LOCUS       sequence-217842-       13265 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 13265)
  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 13265)
  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..13265
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    659..1462
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     protein_bind    1469..1502
                     /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    1506..1531
                     /label=SA
                     /note="splice acceptor site"
     CDS             1552..1605
                     /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"
     CDS             1615..2214
                     /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(1615..1634)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2111..2131
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    2368..2479
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_signal    4083..4130
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4155..4174
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4194..5624)
                     /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"
     regulatory      5621..5630
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      5714..5723
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(5731..5755)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(5752..5790)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    5802..5820
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5838..5856
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5873..5891
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5909..5927
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5945..5963
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5980..5998
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     enhancer        6058..6441
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6446..6723
                     /label=chicken beta-actin promoter
     intron          6724..7740
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     7664..7686
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     7748..7767
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(7823..8440,8441..8446,8447..8482,8483..8485,8486..8521,
                     8522..8524,8525..8560,8561..8569)
                     /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"
     polyA_signal    8583..8807
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    10168..10201
                     /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)"
     misc_feature    10211..11047
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     primer_bind     complement(11077..11097)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(11077..11095)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(11116..11132)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(11116..11132)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(11129..11151)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    11140..11156
                     /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)."
     promoter        complement(join(11164..11170,11171..11188,11189..11194))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11209..11230
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(11347..11364)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(11518..12106)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(11598..11617)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(12277..13068,13069..13137))
                     /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     12900..12919
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(13138..13242)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtacctg
      661 ctttctctga ccagcattct ctcccctggg cctgtgccgc tttctgtctg cagcttgtgg
      721 cctgggtcac ctctacggct ggcccagatc cttccctgcc gcctccttca ggttccgtct
      781 tcctccactc cctcttcccc ttgctctctg ctgtgttgct gcccaaggat gctctttccg
      841 gagcacttcc ttctcggcgc tgcaccacgt gatgtcctct gagcggatcc tccccgtgtc
      901 tgggtcctct ccgggcatct ctcctccctc acccaacccc atgccgtctt cactcgctgg
      961 gttccctttt ccttctcctt ctggggcctg tgccatctct cgtttcttag gatggccttc
     1021 tccgacggat gtctcccttg cgtcccgcct ccccttcttg taggcctgca tcatcaccgt
     1081 ttttctggac aaccccaaag taccccgtct ccctggcttt agccacctct ccatcctctt
     1141 gctttctttg cctggacacc ccgttctcct gtggattcgg gtcacctctc actcctttca
     1201 tttgggcagc tcccctaccc cccttacctc tctagtctgt gctagctctt ccagccccct
     1261 gtcatggcat cttccagggg tccgagagct cagctagtct tcttcctcca acccgggccc
     1321 ctatgtccac ttcaggacag catgtttgct gcctccaggg atcctgtgtc cccgagctgg
     1381 gaccacctta tattcccagg gccggttaat gtggctctgg ttctgggtac ttttatctgt
     1441 cccctccacc ccacagtggg gcaagcttga agttcctatt cttcaaatag tataggaact
     1501 tcggcctgac ctcttctctt cctcccacag ggcctcagat ctggcagcgg agagggcaga
     1561 ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctaggct cgaaatgacc
     1621 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc
     1681 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac
     1741 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     1801 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     1861 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     1921 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     1981 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     2041 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     2101 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     2161 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgacgcccg
     2221 ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatggctc cgaccgaagc
     2281 cacccggggc ggccccgccg accccgcacc cgcccccgag gcccaccgac tctagagggc
     2341 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     2401 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     2461 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     2521 tggggcagga cagcaagggg gaggattggg aagacaatag cactcgacac tagtaagcgt
     2581 atcgatacgg ggacagcccc cccccaaagc ccccagggat gtaattacgt ccctcccccg
     2641 ctagggggca gcagcgagcc gcccggggct ccgctccggt ccggcgctcc ccccgcatcc
     2701 ccgagccggc agcgtgcggg gacagcccgg gcacggggaa ggtggcacgg gatcgctttc
     2761 ctctgaacgc ttctcgctgc tctttgagcc tgcagacacc tggggggata cggggaaaaa
     2821 gctttaggct tgtgtctgag cctgcatgtt tgatggtgtc tggatgcaag cagaaggggt
     2881 ggaagagctt gcctggagag atacagctgg gtcagtagga ctgggacagg cagctggaga
     2941 attgccatgt agatgttcat acaatcgtca aatcatgaag gctggaaaag ccctccaaga
     3001 tccccaagac caaccccaac ccacccaccg tgcccactgg ccatgtccct cagtgccaca
     3061 tccccacagt tcttcatcac ctccagggac ggtgaccccc ccacctccgt gggcagctgt
     3121 gccactgcag caccgctctt tggagaaggt aaatcttgct aaatccagcc cgaccctccc
     3181 ctggcacaac gtaaggccat tatctctcat ccaactccag gacggagtca gtgagatgca
     3241 tacggggaca gccccccccc aaagccccca gggatgtaat tacgtccctc ccccgctagg
     3301 gggcagcagc gagccgcccg gggctccgct ccggtccggc gctccccccg catccccgag
     3361 ccggcagcgt gcggggacag cccgggcacg gggaaggtgg cacgggatcg ctttcctctg
     3421 aacgcttctc gctgctcttt gagcctgcag acacctgggg ggatacgggg aaaaagcttt
     3481 aggcttgtgt ctgagcctgc atgtttgatg gtgtctggat gcaagcagaa ggggtggaag
     3541 agcttgcctg gagagataca gctgggtcag taggactggg acaggcagct ggagaattgc
     3601 catgtagatg ttcatacaat cgtcaaatca tgaaggctgg aaaagccctc caagatcccc
     3661 aagaccaacc ccaacccacc caccgtgccc actggccatg tccctcagtg ccacatcccc
     3721 acagttcttc atcacctcca gggacggtga cccccccacc tccgtgggca gctgtgccac
     3781 tgcagcaccg ctctttggag aaggtaaatc ttgctaaatc cagcccgacc ctcccctggc
     3841 acaacgtaag gccattatct ctcatccaac tccaggacgg agtcagtgag atcgatcaaa
     3901 tgcgcttcga tctcgagcta tggcagggcc tgccgccccg acgttggctg cgagccctgg
     3961 gccttcaccc gaacttgggg ggtggggtgg ggaaaaggaa gaaacgcggg cgtattggcc
     4021 ccaatggggt ctcggtgggg tatcgacaga gtgccagccc tgggaccgaa ccccgcgttt
     4081 atgaacaaac gacccaacac cgtgcgtttt attctgtctt tttattgccg tcatagcgcg
     4141 ggttccttcc ggtattgtct ccttccgtgt ttcagttagc ctcccccacg cgtttacttg
     4201 tacagctcgt ccatgccgta caggaacagg tggtggcggc cctcggagcg ctcgtactgt
     4261 tccacgatgg tgtagtcctc gttgtgggag gtgatgtcca gcttggtgtc cacgtagtag
     4321 tagccgggca gttgcacggg cttcttggcc atgtagatgg tcttgaactc caccaggtag
     4381 tggccgccgt ccttcagctt cagggcctgg tggatctcgc ccttcagcac gccgtcgcgg
     4441 gggtacaggc gctcggtgga ggcctcccag cccatggtct tcttctgcat tacggggccg
     4501 tcggggggga agttggtgcc gcgcatcttc accttgtaga tcagcgtgcc gtcctgcagg
     4561 gaggagtcct gggtcacggt caccagaccg ccgtcctcga agttcatcac gcgctcccac
     4621 ttgaagccct cggggaagga cagcttcttg taatcgggga tgtcggcggg gtgcttcacg
     4681 tacgccttgg agccgtacat gaactggggg gacaggatgt cccaggcgaa gggcaggggg
     4741 ccgcccttgg tcaccttcag cttggcggtc tgggtgccct cgtaggggcg gccctcgccc
     4801 tcgccctcga tctcgaactc gtggccgttc atggagccct ccatgcgcac cttgaagcgc
     4861 atgaactctt tgatgacggc catgttgttg tcctcggagg aggcggtgcc ggagctgccg
     4921 ctgccggtgc tgccggtgcc atgccccagg aacaggtggt ggcggccctc ggagcgctcg
     4981 tactgttcca cgatggtgta gtcctcgttg tgggaggtga tgtccagctt ggtgtccacg
     5041 tagtagtagc cgggcagttg cacgggcttc ttggccatgt agatggtctt gaactccacc
     5101 aggtagtggc cgccgtcctt cagcttcagg gcctggtgga tctcgccctt cagcacgccg
     5161 tcgcgggggt acaggcgctc ggtggaggcc tcccagccca tggtcttctt ctgcattacg
     5221 gggccgtcgg gggggaagtt ggtgccgcgc atcttcacct tgtagatcag cgtgccgtcc
     5281 tgcagggagg agtcctgggt cacggtcacc agaccgccgt cctcgaagtt catcacgcgc
     5341 tcccacttga agccctcggg gaaggacagc ttcttgtaat cggggatgtc ggcggggtgc
     5401 ttcacgtacg ccttggagcc gtacatgaac tggggggaca ggatgtccca ggcgaagggc
     5461 agggggccgc ccttggtcac cttcagcttg gcggtctggg tgccctcgta ggggcggccc
     5521 tcgccctcgc cctcgatctc gaactcgtgg ccgttcatgg agccctccat gcgcaccttg
     5581 aagcgcatga actctttgat gacctcctcg cccttgctca ccatggtggc atatggatat
     5641 ctccaccggt cgtcatcgtc atccttgtaa tcaatatcat gatctttata atcaccgtca
     5701 tggtctttgt agtccatggt ggccttaagc gatctgacgg ttcactaaac gagctctgct
     5761 tatataggcc tcccaccgta cacgcctacc tcgacatacg ttctctatca ctgataggga
     5821 gtaaactcga catacgttct ctatcactga tagggataaa ctcgacatac gttctctatc
     5881 actgataggg agtaaactcg acatacgttc tctatcactg atagggagta aactcgacat
     5941 acgttctcta tcactgatag ggagtaaact cgacatcgtt ctctatcact gatagggagt
     6001 aaactcgtct agaacctcgg accttcgaaa aagtttcgag tgtacagcat catcgtcgac
     6061 attgattatt gactagctag ttattaatag taatcaatta cggggtcatt agttcatagc
     6121 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     6181 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     6241 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     6301 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     6361 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     6421 ttagtcatcg ctattaccat gatggtcgag gtgagcccca cgttctgctt cactctcccc
     6481 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
     6541 gcgatggggg cggggggggg gggggggcgc gcgccaggcg gggcggggcg gggcgagggg
     6601 cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag
     6661 tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg
     6721 gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc
     6781 ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct
     6841 cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt
     6901 gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg
     6961 tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag
     7021 cgctgcgggc gcggcgcggg gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc
     7081 gggggcggtg ccccgcggtg cggggggggc tgcgagggga acaaaggctg cgtgcggggt
     7141 gtgtgcgtgg gggggtgagc agggggtgtg ggcgcgtcgg tcgggctgca accccccctg
     7201 cacccccctc cccgagttgc tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg
     7261 cgtggcgcgg ggctcgccgt gccgggcggg gggtggcggc aggtgggggt gccgggcggg
     7321 gcggggccgc ctcgggccgg ggagggctcg ggggaggggc gcggcggccc ccggagcgcc
     7381 ggcggctgtc gaggcgcggc gagccgcagc cattgccttt tatggtaatc gtgcgagagg
     7441 gcgcagggac ttcctttgtc ccaaatctgt gcggagccga aatctgggag gcgccgccgc
     7501 accccctcta gcgggcgcgg ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg
     7561 gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc ctctccagcc tcggggctgt
     7621 ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt
     7681 gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag
     7741 ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcatttaa
     7801 atatgcatac gtataccaat tgatgtctag actggacaag agcaaagtca taaacggcgc
     7861 tctggaatta ctcaatggag tcggtatcga aggcctgacg acaaggaaac tcgctcaaaa
     7921 gctgggagtt gagcagccta ccctgtactg gcacgtgaag aacaagcggg ccctgctcga
     7981 tgccctgcca atcgagatgc tggacaggca tcatacccac ttctgccccc tggaaggcga
     8041 gtcatggcaa gactttctgc ggaacaacgc caagtcattc cgctgtgctc tcctctcaca
     8101 tcgcgacggg gctaaagtgc atctcggcac ccgcccaaca gagaaacagt acgaaaccct
     8161 ggaaaatcag ctcgcgttcc tgtgtcagca aggcttctcc ctggagaacg cactgtacgc
     8221 tctgtccgcc gtgggccact ttacactggg ctgcgtattg gaggaacagg agcatcaagt
     8281 agcaaaagag gaaagagaga cacctaccac cgattctatg cccccacttc tgagacaagc
     8341 aattgagctg ttcgaccggc agggagccga acctgccttc cttttcggcc tggaactaat
     8401 catatgtggc ctggagaaac agctaaagtg cgaaagcggc gggccggccg acgcccttga
     8461 cgattttgac ttagacatgc tcccagccga tgcccttgac gactttgacc ttgatatgct
     8521 gcctgctgac gctcttgacg attttgacct tgacatgctc cccgggtaaa ctagtgcctc
     8581 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     8641 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     8701 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     8761 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggttc gaactagtgt
     8821 cgaggcgcat ttgatcgatc tcactgactc cgtcctggag ttggatgaga gataatggcc
     8881 ttacgttgtg ccaggggagg gtcgggctgg atttagcaag atttaccttc tccaaagagc
     8941 ggtgctgcag tggcacagct gcccacggag gtgggggggt caccgtccct ggaggtgatg
     9001 aagaactgtg gggatgtggc actgagggac atggccagtg ggcacggtgg gtgggttggg
     9061 gttggtcttg gggatcttgg agggcttttc cagccttcat gatttgacga ttgtatgaac
     9121 atctacatgg caattctcca gctgcctgtc ccagtcctac tgacccagct gtatctctcc
     9181 aggcaagctc ttccacccct tctgcttgca tccagacacc atcaaacatg caggctcaga
     9241 cacaagccta aagctttttc cccgtatccc cccaggtgtc tgcaggctca aagagcagcg
     9301 agaagcgttc agaggaaagc gatcccgtgc caccttcccc gtgcccgggc tgtccccgca
     9361 cgctgccggc tcggggatgc ggggggagcg ccggaccgga gcggagcccc gggcggctcg
     9421 ctgctgcccc ctagcggggg agggacgtaa ttacatccct gggggctttg ggggggggct
     9481 gtccccgtat gcatctcact gactccgtcc tggagttgga tgagagataa tggccttacg
     9541 ttgtgccagg ggagggtcgg gctggattta gcaagattta ccttctccaa agagcggtgc
     9601 tgcagtggca cagctgccca cggaggtggg ggggtcaccg tccctggagg tgatgaagaa
     9661 ctgtggggat gtggcactga gggacatggc cagtgggcac ggtgggtggg ttggggttgg
     9721 tcttggggat cttggagggc ttttccagcc ttcatgattt gacgattgta tgaacatcta
     9781 catggcaatt ctccagctgc ctgtcccagt cctactgacc cagctgtatc tctccaggca
     9841 agctcttcca ccccttctgc ttgcatccag acaccatcaa acatgcaggc tcagacacaa
     9901 gcctaaagct ttttccccgt atccccccag gtgtctgcag gctcaaagag cagcgagaag
     9961 cgttcagagg aaagcgatcc cgtgccacct tccccgtgcc cgggctgtcc ccgcacgctg
    10021 ccggctcggg gatgcggggg gagcgccgga ccggagcgga gccccgggcg gctcgctgct
    10081 gccccctagc gggggaggga cgtaattaca tccctggggg ctttgggggg gggctgtccc
    10141 cgtatcgata cgcttgtcga gatcgaagaa gttcctattc tctagaaagt ataggaactt
    10201 cgcggccgct actagggaca ggattggtga cagaaaagcc ccatccttag gcctcctcct
    10261 tcctagtctc ctgatattgg gtctaacccc cacctcctgt taggcagatt ccttatctgg
    10321 tgacacaccc ccatttcctg gagccatctc tctccttgcc agaacctcta aggtttgctt
    10381 acgatggagc cagagaggat cctgggaggg agagcttggc agggggtggg agggaagggg
    10441 gggatgcgtg acctgcccgg ttctcagtgg ccaccctgcg ctaccctctc ccagaacctg
    10501 agctgctctg acgcggctgt ctggtgcgtt tcactgatcc tggtgctgca gcttccttac
    10561 acttcccaag aggagaagca gtttggaaaa acaaaatcag aataagttgg tcctgagttc
    10621 taactttggc tcttcacctt tctagtcccc aatttatatt gttcctccgt gcgtcagttt
    10681 tacctgtgag ataaggccag tagccagccc cgtcctggca gggctgtggt gaggaggggg
    10741 gtgtccgtgt ggaaaactcc ctttgtgaga atggtgcgtc ctaggtgttc accaggtcgt
    10801 ggccgcctct actccctttc tctttctcca tccttctttc cttaaagagt ccccagtgct
    10861 atctgggaca tattcctccg cccagagcag ggtcccgctt ccctaaggcc ctgctctggg
    10921 cttctgggtt tgagtccttg gcaagcccag gagaggcgct caggcttccc tgtccccctt
    10981 cctcgtccac catctcatgc ccctggctct cctgcccctt ccctacaggg gttcctggct
    11041 ctgctctcca ccgcggtgga gctccagctt ttgttccctt tagtgagggt taattgcgcg
    11101 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
    11161 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
    11221 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
    11281 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
    11341 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
    11401 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
    11461 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
    11521 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
    11581 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
    11641 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
    11701 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
    11761 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
    11821 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
    11881 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
    11941 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
    12001 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
    12061 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
    12121 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
    12181 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
    12241 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
    12301 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
    12361 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
    12421 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
    12481 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
    12541 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
    12601 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
    12661 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
    12721 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
    12781 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
    12841 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
    12901 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
    12961 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
    13021 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
    13081 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
    13141 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
    13201 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
    13261 gccac
//
