggaaagcg atcccgtgcc accttccccg tgcccgggct
     3241 gtccccgcac gctgccggct cggggatgcg gggggagcgc cggaccggag cggagccccg
     3301 ggcggctcgc tgctgccccc tagcggggga gggacgtaat tacatccctg ggggctttgg
     3361 gggggggctg tccccgtatc gatacgctta ctagtgtcga gtgctattgt cttcccaatc
     3421 ctcccccttg ctgtcctgcc ccaccccacc ccccagaata gaatgacacc tactcagaca
     3481 atgcgatgca atttcctcat tttattagga aaggacagtg ggagtggcac cttccagggt
     3541 caaggaaggc acgggggagg ggcaaacaac agatggctgg caactagaag gcacagtcga
     3601 ggctgatcag cgggtttaaa cgggccctct agagtcggtg ggcctcgggg gcgggtgcgg
     3661 ggtcggcggg gccgccccgg gtggcttcgg tcggagccat ggggtcgtgc gctcctttcg
     3721 gtcgggcgct gcgggtcgtg gggcgggcgt caggcaccgg gcttgcgggt catgcaccag
     3781 gtgcgcggtc cttcgggcac ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag
     3841 aaggggaggt tgcggggcgc ggaggtctcc aggaaggcgg gcaccccggc gcgctcggcc
     3901 gcctccactc cggggagcac gacggcgctg cccagaccct tgccctggtg gtcgggcgag
     3961 acgccgacgg tggccaggaa ccacgcgggc tccttgggcc ggtgcggcgc caggaggcct
     4021 tccatctgtt gctgcgcggc cagccgggaa ccgctcaact cggccatgcg cgggccgatc
     4081 tcggcgaaca ccgcccccgc ttcgacgctc tccggcgtgg tccagaccgc caccgcggcg
     4141 ccgtcgtccg cgacccacac cttgccgatg tcgagcccga cgcgcgtgag gaagagttct
     4201 tgcagctcgg tgacccgctc gatgtggcgg tccgggtcga cggtgtggcg cgtggcgggg
     4261 tagtcggcga acgcggcggc gagggtgcgt acggcccggg ggacgtcgtc gcgggtggcg
     4321 aggcgcaccg tgggcttgta ctcggtcatt tcgagcctag ggccgggatt ctcctccacg
     4381 tcaccgcatg ttagaagact tcctctgccc tctccgctgc cagatctgag gccctgtggg
     4441 aggaagagaa gaggtcaggc cgaagttcct atactatttg aagaatagga acttcaagct
     4501 tgccccactg tggggtggag gggacagata aaagtaccca gaaccagagc cacattaacc
     4561 ggccctggga atataaggtg gtcccagctc ggggacacag gatccctgga ggcagcaaac
     4621 atgctgtcct gaagtggaca taggggcccg ggttggagga agaagactag ctgagctctc
     4681 ggacccctgg aagatgccat gacagggggc tggaagagct agcacagact agagaggtaa
     4741 ggggggtagg ggagctgccc aaatgaaagg agtgagaggt gacccgaatc cacaggagaa
     4801 cggggtgtcc aggcaaagaa agcaagagga tggagaggtg gctaaagcca gggagacggg
     4861 gtactttggg gttgtccaga aaaacggtga tgatgcaggc ctacaagaag gggaggcggg
     4921 acgcaaggga gacatccgtc ggagaaggcc atcctaagaa acgagagatg gcacaggccc
     4981 cagaaggaga aggaaaaggg aacccagcga gtgaagacgg catggggttg ggtgagggag
     5041 gagagatgcc cggagaggac ccagacacgg ggaggatccg ctcagaggac atcacgtggt
     5101 gcagcgccga gaaggaagtg ctccggaaag agcatccttg ggcagcaaca cagcagagag
     5161 caaggggaag agggagtgga ggaagacgga acctgaagga ggcggcaggg aaggatctgg
     5221 gccagccgta gaggtgaccc aggccacaag ctgcagacag aaagcggcac aggcccaggg
     5281 gagagaatgc tggtcagaga aagcaggtac ccaattcgcc ctatagtgag tcgtattacg
     5341 cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
     5401 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
     5461 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg
     5521 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     5581 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     5641 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     5701 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     5761 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     5821 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     5881 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     5941 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
     6001 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     6061 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     6121 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     6181 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     6241 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     6301 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     6361 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     6421 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     6481 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     6541 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     6601 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     6661 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     6721 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6781 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     6841 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6901 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     6961 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     7021 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     7081 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     7141 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     7201 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     7261 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     7321 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     7381 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     7441 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     7501 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     7561 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     7621 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     7681 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     7741 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     7801 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     7861 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     7921 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     7981 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     8041 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     8101 aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa
     8161 gctggagctc caccgcggtg gagagcagag ccaggaaccc ctgtagggaa ggggcaggag
     8221 agccaggggc atgagatggt ggacgaggaa gggggacagg gaagcctgag cgcctctcct
     8281 gggcttgcca aggactcaaa cccagaagcc cagagcaggg ccttagggaa gcgggaccct
     8341 gctctgggcg gaggaatatg tcccagatag cactggggac tctttaagga aagaaggatg
     8401 gagaaagaga aagggagtag aggcggccac gacctggtga acacctagga cgcaccattc
     8461 tcacaaaggg agttttccac acggacaccc ccctcctcac cacagccctg ccaggacggg
     8521 gctggctact ggccttatct cacaggtaaa actgacgcac ggaggaacaa tataaattgg
     8581 ggactagaaa ggtgaagagc caaagttaga actcaggacc aacttattct gattttgttt
     8641 ttccaaactg cttctcctct tgggaagtgt aaggaagctg cagcaccagg atcagtgaaa
     8701 cgcaccagac agccgcgtca gagcagctca ggttctggga gagggtagcg cagggtggcc
     8761 actgagaacc gggcaggtca cgcatccccc ccttccctcc caccccctgc caagctctcc
     8821 ctcccaggat cctctctggc tccatcgtaa gcaaacctta gaggttctgg caaggagaga
     8881 gatggctcca ggaaatgggg gtgtgtcacc agataaggaa tctgcctaac aggaggtggg
     8941 ggttagaccc aatatcagga gactaggaag gaggaggcct aaggatgggg cttttctgtc
     9001 accaatcctg tccctagtag cggccgcgaa gttcctatac tttctagaga ataggaactt
     9061 cttcgatctc gacaagcgta tcgatacggg gacagccccc ccccaaagcc cccagggatg
     9121 taattacgtc cctcccccgc tagggggcag cagcgagccg cccggggctc cgctccggtc
     9181 cggcgctccc cccgcatccc cgagccggca gcgtgcgggg acagcccggg cacggggaag
     9241 gtggcacggg atcgctttcc tctgaacgct tctcgctgct ctttgagcct gcagacacct
     9301 ggggggatac ggggaaaaag ctttaggctt gtgtctgagc ctgcatgttt gatggtgtct
     9361 ggatgcaagc agaaggggtg gaagagcttg cctggagaga tacagctggg tcagtaggac
     9421 tgggacaggc agctggagaa ttgccatgta gatgttcata caatcgtcaa atcatgaagg
     9481 ctggaaaagc cctccaagat ccccaagacc aaccccaacc cacccaccgt gcccactggc
     9541 catgtccctc agtgccacat ccccacagtt cttcatcacc tccagggacg gtgacccccc
     9601 cacctccgtg ggcagctgtg ccactgcagc accgctcttt ggagaaggta aatcttgcta
     9661 aatccagccc gaccctcccc tggcacaacg taaggccatt atctctcatc caactccagg
     9721 acggagtcag tgagatgcat acggggacag ccccccccca aagcccccag ggatgtaatt
     9781 acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc cggtccggcg
     9841 ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg ggaaggtggc
     9901 acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga cacctggggg
     9961 gatacgggga aaaagcttta ggcttgtgtc tgagcctgca tgtttgatgg tgtctggatg
    10021 caagcagaag gggtggaaga gcttgcctgg agagatacag ctgggtcagt aggactggga
    10081 caggcagctg gagaattgcc atgtagatgt tcatacaatc gtcaaatcat gaaggctgga
    10141 aaagccctcc aagatcccca agaccaaccc caacccaccc accgtgccca ctggccatgt
    10201 ccctcagtgc cacatcccca cagttcttca tcacctccag ggacggtgac ccccccacct
    10261 ccgtgggcag ctgtgccact gcagcaccgc tctttggaga aggtaaatct tgctaaatcc
    10321 agcccgaccc tcccctggca caacgtaagg ccattatctc tcatccaact ccaggacgga
    10381 gtcagtgaga tcgatcaaat gcgcctcgac actagttcga accatagagc ccaccgcatc
    10441 cccagcatgc ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc
    10501 ccccagaata gaatgacacc tactcagaca atgcgatgca atttcctcat tttattagga
    10561 aaggacagtg ggagtggcac cttccagggt caaggaaggc acgggggagg ggcaaacaac
    10621 agatggctgg caactagaag gcacagtcga ggcactagtt tacccgggga gcatgtcaag
    10681 gtcaaaatcg tcaagagcgt cagcaggcag catatcaagg tcaaagtcgt caagggcatc
    10741 ggctgggagc atgtctaagt caaaatcgtc aagggcgtcg gccggcccgc cgctttcgca
    10801 ctttagctgt ttctccaggc cacatatgat tagttccagg ccgaaaagga aggcaggttc
    10861 ggctccctgc cggtcgaaca gctcaattgc ttgtctcaga agtgggggca tagaatcggt
    10921 ggtaggtgtc tctctttcct cttttgctac ttgatgctcc tgttcctcca atacgcagcc
    10981 cagtgtaaag tggcccacgg cggacagagc gtacagtgcg ttctccaggg agaagccttg
    11041 ctgacacagg aacgcgagct gattttccag ggtttcgtac tgtttctctg ttgggcgggt
    11101 gccgagatgc actttagccc cgtcgcgatg tgagaggaga gcacagcgga atgacttggc
    11161 gttgttccgc agaaagtctt gccatgactc gccttccagg gggcagaagt gggtatgatg
    11221 cctgtccagc atctcgattg gcagggcatc gagcagggcc cgcttgttct tcacgtgcca
    11281 gtacagggta ggctgctcaa ctcccagctt ttgagcgagt ttccttgtcg tcaggccttc
    11341 gataccgact ccattgagta attccagagc gccgtttatg actttgctct tgtccagtct
    11401 agacatgaat tctttgccaa aatgatgaga cagcacaata accagcacgt tgcccaggag
    11461 ctgtaggaaa aagaagaagg catgaacatg gttagcagag gctctagagc cgccggtcac
    11521 acgccagaag ccgaaccccg ccctgccccg tcccccccga aggcagccgt ccccccgcgg
    11581 acagccccga ggctggagag ggagaagggg acggcggcgc ggcgacgcac gaaggccctc
    11641 cccgcccatt tccttcctgc cggcgccgca ccgcttcgcc ccgcgcccgc tagagggggt
    11701 gcggcggcgc ctcccagatt tcggctccgc acagatttgg gacaaaggaa gtccctgcgc
    11761 cctctcgcac gattaccata aaaggcaatg gctgcggctc gccgcgcctc gacagccgcc
    11821 ggcgctccgg gggccgccgc gcccctcccc cgagccctcc ccggcccgag gcggccccgc
    11881 cccgcccggc acccccacct gccgccaccc cccgcccggc acggcgagcc ccgcgccacg
    11941 ccccgtacgg agccccgcac ccgaagccgg gccgtgctca gcaactcggg gaggggggtg
    12001 cagggggggt tgcagcccga ccgacgcgcc cacaccccct gctcaccccc ccacgcacac
    12061 accccgcacg cagcctttgt tcccctcgca gccccccccg caccgcgggg caccgccccc
    12121 ggccgcgctc ccctcgcgca cactgcggag cgcacaaagc cccgcgccgc gcccgcagcg
    12181 ctcacagccg ccgggcagcg cggagccgca cgcggcgctc cccacgcaca cacacacgca
    12241 cgcacccccc gagccgctcc ccccgcacaa agggccctcc cggagcccct caaggctttc
    12301 acgcagccac agaaaagaaa caagccgtca ttaaaccaag cgctaattac agcccggagg
    12361 agaagggccg tcccgcccgc tcacctgtgg gagtaacgcg gtcagtcaga gccggggcgg
    12421 gcggcgcgag gcggcggcgg agcggggcac ggggcgaagg cagcgcgcag cgactcccgc
    12481 ccgccgcgcg cttcgctttt tatagggccg ccgccgccgc cgcctcgcca taaaaggaaa
    12541 ctttcggagc gcgccgctct gattggctgc cgccgcacct ctccgcctcg ccccgccccg
    12601 cccctcgccc cgccccgccc cgcctggcgc gcgccccccc cccccccccg cccccatcgc
    12661 tgcacaaaat aattaaaaaa taaataaata caaaattggg ggtggggagg ggggggagat
    12721 ggggagagtg aagcagaacg tggggctcac ctcgaccatc atggtaatag cgatgactaa
    12781 tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca
    12841 ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg
    12901 atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt
    12961 ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt
    13021 gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg
    13081 gctatgaact aatgaccccg taattgatta ctattaataa ctagctagtc aataatcaat
    13141 gtcgacgatg atgctgtaca ctcgaaactt ttttcgaagg tccgaggttc tagacgagtt
    13201 tactccctat cagtgataga gaacgatgtc gagtttac
//
