LOCUS       sequence-217839-        9824 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genome engineering donor vector for creating master cell lines 
            suitable for FLPe recombinase-mediated cassette exchange (RMCE) in 
            the AAVS1 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9824)
  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 9824)
  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-CAGGS GPHTK-F
FEATURES             Location/Qualifiers
     source          1..9824
                     /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"
     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    1513..1546
                     /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)"
     enhancer        1552..1935
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1940..2217
                     /label=chicken beta-actin promoter
     intron          2218..3234
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3158..3180
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3242..3261
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      3295..3304
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3301..3303,3304..3306,3307..4017)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3301..3303,3304..3306,3307..4017)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3346..3367)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3607..3626)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             4039..4095
                     /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             5188..6183
                     /codon_start=1
                     /product="truncated herpes simplex virus thymidine kinase 
                     (Salomon et al., 1995)"
                     /label=Delta-TK
                     /note="confers sensitivity to nucleoside analogs such as 
                     acyclovir or ganciclovir"
                     /translation="MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWRV
                     LGASETIANIYTTQHRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGS
                     SHAPPPALTLIFDRHPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGAL
                     PEDRHIDRLAKRQRPGERLELAMLAAIRRVYGLLANTVRYLQCGGSWREDWGQLSGTAV
                     PPQGAEPQSNAGPRPHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRSMHVFI
                     LDYDQSPAGCRDALLQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     primer_bind     complement(6181..6200)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6225..6273
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    6669..6702
                     /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)"
     promoter        complement(6710..6728)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    6770..7606
                     /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(7636..7656)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7636..7654)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7675..7691)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7675..7691)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7688..7710)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7699..7715
                     /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(7723..7729,7730..7747,7748..7753))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7768..7789
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7906..7923)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8077..8665)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8157..8176)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8836..9627,9628..9696))
                     /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     9459..9478
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9697..9801)
                     /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 attctctaga aatattctcg aggtttaaac
     1501 gtcgacacta gtgaagttcc tattcttcaa atagtatagg aacttcggcg cgccattgat
     1561 tattgactag ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     1621 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     1681 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     1741 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     1801 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     1861 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     1921 atcgctatta ccatgatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc
     1981 cccccctccc cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg
     2041 ggggcggggg gggggggggg gcgcgcgcca ggcggggcgg ggcggggcga ggggcggggc
     2101 ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct
     2161 tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga
     2221 gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc
     2281 cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg
     2341 ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc
     2401 cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg
     2461 tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc
     2521 gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc
     2581 ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc
     2641 gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc
     2701 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc
     2761 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     2821 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
     2881 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     2941 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
     3001 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     3061 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
     3121 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     3181 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     3241 ggcaacgtgc tggttattgt gctgtctcat cattttggca aacaattgga tgctgccacc
     3301 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     3361 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     3421 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     3481 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     3541 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     3601 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     3661 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     3721 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     3781 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     3841 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     3901 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     3961 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggt
     4021 ggtggctcgg ggggaggagc gacaaacttt agcttgctga agcaagctgg tgacgttgag
     4081 gagaatcccg gaccaggatt cacgcgtatg aaaaagcctg aactcaccgc gacgtctgtc
     4141 gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc
     4201 gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat
     4261 agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg
     4321 ctcccgattc cggaagtgct tgacattggg gaattcagcg agagcctgac ctattgcatc
     4381 tcccgccgtg cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt
     4441 ctgcagccgg tcgcggaggc catggatgcg atcgctgcgg ccgatcttag ccagacgagc
     4501 gggttcggcc cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata
     4561 tgcgcgattg ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt
     4621 gcgtccgtcg cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc
     4681 cggcacctcg tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata
     4741 acagcggtca ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac
     4801 atcttcttct ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg
     4861 aggcatccgg agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt
     4921 gaccaactct atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt
     4981 cgatgcgacg caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc
     5041 agaagcgcgg ccgtctggac cgatggctgt gtagaagtcg cgtctgcgtt cgaccaggct
     5101 gcgcgttctc gcggccatag caaccgacgt acggcgttgc gccctcgccg gcagcaagaa
     5161 gccacggaag tccgcccgga gcagaaaatg cccacgctac tgcgggttta tatagacggt
     5221 ccccacggga tggggaaaac caccaccacg caactgctgg tggccctggg ttcgcgcgac
     5281 gatatcgtct acgtacccga gccgatgact tactggcggg tgctgggggc ttccgagaca
     5341 atcgcgaaca tctacaccac acaacaccgc ctcgaccagg gtgagatatc ggccggggac
     5401 gcggcggtgg taatgacaag cgcccagata acaatgggca tgccttatgc cgtgaccgac
     5461 gccgttctgg ctcctcatat cgggggggag gctgggagct cacatgcccc gcccccggcc
     5521 ctcaccctca tcttcgaccg ccatcccatc gccgccctcc tgtgctaccc ggccgcgcgg
     5581 taccttatgg gcagcatgac cccccaggcc gtgctggcgt tcgtggccct catcccgccg
     5641 accttgcccg gcaccaacat cgtgcttggg gcccttccgg aggacagaca catcgaccgc
     5701 ctggccaaac gccagcgccc cggcgagcgg ctggaactgg ctatgctggc tgcgattcgc
     5761 cgcgtttacg ggctacttgc caatacggtg cggtatctgc agtgcggcgg gtcgtggcgg
     5821 gaggactggg gacagctttc ggggacggcc gtgccgcccc agggtgccga gccccagagc
     5881 aacgcgggcc cacgacccca tatcggggac acgttattta ccctgtttcg ggcccccgag
     5941 ttgctggccc ccaacggcga cctgtataac gtgtttgcct gggccttgga cgtcttggcc
     6001 aaacgcctcc gttccatgca cgtctttatc ctggattacg accaatcgcc cgccggctgc
     6061 cgggacgccc tgctgcaact tacctccggg atggtccaga cccacgtcac cacccccggc
     6121 tccataccga cgatatgcga cctggcgcgc acgtttgccc gggagatggg ggaggctaac
     6181 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     6241 gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     6301 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     6361 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     6421 cggcaggccc tgccatagcc actggccccg tgggttaggg acggggtccc ccatggggaa
     6481 tggtttatgg ttcgtggggg ttattatttt gggcgttgcg tggggtcagg tccacgaccc
     6541 aagcttgcgg gactctgggg ttcgaataaa gaccgaccaa gcgacgtctg agagctccct
     6601 ggcgaattcg gtaccaataa aagagcttta ttttcatgat ctgtgtgttg gtttttgtgt
     6661 gcggcgcgga agttcctatt ctctagaaag tataggaact tcctcgagcc ctatagtgag
     6721 tcgtattaaa gggcgaattc tgcagatatc catcacactg gcggccgcta ctagggacag
     6781 gattggtgac agaaaagccc catccttagg cctcctcctt cctagtctcc tgatattggg
     6841 tctaaccccc acctcctgtt aggcagattc cttatctggt gacacacccc catttcctgg
     6901 agccatctct ctccttgcca gaacctctaa ggtttgctta cgatggagcc agagaggatc
     6961 ctgggaggga gagcttggca gggggtggga gggaaggggg ggatgcgtga cctgcccggt
     7021 tctcagtggc caccctgcgc taccctctcc cagaacctga gctgctctga cgcggctgtc
     7081 tggtgcgttt cactgatcct ggtgctgcag cttccttaca cttcccaaga ggagaagcag
     7141 tttggaaaaa caaaatcaga ataagttggt cctgagttct aactttggct cttcaccttt
     7201 ctagtcccca atttatattg ttcctccgtg cgtcagtttt acctgtgaga taaggccagt
     7261 agccagcccc gtcctggcag ggctgtggtg aggagggggg tgtccgtgtg gaaaactccc
     7321 tttgtgagaa tggtgcgtcc taggtgttca ccaggtcgtg gccgcctcta ctccctttct
     7381 ctttctccat ccttctttcc ttaaagagtc cccagtgcta tctgggacat attcctccgc
     7441 ccagagcagg gtcccgcttc cctaaggccc tgctctgggc ttctgggttt gagtccttgg
     7501 caagcccagg agaggcgctc aggcttccct gtcccccttc ctcgtccacc atctcatgcc
     7561 cctggctctc ctgccccttc cctacagggg ttcctggctc tgctctccac cgcggtggag
     7621 ctccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
     7681 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     7741 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     7801 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     7861 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     7921 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     7981 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     8041 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     8101 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     8161 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     8221 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     8281 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     8341 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     8401 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     8461 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     8521 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     8581 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     8641 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     8701 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     8761 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     8821 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     8881 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     8941 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     9001 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     9061 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     9121 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     9181 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     9241 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     9301 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     9361 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     9421 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     9481 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     9541 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     9601 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     9661 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     9721 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     9781 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccac
//
