LOCUS       sequence-217840-       10292 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 
            constitutive expression of your protein of interest..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10292)
  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 10292)
  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:F3-P CAGGS tdTPH-F
FEATURES             Location/Qualifiers
     source          1..10292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(129..197,198..989)
                     /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(347..366)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1160..1748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1649..1668
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1902..1919
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2036..2057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2072..2077,2078..2095,2096..2102)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2110..2126
                     /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     2115..2137
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2134..2150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2134..2150
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2169..2189
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2171..2189
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    2219..3055
                     /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(3065..3098)
                     /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)"
     enhancer        3128..3511
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3516..3793
                     /label=chicken beta-actin promoter
     intron          3794..4810
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     4734..4756
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     4818..4837
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      4904..4913
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4910..6337
                     /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"
     CDS             6359..6415
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             6428..7453
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             6428..7453
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    7583..7704
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7620..7639)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7674..7693
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     polyA_signal    7814..7925
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     7914..7931
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(8079..8678)
                     /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(8162..8182)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     8659..8678
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(8688..8741)
                     /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    8762..8787
                     /label=SA
                     /note="splice acceptor site"
     protein_bind    complement(8791..8824)
                     /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    8831..9634
                     /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(9648..9667)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(9649..9667)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(9677..9694)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(9677..9693)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9686..9708)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      9835..10290
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(9922..9941)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     10132..10153
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
       61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
      541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
      601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
      661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
      721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
      781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
      841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
      901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
      961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     1801 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     1861 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     1921 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     1981 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     2041 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     2101 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     2161 ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctccac cgcggtggag
     2221 agcagagcca ggaacccctg tagggaaggg gcaggagagc caggggcatg agatggtgga
     2281 cgaggaaggg ggacagggaa gcctgagcgc ctctcctggg cttgccaagg actcaaaccc
     2341 agaagcccag agcagggcct tagggaagcg ggaccctgct ctgggcggag gaatatgtcc
     2401 cagatagcac tggggactct ttaaggaaag aaggatggag aaagagaaag ggagtagagg
     2461 cggccacgac ctggtgaaca cctaggacgc accattctca caaagggagt tttccacacg
     2521 gacacccccc tcctcaccac agccctgcca ggacggggct ggctactggc cttatctcac
     2581 aggtaaaact gacgcacgga ggaacaatat aaattgggga ctagaaaggt gaagagccaa
     2641 agttagaact caggaccaac ttattctgat tttgtttttc caaactgctt ctcctcttgg
     2701 gaagtgtaag gaagctgcag caccaggatc agtgaaacgc accagacagc cgcgtcagag
     2761 cagctcaggt tctgggagag ggtagcgcag ggtggccact gagaaccggg caggtcacgc
     2821 atccccccct tccctcccac cccctgccaa gctctccctc ccaggatcct ctctggctcc
     2881 atcgtaagca aaccttagag gttctggcaa ggagagagat ggctccagga aatgggggtg
     2941 tgtcaccaga taaggaatct gcctaacagg aggtgggggt tagacccaat atcaggagac
     3001 taggaaggag gaggcctaag gatggggctt ttctgtcacc aatcctgtcc ctagtagcgg
     3061 ccgcgaagtt cctatacttt ctagagaata ggaacttctt cgatctcgag tgtacagcat
     3121 catcgtcgac attgattatt gactagctag ttattaatag taatcaatta cggggtcatt
     3181 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     3241 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     3301 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     3361 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     3421 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     3481 catctacgta ttagtcatcg ctattaccat gatggtcgag gtgagcccca cgttctgctt
     3541 cactctcccc atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt
     3601 attttgtgca gcgatggggg cggggggggg gggggggcgc gcgccaggcg gggcggggcg
     3661 gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc
     3721 gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag
     3781 cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc
     3841 ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga
     3901 cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct
     3961 gtggctgcgt gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc
     4021 ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg
     4081 cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg
     4141 gagcgcggcc gggggcggtg ccccgcggtg cggggggggc tgcgagggga acaaaggctg
     4201 cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca
     4261 accccccctg cacccccctc cccgagttgc tgagcacggc ccggcttcgg gtgcggggct
     4321 ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc aggtgggggt
     4381 gccgggcggg gcggggccgc ctcgggccgg ggagggctcg ggggaggggc gcggcggccc
     4441 ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc
     4501 gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag
     4561 gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga
     4621 aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc
     4681 tcggggctgt ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg
     4741 cttctggcgt gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt
     4801 tttcctacag ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga
     4861 attcatttaa atatgcatac gtataccaat tgatatcacc ggtgccacca tggtgagcaa
     4921 gggcgaggag gtcatcaaag agttcatgcg cttcaaggtg cgcatggagg gctccatgaa
     4981 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
     5041 cgccaagctg aaggtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc
     5101 ccagttcatg tacggctcca aggcgtacgt gaagcacccc gccgacatcc ccgattacaa
     5161 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
     5221 tctggtgacc gtgacccagg actcctccct gcaggacggc acgctgatct acaaggtgaa
     5281 gatgcgcggc accaacttcc cccccgacgg ccccgtaatg cagaagaaga ccatgggctg
     5341 ggaggcctcc accgagcgcc tgtacccccg cgacggcgtg ctgaagggcg agatccacca
     5401 ggccctgaag ctgaaggacg gcggccacta cctggtggag ttcaagacca tctacatggc
     5461 caagaagccc gtgcaactgc ccggctacta ctacgtggac accaagctgg acatcacctc
     5521 ccacaacgag gactacacca tcgtggaaca gtacgagcgc tccgagggcc gccaccacct
     5581 gttcctgggg catggcaccg gcagcaccgg cagcggcagc tccggcaccg cctcctccga
     5641 ggacaacaac atggccgtca tcaaagagtt catgcgcttc aaggtgcgca tggagggctc
     5701 catgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac
     5761 ccagaccgcc aagctgaagg tgaccaaggg cggccccctg cccttcgcct gggacatcct
     5821 gtccccccag ttcatgtacg gctccaaggc gtacgtgaag caccccgccg acatccccga
     5881 ttacaagaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga
     5941 cggcggtctg gtgaccgtga cccaggactc ctccctgcag gacggcacgc tgatctacaa
     6001 ggtgaagatg cgcggcacca acttcccccc cgacggcccc gtaatgcaga agaagaccat
     6061 gggctgggag gcctccaccg agcgcctgta cccccgcgac ggcgtgctga agggcgagat
     6121 ccaccaggcc ctgaagctga aggacggcgg ccactacctg gtggagttca agaccatcta
     6181 catggccaag aagcccgtgc aactgcccgg ctactactac gtggacacca agctggacat
     6241 cacctcccac aacgaggact acaccatcgt ggaacagtac gagcgctccg agggccgcca
     6301 ccacctgttc ctgtacggca tggacgagct gtacaagggt ggtggctcgg ggggaggagc
     6361 gacaaacttt agcttgctga agcaagctgg tgacgttgag gagaatcccg gaccaggatt
     6421 cacgcgtatg aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa
     6481 gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag
     6541 cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta
     6601 caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc cggaagtgct
     6661 tgacattggg gaattcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt
     6721 cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc
     6781 catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc cattcggacc
     6841 gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca
     6901 tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct
     6961 cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga
     7021 tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag
     7081 cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg
     7141 gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg
     7201 atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt
     7261 ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg
     7321 atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac
     7381 cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag
     7441 ggcaaaggaa tagcacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt
     7501 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     7561 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     7621 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     7681 gtccaaactc atcaatgtat cttatcatgt ctgttcgaac tagtgtcgag tgctattgtc
     7741 ttcccaatcc tcccccttgc tgtcctgccc caccccaccc cccagaatag aatgacacct
     7801 actcagacaa tgcgatgcaa tttcctcatt ttattaggaa aggacagtgg gagtggcacc
     7861 ttccagggtc aaggaaggca cgggggaggg gcaaacaaca gatggctggc aactagaagg
     7921 cacagtcgag gctgatcagc gggtttaaac gggccctcta gagtcggtgg gcctcggggg
     7981 cgggtgcggg gtcggcgggg ccgccccggg tggcttcggt cggagccatg gggtcgtgcg
     8041 ctcctttcgg tcgggcgctg cgggtcgtgg ggcgggcgtc aggcaccggg cttgcgggtc
     8101 atgcaccagg tgcgcggtcc ttcgggcacc tcgacgtcgg cggtgacggt gaagccgagc
     8161 cgctcgtaga aggggaggtt gcggggcgcg gaggtctcca ggaaggcggg caccccggcg
     8221 cgctcggccg cctccactcc ggggagcacg acggcgctgc ccagaccctt gccctggtgg
     8281 tcgggcgaga cgccgacggt ggccaggaac cacgcgggct ccttgggccg gtgcggcgcc
     8341 aggaggcctt ccatctgttg ctgcgcggcc agccgggaac cgctcaactc ggccatgcgc
     8401 gggccgatct cggcgaacac cgcccccgct tcgacgctct ccggcgtggt ccagaccgcc
     8461 accgcggcgc cgtcgtccgc gacccacacc ttgccgatgt cgagcccgac gcgcgtgagg
     8521 aagagttctt gcagctcggt gacccgctcg atgtggcggt ccgggtcgac ggtgtggcgc
     8581 gtggcggggt agtcggcgaa cgcggcggcg agggtgcgta cggcccgggg gacgtcgtcg
     8641 cgggtggcga ggcgcaccgt gggcttgtac tcggtcattt cgagcctagg gccgggattc
     8701 tcctccacgt caccgcatgt tagaagactt cctctgccct ctccgctgcc agatctgagg
     8761 ccctgtggga ggaagagaag aggtcaggcc gaagttccta tactatttga agaataggaa
     8821 cttcaagctt gccccactgt ggggtggagg ggacagataa aagtacccag aaccagagcc
     8881 acattaaccg gccctgggaa tataaggtgg tcccagctcg gggacacagg atccctggag
     8941 gcagcaaaca tgctgtcctg aagtggacat aggggcccgg gttggaggaa gaagactagc
     9001 tgagctctcg gacccctgga agatgccatg acagggggct ggaagagcta gcacagacta
     9061 gagaggtaag gggggtaggg gagctgccca aatgaaagga gtgagaggtg acccgaatcc
     9121 acaggagaac ggggtgtcca ggcaaagaaa gcaagaggat ggagaggtgg ctaaagccag
     9181 ggagacgggg tactttgggg ttgtccagaa aaacggtgat gatgcaggcc tacaagaagg
     9241 ggaggcggga cgcaagggag acatccgtcg gagaaggcca tcctaagaaa cgagagatgg
     9301 cacaggcccc agaaggagaa ggaaaaggga acccagcgag tgaagacggc atggggttgg
     9361 gtgagggagg agagatgccc ggagaggacc cagacacggg gaggatccgc tcagaggaca
     9421 tcacgtggtg cagcgccgag aaggaagtgc tccggaaaga gcatccttgg gcagcaacac
     9481 agcagagagc aaggggaaga gggagtggag gaagacggaa cctgaaggag gcggcaggga
     9541 aggatctggg ccagccgtag aggtgaccca ggccacaagc tgcagacaga aagcggcaca
     9601 ggcccagggg agagaatgct ggtcagagaa agcaggtacc caattcgccc tatagtgagt
     9661 cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     9721 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     9781 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc
     9841 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     9901 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     9961 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
    10021 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
    10081 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
    10141 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
    10201 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
    10261 attttaacaa aatattaacg cttacaattt ag
//
