LOCUS       sequence-229049-        9824 bp    DNA     circular SYN 14-APR-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 Apr 14, 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"
     promoter        309..586
                     /label=chicken beta-actin promoter
     intron          587..1603
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1527..1549
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1611..1630
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1664..1673
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1670..1672,1673..1675,1676..2386)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1670..1672,1673..1675,1676..2386)
                     /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(1715..1736)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1976..1995)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2408..2464
                     /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             3557..4552
                     /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(4550..4569)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4594..4642
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     protein_bind    5038..5071
                     /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(5079..5097)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    5139..5975
                     /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(6005..6025)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6005..6023)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6044..6060)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6044..6060)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6057..6079)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6068..6084
                     /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(6092..6098,6099..6116,6117..6122))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6137..6158
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6275..6292)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6446..7034)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6526..6545)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7205..7996,7997..8065))
                     /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     7828..7847
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8066..8170)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8196..8651)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8333..8354)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8545..8564
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8778..8800
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8792..8809
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8793..8809
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8819..8837
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    8852..9655
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     enhancer        join(9745..9824,1..304)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc
      361 acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg
      421 gggggggggg sgcgcgccag gcggggcggg gcggggcgag gggcggggcg gggcgaggcg
      481 gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag
      541 gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg
      601 ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact
      661 gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta
      721 gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct
      781 ccgggagggc cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt gtgtgtgcgt
      841 ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc
      901 ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg
      961 gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg tgggggggtg
     1021 agcagggggt gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc ctccccgagt
     1081 tgctgagcac ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg cggggctcgc
     1141 cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc
     1201 cggggagggc tcgggggagg ggcgcggcgg cccccggagc gccggcggct gtcgaggcgc
     1261 ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg gacttccttt
     1321 gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct ctagcgggcg
     1381 cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc ttcgtgcgtc
     1441 gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg gggacggctg
     1501 ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag
     1561 agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct
     1621 ggttattgtg ctgtctcatc attttggcaa acaattggat gctgccacca tggtgagcaa
     1681 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     1741 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     1801 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     1861 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     1921 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     1981 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     2041 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     2101 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     2161 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     2221 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     2281 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     2341 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagggtg gtggctcggg
     2401 gggaggagcg acaaacttta gcttgctgaa gcaagctggt gacgttgagg agaatcccgg
     2461 accaggattc acgcgtatga aaaagcctga actcaccgcg acgtctgtcg agaagtttct
     2521 gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg
     2581 tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga
     2641 tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc
     2701 ggaagtgctt gacattgggg aattcagcga gagcctgacc tattgcatct cccgccgtgc
     2761 acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt
     2821 cgcggaggcc atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc
     2881 attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc
     2941 tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc
     3001 gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt
     3061 gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat
     3121 tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg
     3181 gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga
     3241 gcttgcagga tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta
     3301 tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc
     3361 aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc
     3421 cgtctggacc gatggctgtg tagaagtcgc gtctgcgttc gaccaggctg cgcgttctcg
     3481 cggccatagc aaccgacgta cggcgttgcg ccctcgccgg cagcaagaag ccacggaagt
     3541 ccgcccggag cagaaaatgc ccacgctact gcgggtttat atagacggtc cccacgggat
     3601 ggggaaaacc accaccacgc aactgctggt ggccctgggt tcgcgcgacg atatcgtcta
     3661 cgtacccgag ccgatgactt actggcgggt gctgggggct tccgagacaa tcgcgaacat
     3721 ctacaccaca caacaccgcc tcgaccaggg tgagatatcg gccggggacg cggcggtggt
     3781 aatgacaagc gcccagataa caatgggcat gccttatgcc gtgaccgacg ccgttctggc
     3841 tcctcatatc gggggggagg ctgggagctc acatgccccg cccccggccc tcaccctcat
     3901 cttcgaccgc catcccatcg ccgccctcct gtgctacccg gccgcgcggt accttatggg
     3961 cagcatgacc ccccaggccg tgctggcgtt cgtggccctc atcccgccga ccttgcccgg
     4021 caccaacatc gtgcttgggg cccttccgga ggacagacac atcgaccgcc tggccaaacg
     4081 ccagcgcccc ggcgagcggc tggaactggc tatgctggct gcgattcgcc gcgtttacgg
     4141 gctacttgcc aatacggtgc ggtatctgca gtgcggcggg tcgtggcggg aggactgggg
     4201 acagctttcg gggacggccg tgccgcccca gggtgccgag ccccagagca acgcgggccc
     4261 acgaccccat atcggggaca cgttatttac cctgtttcgg gcccccgagt tgctggcccc
     4321 caacggcgac ctgtataacg tgtttgcctg ggccttggac gtcttggcca aacgcctccg
     4381 ttccatgcac gtctttatcc tggattacga ccaatcgccc gccggctgcc gggacgccct
     4441 gctgcaactt acctccggga tggtccagac ccacgtcacc acccccggct ccataccgac
     4501 gatatgcgac ctggcgcgca cgtttgcccg ggagatgggg gaggctaact gaaacacgga
     4561 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     4621 acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     4681 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     4741 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     4801 gccatagcca ctggccccgt gggttaggga cggggtcccc catggggaat ggtttatggt
     4861 tcgtgggggt tattattttg ggcgttgcgt ggggtcaggt ccacgaccca agcttgcggg
     4921 actctggggt tcgaataaag accgaccaag cgacgtctga gagctccctg gcgaattcgg
     4981 taccaataaa agagctttat tttcatgatc tgtgtgttgg tttttgtgtg cggcgcggaa
     5041 gttcctattc tctagaaagt ataggaactt cctcgagccc tatagtgagt cgtattaaag
     5101 ggcgaattct gcagatatcc atcacactgg cggccgctac tagggacagg attggtgaca
     5161 gaaaagcccc atccttaggc ctcctccttc ctagtctcct gatattgggt ctaaccccca
     5221 cctcctgtta ggcagattcc ttatctggtg acacaccccc atttcctgga gccatctctc
     5281 tccttgccag aacctctaag gtttgcttac gatggagcca gagaggatcc tgggagggag
     5341 agcttggcag ggggtgggag ggaagggggg gatgcgtgac ctgcccggtt ctcagtggcc
     5401 accctgcgct accctctccc agaacctgag ctgctctgac gcggctgtct ggtgcgtttc
     5461 actgatcctg gtgctgcagc ttccttacac ttcccaagag gagaagcagt ttggaaaaac
     5521 aaaatcagaa taagttggtc ctgagttcta actttggctc ttcacctttc tagtccccaa
     5581 tttatattgt tcctccgtgc gtcagtttta cctgtgagat aaggccagta gccagccccg
     5641 tcctggcagg gctgtggtga ggaggggggt gtccgtgtgg aaaactccct ttgtgagaat
     5701 ggtgcgtcct aggtgttcac caggtcgtgg ccgcctctac tccctttctc tttctccatc
     5761 cttctttcct taaagagtcc ccagtgctat ctgggacata ttcctccgcc cagagcaggg
     5821 tcccgcttcc ctaaggccct gctctgggct tctgggtttg agtccttggc aagcccagga
     5881 gaggcgctca ggcttccctg tcccccttcc tcgtccacca tctcatgccc ctggctctcc
     5941 tgccccttcc ctacaggggt tcctggctct gctctccacc gcggtggagc tccagctttt
     6001 gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg
     6061 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     6121 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     6181 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     6241 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     6301 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     6361 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     6421 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6481 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6541 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6601 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6661 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6721 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6781 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6841 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6901 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6961 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     7021 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     7081 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     7141 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     7201 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     7261 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     7321 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     7381 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     7441 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     7501 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     7561 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7621 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7681 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7741 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7801 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7861 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7921 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7981 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     8041 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     8101 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     8161 gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt
     8221 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     8281 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     8341 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     8401 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     8461 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     8521 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     8581 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     8641 acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     8701 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     8761 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta
     8821 atacgactca ctatagggcg aattgggtac ctgctttctc tgaccagcat tctctcccct
     8881 gggcctgtgc cgctttctgt ctgcagcttg tggcctgggt cacctctacg gctggcccag
     8941 atccttccct gccgcctcct tcaggttccg tcttcctcca ctccctcttc cccttgctct
     9001 ctgctgtgtt gctgcccaag gatgctcttt ccggagcact tccttctcgg cgctgcacca
     9061 cgtgatgtcc tctgagcgga tcctccccgt gtctgggtcc tctccgggca tctctcctcc
     9121 ctcacccaac cccatgccgt cttcactcgc tgggttccct tttccttctc cttctggggc
     9181 ctgtgccatc tctcgtttct taggatggcc ttctccgacg gatgtctccc ttgcgtcccg
     9241 cctccccttc ttgtaggcct gcatcatcac cgtttttctg gacaacccca aagtaccccg
     9301 tctccctggc tttagccacc tctccatcct cttgctttct ttgcctggac accccgttct
     9361 cctgtggatt cgggtcacct ctcactcctt tcatttgggc agctccccta ccccccttac
     9421 ctctctagtc tgtgctagct cttccagccc cctgtcatgg catcttccag gggtccgaga
     9481 gctcagctag tcttcttcct ccaacccggg cccctatgtc cacttcagga cagcatgttt
     9541 gctgcctcca gggatcctgt gtccccgagc tgggaccacc ttatattccc agggccggtt
     9601 aatgtggctc tggttctggg tacttttatc tgtcccctcc accccacagt ggggcaagct
     9661 tgaattctct agaaatattc tcgaggttta aacgtcgaca ctagtgaagt tcctattctt
     9721 caaatagtat aggaacttcg gcgcgccatt gattattgac tagctagtta ttaatagtaa
     9781 tcaattacgg ggtcattagt tcatagccca tatatggagt tccg
//
